Home-based supervised exercise versus hospital-based supervised exercise or unsupervised walk advice as treatment for intermittent claudication: a systematic review.
To evaluate the effects of home-based supervised exercise vs hospital-based supervised exercise, and the effects of home-based supervised exercise vs unsupervised "go home and walk advice" on daily life and corridor-walking capacity, health-related quality of life and patient-reported functional walking capacity in patients with intermittent claudication. Systematic literature searches were conducted in PubMed, EMBASE, ProQuest, the Cumulative Index to Nursing and Allied Health Literature (CINAHL), the Allied and Complementary Medicine Database (AMED), the Cochrane Library, and a number of Health Technology Assessment (HTA)-databases in October 2014. Randomized controlled trials and non-randomized controlled trials (> 100 patients) were considered for inclusion. Data extraction and risk of bias assessment was performed independently and discussed in meetings. Seven randomized controlled trials and 2 non-randomized controlled studies fulfilled the inclusion criteria. The included studies had some, or major, limitations. Based on a low quality of evidence, home-based supervised exercise may lead to less improvement in maximum and pain-free walking distance, and in more improvement in daily life walking capacity, compared with hospital-based supervised exercise. Home-based supervised exercise may improve maximum and pain-free walking distance compared with "go home and walk advice" and result in little or no difference in health-related quality of life and functional walking capacity compared with hospital-based supervised exercise or "go home and walk advice". Further research is needed to establish the optimal exercise modality for these patients.


[Comparative characteristics of microbial proteases by the level of hydrolysis of protein substrates].
Screening of enzyme preparations displaying a maximum proteolytic activity at pH 4.0-5.5 and effecting deep proteolysis of plant proteins was performed. Amyloprotooryzin prepared from Aspergillus oryzae 387 containing a complex of proteolytic enzymes was the most effective. The amino acid composition of the hydrolysates obtained was studied. Amyloprotooryzin increased the contents of amino acids by 108-227%, depending on the substrate used. The enzymatic complex of amyloprotooryzin was studied; in addition, proteases, alpha-amylase, exo-beta-glucanase, and xylanase were detected in the complex.


Individuals with multiple sclerosis redistribute positive mechanical work from the ankle to the hip during walking.
Individuals with multiple sclerosis (MS) typically walk slower, have reduced cadences and shorter step lengths. While these spatiotemporal gait alterations have been thought to be due to decreased power generation at the ankle, the distribution of mechanical work across the ankle, knee and hip joints during walking is not well understood. By quantifying the mechanical work at each joint, the compensatory mechanisms utilized by individuals with MS to maintain gait speed may be better understood. Fifteen subjects with MS (EDSS: 4.4±1.0) and fifteen healthy age-matched control subjects completed a three-dimensional gait analysis. The net mechanical work at the ankle, knee and hip joints was quantified for the stance phase of gait. Our results found that the less impaired leg of the subjects with MS generated a similar amount of mechanical work as the control group; however, the ankle joint produced less positive mechanical work, and the hip joint generated more positive mechanical work. Additionally, the less impaired leg of the subjects with MS and the leg of the control group generated more positive work than the more impaired leg of the subjects with MS. These outcomes suggest that individuals with MS may adopt a hip compensatory strategy with their less impaired leg during gait due to the limited amount of mechanical work generated at the ankle.


Pituitary adenylate cyclase-activating polypeptide (PACAP) protects against mitoxantrone-induced cardiac injury in mice.
Mitoxantrone (MXT) is an androstenedione that is used to treat cancers and progressive forms of multiple sclerosis; however, its use is limited by its cardiotoxicity. Pituitary adenylate cyclase activating polypeptide (PACAP) is a member of the secretin/growth hormone-releasing hormone/vasoactive intestinal peptide family and has many functions, including cytoprotection and immunosuppression. We tested the hypothesis that PACAP can protect against MXT-induced cardiotoxicity in mice. Female BALB/c mice were treated once weekly for 4 weeks with saline (n=14) or MXT (3mg/kg, i.p.; n=14). Half of the mice in each group received PACAP (10μg, i.p.) 1h before and 24 and 48h after MXT, while the remaining mice received injections of saline on the same schedule. Echocardiography was used to assess cardiac structure and function. In mice treated with MXT and saline, body weight was significantly reduced after the third dose of MXT. PACAP significantly attenuated the reduction in body weight; however, the weights did not return to control level. Compared to controls, MXT-treated mice had significantly increased left ventricular (LV) diameter and LV volume and decreased LV posterior wall thickness. Fractional shortening (FS) and ejection fraction (EF) were also significantly decreased. Treatment with PACAP prevented MXT-induced LV dilation and significantly attenuated the reductions in FS and EF, although FS and EF did not return to control level. PACAP38 did not prevent MXT-induced decreases in LV posterior wall thickness. MXT dose-dependently decreased the viability of cultured U937 (human leukemia) cells; PACAP did not protect cultured U937 cells from MXT-mediated cell death. In conclusion, PACAP can attenuate MXT-mediated LV dilation and dysfunction in mice.


Rapid and Efficient Separation of Decursin and Decursinol Angelate from Angelica gigas Nakai using Ionic Liquid, (BMIm)BF4, Combined with Crystallization.
Ionic liquids (ILs) have gained much attention as alternative solvents to volatile organic solvents due to their attractive properties. This study aimed to develop an efficient method for the selective separation of decursin (D) and decursinol angelate (DA) from Angelica gigas Nakai (A. gigas) using ILs and crystallization. The IL 1-butyl-3-methylimidazolium tetrafluoroborate ((BMIm)BF4) was the most efficient at extracting D and DA. Parameters including solid-to-liquid ratio, time, and temperature were optimized by response surface methodology (RSM). Under optimal extraction conditions (1 g/6.5 mL solid-to-liquid ratio, 60 °C temperature, and 120 min time), the extraction yields of D and DA were 43.32 mg/g (97.06%) and 17.87 mg/g (97.12%), respectively. Moreover, drowning out crystallization using deionized water (DW) as an anti-solvent offered an excellent ability to recover D and DA from the A. gigas-(BMIm)BF4 extraction solution. The rates of recovery and the total purity of D and DA were found to be greater than 97%. Therefore, a rapid and efficient method of combining ILs with crystallization was effectively achieved for the selective separation of D and DA. This approach is assumed to be beneficial in the pharmaceutical industry for the effective obtention of D- and DA-enriched products.


Antiretroviral Treatment for HIV Elite Controllers?
In most HIV-infected persons, the natural history of untreated infection is one of sustained viremia, progressive CD4 T cell depletion with resultant morbidity and mortality. The advent of effective combination antiretroviral therapy (ART) that controls HIV replication has altered this landscape dramatically. Yet a rare population of HIV-infected persons-elite controllers (EC)-can control HIV replication such that plasma levels of virus are "undetectable" without ART. The EC phenotype is heterogeneous, with some subjects durably controlling the virus-persistent elite controllers-and some eventually losing viral control-transient elite controllers. Overall, EC tend to have robust HIV-specific T cell responses and in some cases, mainly in transient elite controllers, elevated activation and inflammation indices that diminish with ART suggesting that endogenous defenses against this persistent pathogen come at the cost of heightened activation/inflammation. A limited data set suggests that cardiovascular disease risk as well as the occur-rence of other morbid events may be greater in the overall EC population than in treated HIV infection. ART in EC decreases activation indices but does not appear to increase circulating CD4 T cell numbers nor do we know if it alters clinical outcomes. Thus, it is difficult to recommend or discourage a decision to start ART in the EC population but the authors lean toward treatment particularly in those EC whose activation indices are high and those who are progressively losing circulating CD4 T cell numbers. Biomarkers that can reliably predict loss of virologic control and immune failure are needed.


SPATIAL AND TEMPORAL VARIATION IN SELECTION ON CORRELATED LIFE-HISTORY TRAITS AND PLANT SIZE IN CHAMAECRISTA FASCICULATA.
Intraspecific studies of selection on multiple traits of a plant's life history provide insight as to how the composite life history of an organism evolves. Current understanding of selection on plant life-history traits is deficient in three important areas: 1) the effects of selection through correlated traits, 2) the effects of selection on a trait throughout the plant's lifetime, and 3) spatial and temporal variation in selection on plant life-history traits among populations and years. This study documents spatial and temporal variation in selection on three life-history and two morphological traits for two natural populations of Chamaecrista fasciculata, a native summer annual. Life-history and morphological traits (date of seedling emergence, size at establishment, size prior to reproduction, date of initial flowering, and date of initial fruit maturation) varied significantly between sites and/or years. Selection on traits varied either spatially, between sites and among transects within one site, or temporally, between years. In addition, life-history traits were phenotypically correlated among themselves and with morphological traits; correlations were generally constant over time and space. Indirect selection caused changes in means and variances in traits not under direct selection, but which were correlated with traits under selection. Selection on date of emergence varied in direction and magnitude among different life-cycle stages, while selection on other traits varied only in magnitude among life stages of the plant. This study documents the complexity of the selective process and the importance of considering multiple life stages and traits when studying the evolution of life-history traits.


Photoactive Metal-Organic Framework and Its Film for Light-Driven Hydrogen Production and Carbon Dioxide Reduction.
The design of a new photocatalytic system and integrating the essential components in a structurally controlled manner to create artificially photosynthetic systems is high desirable. By incorporating a photoactive triphenylamine moiety to assemble a Gd-based metal-organic framework as a heterogeneous photosensitizer, new artificial systems were constructed for the proton and carbon dioxide reduction under irradiation. The assembled MOFs exhibited a one-dimensional metal-oxygen pillar that was connected together by the depronated TCA(3-) ligands to form a three-dimensional noninterpenetrating porous framework. The combining of proton reduction and/or the carbon dioxide reduction catalysts, i.e., the Fe-Fe hydrogenase active site models and the Ni(Cyclam) complexes, initiated a photoinduced single electron transfer from its excited state to the substrate. The system exhibited an initial TOF of 320 h(-1) of hydrogen per catalyst and an overall quantum yield of about 0.21% and is able to reduce carbon dioxide under irradiation. The deposit of the photoactive Gd-TCA into the film of an α-Al2O3 plate provided a platform for the practical applications through prolonging the lifetime of the artifical system and allowed the easily operated devices being recyclable as a promising photocatalytic system.


Exploiting Pharmacological Similarity to Identify Safety Concerns - Listen to What the Data Tells You.
Whilst most new drugs are designed to act on a single target or a small number of targets, many do show broad pharmacological activity. In some cases this can be beneficial and necessary for efficacy and in others it can be detrimental, leading to increased safety liability. To probe off-target pharmacology most drug discovery programs include screening against a broad panel of targets that represent known troublesome pharmacology. Hits against any one of these targets can then be subjected to a risk assessment for potential safety problems in preclinical or clinical studies. In addition, the secondary pharmacology profile can also be thought of as an alternative description of the compound and as such can be used as a method for assessing 'similarity'. Consequently, inspection of the in vivo findings of pharmacological neighbors can give important insights into potential safety liabilities that are neither identified by pure chemical similarity searches nor by risk assessment on individual targets. Here we show that the pharmacological profile contains additional information as compared to chemical similarity, and also demonstrate how this can be used in the hazard assessment done during drug discovery and development.


The enduring impact of historical and structural racism on urban violence in Philadelphia.
Public health approaches to crime and injury prevention are increasingly focused on the physical places and environments where violence is concentrated. In this study, our aim is to explore the association between historic place-based racial discrimination captured in the 1937 Home Owners Loan Corporation (HOLC) map of Philadelphia and present-day violent crime and firearm injuries. The creators of the 1937 HOLC map zoned Philadelphia based in a hierarchical system wherein first-grade and green color zones were used to indicate areas desirable for government-backed mortgage lending and economic development, a second-grade or blue zone for areas that were already developed and stable, a third-grade or yellow zone for areas with evidence of decline and influx of a "low grade population," and fourth-grade or red zone for areas with dilapidated or informal housing and an "undesirable population" of predominately Black residents. We conducted an empirical spatial analysis of the concentration of firearm assaults and violent crimes in 2013 through 2014 relative to zoning in the 1937 HOLC map. After adjusting for socio-demographic factors at the time the map was created from the 1940 Census, firearm injury rates are highest in historically red-zoned areas of Philadelphia. The relationship between HOLC map zones and general violent crime is not supported after adjusting for historical Census data. This analysis extends historic perspective to the relationship between emplaced structural racism and violence, and situates the socio-ecological context in which people live at the forefront of this association.


Coordination of family's care in an evidence-based nurse home visiting program.
Interprofessional care coordination within evidence-based prevention programs like Nurse-Family Partnership® (NFP) is necessary to meet family needs and maximize program impact. This study aimed to describe the coordination of families' care in the NFP home visiting context. We used an adapted grounded theory approach and purposively sampled seven NFP sites. We conducted telephone interviews with 95 participants: 51 NFP staff (54%), 39 healthcare providers (41%), and 5 social service providers (5%). All interviews were recorded, transcribed, validated, and analyzed in NVivo11. Many community providers in all sites described their knowledge of the characteristics of the NFP intervention, including the strength of its evidence to achieve outcomes. Care coordination was dynamic and changed over time based on client needs and staff willingness to work together. Effective care coordination in the NFP context from the provider perspective is driven by shared knowledge, integrated systems, mission alignment, and individual champions who value the program.


Interactional influence of fatigue and task difficulty on cardiovascular response: demonstrations involving an aerobic exercise challenge.
Experiments were executed to provide new evidence relevant to the recent suggestion that fatigue should augment or retard cardiovascular response depending on the difficulty of the challenge at hand. Participants walked on a treadmill while wearing a vest fitted with 5 or 25 pounds of weight. Later, they mounted a recumbent stationary bicycle and were asked to pedal with the chance to earn a modest incentive if they attained a low or high cycling standard (i.e., if they met an easy or difficult cycling challenge). Analysis of CV responses during the cycling period indicated expected interactions for systolic blood pressure and heart rate. Whereas responses were stronger for the Heavy-Vest (i.e., high-fatigue) group when the standard was low, they were weaker for this group when the standard was high. Experiments 2 and 3 evaluated a nonfatigue interpretation of the main results and yielded findings that supported the fatigue interpretation.


Decoding the biosynthesis and function of diphthamide, an enigmatic modification of translation elongation factor 2 (EF2)
Diphthamide is a highly conserved modification of archaeal and eukaryal translation elongation factor 2 (EF2) and yet why cells need EF2 to contain diphthamide is unclear. In yeast, the first steps of diphthamide synthesis and the genes (DPH1-DPH5) required to form the intermediate diphthine are well-documented. However, the last step, amidation of diphthine to diphthamide, had largely been ill-defined. Remarkably, through mining genome-wide synthetic gene array (SGA) and chemical genomics databases, recent studies by Uthman et al. [PLoS Genetics (2013) 9, e1003334] and Su et al. [Proc. Natl. Acad. Sci. USA (2012) 109, 19983-19987] have identified two more diphthamide players, DPH6 and DPH7. Consistent with roles in the amidation step, dph6 and dph7 deletion strains fail to complete diphthamide synthesis and accumulate diphthine-modified EF2. In contrast to Dph6, the catalytically relevant amidase, Dph7 appears to be regulatory. As shown by Uthman et al., it promotes dissociation of diphthine synthase (Dph5) from EF2, allowing diphthine amidation by Dph6 to occur and thereby coupling diphthine synthesis to the terminal step in the pathway. Remarkably, the study by Uthman et al. suggests that Dph5 has a novel role as an EF2 inhibitor that affects cell growth when diphthamide synthesis is blocked or incomplete and, importantly, shows that diphthamide promotes the accuracy of EF2 performance during translation.


PEAKS: powerful software for peptide de novo sequencing by tandem mass spectrometry.
A number of different approaches have been described to identify proteins from tandem mass spectrometry (MS/MS) data. The most common approaches rely on the available databases to match experimental MS/MS data. These methods suffer from several drawbacks and cannot be used for the identification of proteins from unknown genomes. In this communication, we describe a new de novo sequencing software package, PEAKS, to extract amino acid sequence information without the use of databases. PEAKS uses a new model and a new algorithm to efficiently compute the best peptide sequences whose fragment ions can best interpret the peaks in the MS/MS spectrum. The output of the software gives amino acid sequences with confidence scores for the entire sequences, as well as an additional novel positional scoring scheme for portions of the sequences. The performance of PEAKS is compared with Lutefisk, a well-known de novo sequencing software, using quadrupole-time-of-flight (Q-TOF) data obtained for several tryptic peptides from standard proteins.


Intractable childhood epilepsy and maternal fatigue.
Mothers of children with intractable epilepsy are generally stressed and experience more emotional problems. Fatigue may affect their productivity, social interactions, and their ability to adequately take care of their children. The objectives were to examine the relationship between intractable childhood epilepsy and maternal fatigue, and explore possible contributing factors. Sixty-four consecutive mothers of children with intractable epilepsy were identified prospectively. Exclusion criteria included degenerative/metabolic disorders or life threatening illness, such as brain tumors. Fatigue was measured using a standardized 11-item questionnaire, which has been revalidated in an Arabic speaking population. Mothers' ages were 24-45 years (mean 34) and ages of their epileptic children were 1-15 years (mean 6.7). Most children (64%) had epilepsy for >2 years, were on >1 antiepileptic drug (AED) (72%), and had daily seizures (47%). Thirty-four (54%) of the children had motor deficits and 83% had mental retardation (severe in 41%). Twenty-eight (44%) mothers were fatigued. Factors associated with increased maternal fatigue included child's age <2 years (p=0.01), cryptogenic epilepsy (p=0.03), and severe motor deficits (p=0.04). Factors associated with lowered fatigue included performing regular exercise (p=0.006), lack of mental retardation (p=0.01), seizure control (p=0.05), using one AED (p=0.002), infrequent ER visits (p=0.005), and lack of recent hospitalization (p=0.005). Mothers of children with intractable epilepsy are increasingly fatigued. Several correlating factors were identified, mostly related to seizure control, mental and physical handicap. Strategies to manage the problem include proper education, seizure control, participation in regular exercise, social support, and psychological counseling.


Molar pregnancy and living normal fetus coexisting until term: prenatal biochemical and sonographic diagnosis.
The extremely rare condition of molar pregnancy with a coexisting fetus progressing to a viable infant, is reported. At 20 weeks gestational age, prenatal diagnosis was made by biochemical and sonographic findings, in which elevated free beta-human chorionic gonadotrophin (HCG, 500 ng/ml) was noted and ultrasound showed a well-defined multicystic snowstorm-like mass connecting with placenta. Chromosomal evaluation by amniocentesis was normal (46,XY) and ultrasound showed no fetal abnormalities, so the patient decided to keep the pregnancy and had no significant complications noted in the antepartal period. At 38 weeks gestational age, a Caesarean section was performed due to cephalopelvic disproportion and a 3380 g, living boy was delivered. The infant did not show any abnormality. The placenta and the connecting hydatidiform molar tissue were delivered manually. Subsequently, since persistent elevated beta-HCG was noted 2 months later, the patient was treated with chemotherapy for gestational trophoblastic disease. A normal beta-HCG concentration was noted thereafter. Prenatal differential diagnosis and postnatal management are discussed as well as the rationale for allowing the pregnancy to continue after the diagnosis at 20 weeks gestational age.


Access and Quality of HIV-Related Point-of-Care Diagnostic Testing in Global Health Programs.
Access to point-of-care testing (POCT) improves patient care, especially in resource-limited settings where laboratory infrastructure is poor and the bulk of the population lives in rural settings. However, because of challenges in rolling out the technology and weak quality assurance measures, the promise of human immunodeficiency virus (HIV)-related POCT in resource-limited settings has not been fully exploited to improve patient care and impact public health. Because of these challenges, the Joint United Nations Programme on HIV/AIDS (UNAIDS), in partnership with other organizations, recently launched the Diagnostics Access Initiative. Expanding HIV programs, including the "test and treat" strategies and the newly established UNAIDS 90-90-90 targets, will require increased access to reliable and accurate POCT results. In this review, we examine various components that could improve access and uptake of quality-assured POC tests to ensure coverage and public health impact. These components include evaluation, policy, regulation, and innovative approaches to strengthen the quality of POCT.


Experience Using Kaolin-Impregnated Sponge to Minimize Perioperative Bleeding in Norwood Operation.
A kaolin-impregnated hemostatic sponge (QuikClot) is reported to reduce intraoperative blood loss in trauma and noncardiac surgery. The purpose of this study was to assess if this sponge was effective for hemostasis during Norwood operation. We conducted a retrospective review of patients undergoing Norwood operation in infancy between 2011 and 2016 at our institution. Of 31 identified Norwood operations, a kaolin-impregnated sponge was used intraoperatively in 15 (48%) patients. The preoperative profiles and cardiopulmonary bypass status were similar between the operations with or without kaolin-impregnated sponge. The comparison on each operative outcome between operations with or without kaolin-impregnated sponge showed that the intraoperative platelets, cryoprecipitate, and factor VII dosage were significantly less in the operations with kaolin-impregnated sponge (55 mL, 10 mL, 0 µg/kg vs 72 mL, 15 mL, 45 µg/kg; P = .03, .021, .019), as well as the incidence of perioperative bleeding complications (second cardiopulmonary bypass for hemostasis or postoperative mediastinal exploration, 0% vs 31%, P = .043). A logistic regression model showed that the nonuse of kaolin-impregnated sponge and longer aortic cross clamp time were associated with perioperative bleeding complication in univariable model ( P = .02 and .005). Use of kaolin-impregnated hemostatic sponge was associated with reduced blood product use and perioperative bleeding complications in Norwood operation at a single institution.


Interpretation Modalities Used on Family-Centered Rounds: Perspectives of Spanish-Speaking Families.
Limited-English-proficient (LEP) patients and families are at risk for worse hospital outcomes due to impaired communication. Family-centered rounds (FCR) have become the preferred format for information sharing among providers and families at pediatric institutions. However, there are limited data on FCR among LEP families, particularly regarding interpretation type. We sought to examine the relationships between interpretation type and FCR satisfaction and efficacy among Spanish-speaking families, the fastest growing LEP population in pediatric hospitals. Spanish speakers admitted to general pediatrics units over a period of 16 months were identified on admission. A bilingual research assistant observed FCR and completed an observation tool, including interpreter type. After FCR, the research assistant interviewed families, collecting parent demographics, FCR experience, and interpreter satisfaction. Associations between interpretation type and satisfaction as well as interpretation type and understanding were tested by using χ2 analyses. We studied 124 families over 16 months. Most respondents were patients' mothers (84%), born in Mexico (76%), had grade school education (56%), and spoke limited English (96%). Overall, 83 (73%) reported the interpreter services improved their understanding of their child's medical condition. Interpreter type was significantly associated with family satisfaction with FCR; specifically, an in-person hospital interpreter or video interpreter was associated with complete caregiver satisfaction (P = .005). Spanish speakers report higher satisfaction with face-to-face interpreters during FCR, including in-person and video, compared with telephonic interpreters. Video interpretation via iPad during FCR may be a valuable and accessible approach to improve communication in the care of hospitalized children.


Game-based training to promote handwashing, handrub and gloving for hospital pharmacy operators.
Adherence to handwashing, handrub and gloving procedures is mandatory for safe, aseptic drug compounding in hospital pharmacies. This study measured participants' satisfaction and effectiveness of a game-based training tool (Handtastic Box) developed to improve adherence. Handtastic Boxes were played by pairs of pharmacy operators (introductory video, 1 min study of guidelines, game). In module 1, players watched videos of somebody handwashing and had to find the missing step. They examined wooden models of hands under ultraviolet (UV) light, with some areas stained with fluorescein, to find the hand showing contamination. In module 2, players used a fluorescein hydroalcoholic solution and placed their hands under UV light to highlight missing areas. In module 3, players identified major errors that could compromise glove sterility and linked them to a problem explanation. Then, they applied paint to their fingertips and donned gloves-the paint had to stay inside them. Satisfaction about the training was assessed with a 10-question survey; knowledge about procedures was assessed using a before-and-after questionnaire of nine questions, a 100-point confidence score (modules 1 and 2), and the number of before-and-after errors made during donning gloves (module 3). Operators were very satisfied and felt more competent after training. Average knowledge score increased from 56.3% (SD 18.2%) to 93.7% (SD 9.5%), and confidence in answers increased from 66.4% (SD 18.7%) to 95.7% (SD 5.52%) (n=14, both modules 1 and 2). The mean error score for gloving procedure decreased from 1.7 (SD 0.8%) to 0.3 (SD 0.5%) (n=10, module 3). Handtastic Boxes proved to be a highly effective training method for improving knowledge of handwashing, handrub and gloving.


Rotation and diffusion of H2 in hydrogen-ice clathrate by 1H NMR.
A recently reported hydrogen-ice clathrate carries up to four H(2) in each large cage and one H(2) in each small cage. We report pulsed proton NMR line shape measurements on H(2)-D(2)O clathrate formed at 1500 bar and 250 K. The behavior of the two-pulse spin-echo amplitude with respect to the nutation angle of the refocusing pulse shows that intramolecular dipolar broadening, modulated by H(2) molecular reorientations, dominates the line width of the ortho-H(2). Dipolar interaction between H(2) guests and host D atoms explains the echo variation with the relative phases of the pulses. From 12 to 120 K, the line width varies as 1/T, demonstrating that the three sublevels of J = 1 are split by a constant energy, epsilon. The splitting arises from distortion in the otherwise high-symmetry cages from frozen-out D(2)O orientational disorder. Above 120 K, further line-narrowing signals the onset of H(2) diffusion from cage to cage. At the lowest temperature, 1.9 K, the spectrum has Pake powder doublet-like features; the doublet is not fully developed, indicating a broad distribution of order parameters and energies epsilon.


Incidence of venous thromboembolism in rheumatoid arthritis, results from a "real-life" cohort and an appraisal of available literature.
Rheumatoid arthritis (RA) is associated with an increased risk of venous thromboembolism (VTE) occurrence. In this work, we assessed the incidence and predictive factors of VTE in our "real-life" cohort of RA patients. To contextualize our results, we reviewed the available literature about this topic.We performed a retrospective analysis of prospectively followed-up patients with RA attending our Rheumatologic Clinic between January 2010 and December 2020. Each patient was investigated for VTE occurrence. Incident cases were reported as incidence proportion and incidence rate per 1000 person-years at risk. Possible predictive factors were also exploited by regression analyses. Available literature about this topic was also assessed.In this evaluation, 347 consecutive patients without previous evidence of VTE, attending our Rheumatologic Clinic from 2010 to 2020, were studied. In our "real-life" cohort, the incidence proportion of VTE was 3.7% (2.7-4.7%) and considering over 1654 person-years, an incidence rate of 7.8 × 1000 (2.5-11.7). Exploratively assessing predictive factors in our cohort, older age (hazard ratio [HR] 1.07, 95% confidence interval [CI] 1.01-1.14, p = .015), higher body mass index (HR 1.37, 95% CI 1.04-1.80, P = .026), and longer disease duration (HR 1.11, 95% CI 1.03-1.20, P = .006) resulted to be significant predictors of VTE occurrence during the follow-up.In our "real-life" cohort, VTE burden has been suggested in patients with RA. Comparing our results with previous data derived from randomized controlled trials and administrative data, some different findings were retrieved about incidence of VTE. Assessing predictive factors, older age, higher body mass index, and longer disease duration resulted to be significant predictors of VTE occurrence during the follow-up. Taking together these observations, a further evaluation of this issue on specific designed studies is needed to provide more generalizable results to the daily clinical practice.


In vitro selection of a single-stranded DNA molecular recognition element specific for bromacil.
Bromacil is a widely used herbicide that is known to contaminate environmental systems. Due to the hazards it presents and inefficient detection methods, it is necessary to create a rapid and efficient sensing device. Towards this end, we have utilized a stringent in vitro selection method to identify single-stranded DNA molecular recognition elements (MRE) specific for bromacil. We have identified one MRE with high affinity (K d = 9.6 nM) and specificity for bromacil compared to negative targets of selection and other pesticides. The selected ssDNA MRE will be useful as the sensing element in a field-deployable bromacil detection device.


A method for assessing the stability of a membrane protein.
The integral membrane protein diacylglycerol kinase (DGK) from Escherichia coli has been reversibly unfolded in a protein/detergent/mixed micelle system by varying the molar ratio of n-decyl beta-D-maltoside (DM) and sodium dodecyl sulfate (SDS). Unfolding was monitored by circular dichroism (CD) and ultraviolet (UV) absorbance spectroscopy. When unfolding is monitored by measuring changes in absorbance at 294 nm, two distinct denaturation phases are observed, indicative of a stable intermediate. When CD is used as a conformational probe, the resulting denaturation curve contains only one major transition, which corresponds to the first unfolding phase observed by absorbance changes. The unfolding behavior of several mutant proteins in which the tryptophan residues were selectively replaced made it possible to assign the first unfolding phase to a denaturation event in a cytoplasmic domain and the second phase to denaturation of the membrane-embedded portion of the protein. The denaturation curves fit well to a model which assumes two cooperative transitions and a linear relationship between unfolding free energy and SDS concentration. Extrapolation back to zero denaturant indicates an unfolding free energy of 6 kcal/mol for the cytoplasmic domain and 16 kcal/mol for the transmembrane domain. The high apparent stability of the transmembrane domain could explain the high degree of tolerance to amino acid substitutions seen for DGK and other membrane proteins. The approach described in this paper may be applicable to other membrane protein systems.


[Quality of life and attitude to the disease in patients with chronic gastritis].
Examination of 118 patients with various morphofunctional types of chronic gastritis for quality of life (QL) and personal attitude (PA) to the disease has revealed that in non-atrophic gastritis QL deteriorates moderately because of the diet, diagnostic procedures and treatment. PA is anxious, neurastenic, sensitive and hypochondriac; in focal atrophic gastritis QL is the same but PA is less negative, primarily adequate; in diffuse atrophic gastritis there are the worst trends in QL because of the disease, abnormal physical activity and routine life, PA is hypochondric, melancholic, neurastenic and egocentric. Thus, therapeutic policy in various morphofunctional types of chronic gastritis should account for personality reactions of the patients and deterioration of life quality. This allows maintenance of social and medical adaptation of the patients, development of adequate mechanisms of psychological defence, adequate treatment compliance.


Young adult mental health sequelae of eating and body image disturbances in adolescence.
There has been interest in the antecedents and mental health impacts of eating and body image disturbances in adolescence. Less is known about longer-term mental health impacts into young adulthood, as longitudinal studies with data spanning this developmental period are rare. We capitalize on mental health data collected across adolescence and young adulthood from a population-based cohort study that has been following >2000 Australian children and their families from infancy to young adulthood. This sample comprised 1,568 participants who completed the Eating Disorder Inventory drive for thinness and bulimic behavior (the severity of binge-purge patterns) subscales, and a modified version of the body dissatisfaction subscale in mid-adolescence (15-16 years), or the Depression Anxiety Stress Scales in young adulthood (19-20, 23-24, and 27-28 years). After adjusting for baseline demographic and prior mental health factors (<13 years of age), all three indices of eating and body image disturbances in adolescence predicted each mental health outcome in young adulthood. Mental health risks associated with adolescent body dissatisfaction and bulimic behavior scores remained stable across young adulthood, with men having more pronounced problems associated with bulimic behavior scores than women. In contrast, mental health risks associated with adolescent drive for thinness scores diminished across this period similarly for men and women. Findings suggest that adolescent eating and body image disturbances may have long-term mental health impacts that extend into young adulthood. This underscores the need for early preventative intervention, and longer-term monitoring and support for body image and eating disturbances.


Sequential Steps of CRAC Channel Activation.
Interaction between the endoplasmic reticulum protein STIM1 and the plasma membrane channel ORAI1 generates calcium signals that are central for diverse cellular functions. How STIM1 binds and activates ORAI1 remains poorly understood. Using electrophysiological, optical, and biochemical techniques, we examined the effects of mutations in the STIM1-ORAI1 activating region (SOAR) of STIM1. We find that SOAR mutants that are deficient in binding to resting ORAI1 channels are able to bind to and boost activation of partially activated ORAI1 channels. We further show that the STIM1 binding regions on ORAI1 undergo structural rearrangement during channel activation. The results suggest that activation of ORAI1 by SOAR occurs in multiple steps. In the first step, SOAR binds to ORAI1, partially activates the channel, and induces a rearrangement in the SOAR-binding site of ORAI1. That rearrangement of ORAI1 then permits sequential steps of SOAR binding, via distinct molecular interactions, to fully activate the channel.


Considerations on genetic and environmental factors that contribute to resistance or sensitivity of mammals including humans to toxicity of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) and related compounds. Part 1: Genetic factors affecting the toxicity of TCDD.
The marked species differences in short-term toxicity (30-day LD50) of ca. 10,000 (LD50: guinea pigs ca. 1 microgram/kg body wt and Han/Wistar Kuopio rats more than 9600 micrograms/kg body wt) of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) is one of the central issues of the controversies that have developed on the validity of risk assessment strategies for TCDD and related compounds. One of the most challenging issues that toxicologists face today is the identification of genes that contribute to or are responsible for increased resistance or sensitivity to TCDD and related compounds. It is assumed that most, if not all, toxic effects of TCDD are mediated more or less through the binding affinity to the Ah receptor. This hypothesis was extended and tries to explain the differences in sensitivity/resistance of animals including humans to TCDD by their total fat (lipid) content. In this respect the gene or genes which is or are responsible for obesity of mammals including humans are of great interest. An obvious linear positive logarithmic relationship between the oral 30-day LD50 (microgram/kg) of TCDD in different species and strains of mammals and their total body fat content (TBF%) was found: log LD50 = 5.30 x log (TBF)-3.22, or LD50 = 0.000603 x (TBF)5.30. By means of this regression the toxicity of TCDD in mammals including humans of different age and/or body weight can be predicted if their total body fat content is known. Examples of single-gene and polygenic disease models in different mammals, such as nonobese diabetic, diabetic, viable yellow, obese, and fat mice, as well as transgenic mice, and other suitable animal models, such as fatty Zucker rats, Han/Wistar (Kuopio) rats, and minipigs, are discussed, and predicted LD50 values of TCDD in these animals and humans are presented.


Complete amino acid sequence of murine beta 2-microglobulin: structural evidence for strain-related polymorphism.
Primary structural analyses of beta 3-microglobulin isolated from the tumor cell lines EL4.BU (derived from a C57BL/6 mouse) and C14 (derived from a BALB/c mouse) have revealed the presence of an amino acid difference at position 85 of this molecule. beta 2-Microglobulin isolated from histocompatibility antigens of EL4.BU has alanine at this position, whereas that from C14 has aspartic acid. Determination of the sequence of these molecules has employed radiochemical methodology that was developed in studies of murine histocompatibility antigens. The sequence obtained in this study is: Ile - Gln - Lys - Thr - Pro - Gln - Ile - Gln - Val - Tyr - Ser - Arg - His - Pro - Pro - Glu - Asn - Gly - Lys - Pro - Asn - Ile - Leu - Asn - Cys - Tyr - Val - Thr - Gln - Phe - His - Pro - Pro - His - Ile - Glu - Ile - Gln - Met - Leu - Lys - Asn - Gly - Lys - Lys - Ile Pro - Lys - Val - Glu - Met - Ser - Asp - Met - Ser - Phe - Ser - Lys - Asp - Trp - Ser - Phe - Tyr - Ile - Leu - Ala - His - Thr - Glu - Phe - Thr - Pro - Thr - Glu - Thr - Asp - Thr - Tyr - Ala - Cys - Arg - Val - Lys - His - Ala/Asp - Ser - Met - Ala - Glu - Pro - Lys - Thr - Val - Tyr - Trp - Asp - Arg - Asp - Met. Comparison of the sequence of murine beta 2-microglobulin to the sequences reported for the homologues from man, rabbit, and guinea pig indicate identities of 68%, 66%, and 61%, respectively.


Attenuated total reflection Fourier transform infrared spectroscopy for on-chip monitoring of solute concentrations.
We report a cost-efficient Attenuated Total Reflection Fourier Transform Infrared Spectroscopy (ATR-FTIR) method for monitoring concentrations of solutes in solutions flowing through microfluidic channels. The method allows rapid acquisition of spectra and enables chemical characterisation and concentration measurements that are independent of the flow rate of liquids. The method enables independent measurement of concentrations of solutes with distinct spectral features in mixed solutions. For the polymer solutes studied in the present work, the method has a sensitivity of at least 10 microM (0.01 wt%). We also propose the applicability of the method for the differentiation between dissolved and adsorbed amphiphilic species.


Proteomic analysis of porcine oocytes during in vitro maturation reveals essential role for the ubiquitin C-terminal hydrolase-L1.
In this study, we performed proteomic analysis of porcine oocytes during in vitro maturation. Comparison of oocytes at the initial and final stages of meiotic division characterized candidate proteins that were differentially synthesized during in vitro maturation. While the biosynthesis of many of these proteins was significantly decreased, we found four proteins with increased biosynthetic rate, which are supposed to play an essential role in meiosis. Among them, the ubiquitin C-terminal hydrolase-L1 (UCH-L1) was identified by mass spectrometry. To study the regulatory role of UCH-L1 in the process of meiosis in pig model, we used a specific inhibitor of this enzyme, marked C30, belonging to the class of isatin O-acyl oximes. When germinal vesicle (GV) stage cumulus-enclosed oocytes were treated with C30, GV breakdown was inhibited after 28 h of culture, and most of the oocytes were arrested at the first meiosis after 44 h. The block of metaphase I-anaphase transition was not completely reversible. In addition, the inhibition of UCH-L1 resulted in elevated histone H1 kinase activity, corresponding to cyclin-dependent kinase(CDK1)-cyclin B1 complex, and a low level of monoubiquitin. These results supported the hypothesis that UCH-L1 might play a role in metaphase I-anaphase transition by regulating ubiquitin-dependent proteasome mechanisms. In summary, a proteomic approach coupled with protein verification study revealed an essential role of UCH-L1 in the completion of the first meiosis and its transition to anaphase.


Is Mycoplasma synoviae outrunning Mycoplasma gallisepticum? A viewpoint from the Netherlands.
Mycoplasma gallisepticum and M. synoviae are the most relevant mycoplasma species for commercial poultry from the clinical and economic point of view. Although the importance of M. gallisepticum was recognized many decades ago, the relevance of M. synoviae has been a matter of debate. Until the turn of the century, only the respiratory and synovitis forms of the disease were reported, while the majority of infections were subclinical. Since the year 2000 M. synoviae strains with oviduct tropism, able to induce eggshell apex abnormalities and egg drops, have been encountered worldwide. A decreasing incidence of M. gallisepticum has been observed, at least in breeding stock, in countries with control and eradication programmes for this Mycoplasma species. In contrast, the sero-prevalence of M. synoviae is much higher, especially in layer flocks, and in most continents exceeds 70%. Given the emergence of virulent M. synoviae strains with oviduct tropism, its ability to also induce joint and respiratory disease, to act synergistically with other pathogens as well as its much higher sero-prevalence, it seems that M. synoviae is outrunning M. gallisepticum, at least in countries with control and eradication programmes for the latter. This stresses the need to update M. synoviae prevention and control strategies. Thus, in January 2013, the Dutch poultry industry implemented a mandatory control and eradication programme for M. synoviae at all levels of poultry farming with the exception of broilers.


The roles of the ovarian extracellular matrix in fertility.
In the mammalian ovary there is considerable and continuous remodelling of tissue during both fetal and adult life, necessitating changes in extracellular matrix. Matrix is a diverse group of molecules varying in its composition and roles, which include regulation of growth factor activity and cell behaviour. Here we discuss four topical aspects of matrices in ovaries. (1) Our current state of knowledge of latent TGFFbeta binding proteins that can bind the extracellular matrix fibrillins. Fibrillins and latent TGFbeta binding proteins may be very important given the genetic linkage data implicating a role for fibrillin 3 in polycystic ovarian syndrome. They will almost certainly be important in the stromal compartments of the ovary by regulating TGFbeta bioactivity. (2) Follicles which have an unusual ultrastructural follicular basal lamina and poor quality oocytes. The results suggest that the use of oocytes from these follicles should be avoided in assisted reproductive technologies. (3) Evidence that expression of components of focimatrix correlates with expression of aromatase and cholesterol side-chain cleavage in granulosa cells. Focimatrix is a novel type of basal lamina associated with granulosa cells with expression beginning before deviation and continuing until ovulation. It may be involved in maturation of granulosa cells and selection of the dominant follicle. (4) Evidence is presented in support of a hypothesis that follicular fluid accumulates in follicles due to the osmotic potential of hyaluronan and versican, which are matrices produced by granulosa cells and too large to traverse the follicular antrum. These examples illustrate the diversity of matrix and foreshadow potential important discoveries involving extracellular matrix in ovaries.


Term vs. preterm cord blood cells for the prevention of preterm brain injury.
BACKGROUNDWhite matter brain injury in preterm infants can induce neurodevelopmental deficits. Umbilical cord blood (UCB) cells demonstrate neuroprotective properties, but it is unknown whether cells obtained from preterm cord blood (PCB) vs. term cord blood (TCB) have similar efficacy. This study compared the ability of TCB vs. PCB cells to reduce white matter injury in preterm fetal sheep.METHODSHypoxia-ischemia (HI) was induced in fetal sheep (0.7 gestation) by 25 min umbilical cord occlusion. Allogeneic UCB cells from term or preterm sheep, or saline, were administered to the fetus at 12 h after HI. The fetal brain was collected at 10-day post HI for assessment of white matter neuropathology.RESULTSHI (n=7) induced cell death and microglial activation and reduced total oligodendrocytes and CNPase+myelin protein in the periventricular white matter and internal capsule when compared with control (n=10). Administration of TCB or PCB cells normalized white matter density and reduced cell death and microgliosis (P<0.05). PCB prevented upregulation of plasma tumor necrosis factor (TNF)-a, whereas TCB increased anti-inflammatory interleukin (IL)-10 (P<0.05). TCB, but not PCB, reduced circulating oxidative stress.CONCLUSIONSTCB and PCB cells reduced preterm HI-induced white matter injury, primarily via anti-inflammatory actions. The secondary mechanisms of neuroprotection appear different following TCB vs. PCB administration.


Open surgical revision of radial nerve after humeral fracture in polytrauma.
The injury of the radial nerve is not uncommon in humeral shaft fractures. These lesions may be of different types and levels and their treatment changes depending on traumatic characteristic. The authors present a case report of a patient affected by a closed shaft humeral fracture, consequence of an high energy trauma, associated with motor palsy of the radial nerve in which an open surgical revision was performed. The nerve appeared medially displaced and entrapped within the fracture fragments and the fibrous tissue around the lesion and showed signs of posttraumatic suffering without disruption. The nerve was repositioned in its anatomical site after neurolysis and intramedullary nailing of the humeral fracture. The fracture healed after 4 months with full recovery of motor function of the radial nerve.We believe that, in shaft fractures of the humerus with radial nerve deficits, it is imperative to perform surgical exploration of the conditions of the radial nerve itself, in order to obtain a recovery of its functionality.


Educational benefits of the online journal club: A systematic review.
Online journal clubs (JCs) have increased during the COVID-19 pandemic with the resulting social distancing and popularity of online platforms. This systematic review aims to explore current evidence of their use/benefits for clinicians and compare their value to face-to-face (F2F) JCs. PubMed, Web of Science, and Scopus were searched systematically, adhering to PRISMA guidelines. Articles were included if they involved clinicians in medical/surgical populations, using an online JC assessing utility, experience and educational value. Quality assessment was undertaken using MERSQI. Fifteen studies were included with findings synthesised into five themes: critical appraisal skills, satisfaction/value, accessibility/environment, evidence-based practice, and preference of online JC. Studies revealed high satisfaction and equivocal or increased preference of online JCs compared to F2F due to ease of access, diverse participation, and less time/cost spent travelling. Online JCs were found to be educationally valuable, aiding development of critical appraisal skills, and promoting change in practice. Disadvantages included lack of discussion intensity, technical issues, and limited interaction on some platforms. Online JCs are educationally valuable with high satisfaction rates and distinct advantages/disadvantages to F2F JCs. More high-quality studies are required to elucidate the ideal format to further improve their educational value, utility, and adoption.


Understanding the biosynthesis of platelets-derived extracellular vesicles.
Platelet-derived extracellular vesicles (PEVs) are described as sub-cellular vesicles released into circulation upon platelets shear stress, activation, injury, or apoptosis. They are considered as universal biomarkers in a wide range of physiological and pathological processes. They are of tremendous significance for the prediction, diagnosis, and observation of the therapeutic success of many diseases. Understanding their biosynthesis and therefore functional properties would contribute to a better understanding of the pathological mechanisms leading to various diseases in which their levels are raised and they are implicated. The review takes a critical look at the historical background of PEVs, their structural components, the mechanism of their formation, physiological, and exogenous stimuli inducing their release and their detection. It concludes by highlighting on the importance of undertaking in-depth studies into PEVs biosynthesis and subsequently gaining a better understanding of their biological role in general.


Bronchoalveolar lavage cell differential on microscope glass cover. A simple and accurate technique.
We describe a quick and easy technique to perform cell differentials on bronchoalveolar lavage: the microscope glass cover. Lavage fluids of 72 subjects were analyzed by 3 techniques: glass cover, filter, and cytocentrifuge preparations. Seventy-seven other lavages were analyzed by glass cover and cytocentrifuge preparations alone. Data for the 72 subjects studied by all 3 techniques showed that the cell counts on glass cover and filter preparations were similar, e.g., lymphocytes, 19.2% (range, 0.5 to 94%) and 20.9% (range, 3 to 95%), respectively (Spearman's correlation coefficient, 0.98). However, on cytocentrifuge preparations, lymphocyte counts were lower (8.3%; range, zero to 87%) and macrophage counts were higher (p less than 0.005). Comparison of glass cover and cytocentrifuge preparation mixtures with varying amounts (20 to 80%) of purified blood leukocytes labeled with 51Cr (greater than or equal to 72% lymphocytes) showed that a significant amount of radioactive cells was lost during the cytocentrifuge technique in contrast to the glass cover technique. Because neutrophils represented a low proportion of lavage cells, we also evaluated cell suspensions with known neutrophil contents (10 to 70%); we found no difference in neutrophil counts obtained with the 3 techniques. Lavage data analysis of 40 young nonsmoking volunteers showed that glass cover lymphocyte count was also higher than counts on cytocentrifuge preparations: 16.5% (range, 3 to 45%) and 8.2% (range, 2.5 to 35%), respectively. In this group, the distribution of glass cover lymphocyte percentages was normal (p = 0.21, chi 2 test), and the one-tailed 95% confidence interval was 18.6 to 34.7% (mean plus 1.65 standard deviation).(ABSTRACT TRUNCATED AT 250 WORDS)


Human pluripotent stem cells identify molecular targets of trisomy 12 in chronic lymphocytic leukemia patients.
Identifying precise targets of individual cancers remains challenging. Chronic lymphocytic leukemia (CLL) represents the most common adult hematologic malignancy, and trisomy 12 (tri12) represents a quarter of CLL patients. We report that tri12 human pluripotent stem cells (hPSCs) allow for the identification of gene networks and targets specific to tri12, which are controlled by comparative normal PSCs. Identified targets are upregulated in tri12 leukemic cells from a cohort of 159 patients with monoclonal B cell lymphocytosis and CLL. tri12 signaling patterns significantly influence progression-free survival. Actionable targets are identified using high-content drug testing and functionally validated in an additional 44 CLL patient samples. Using xenograft models, interleukin-1 receptor-associated kinase 4 (IRAK4) inhibitor is potent and selective against human tri12 CLL versus healthy patient-derived xenografts. Our study uses hPSCs to uncover targets from genetic aberrations and apply them to cancer. These findings provide immediate translational potential as biomarkers and targets for therapeutic intervention.


Studies on the immunizing capacity of orally administered particulate antigens. II. Production of immunological memory in germ-free mice by orally administered sheep erythrocytes.
A study was perfromed to find out, whether or not the oral administration of sheep erythrocytes results in a general primary immune reaction as well as in effective priming for the secondary response in both conventional and germ-free NMRI mice. Whereas negative results were obtained with conventionally held mice, five oral applications of 0.3 ml of a 60% suspension of sheep erythrocytes to germ-free mice, each dose separated by an interval of 24 hr, resulted in a general primary immune response both at the cellular and humoral levels. When such pretreated mice were given an i.p. injection of 4 times 10(8) sheep erythrocytes as a secondary antigenic stimulus 32 days after the last of the five oral applications, the subsequent response was characterized by the predominant development of 7S hemolysin-producing spleen cells. This evidently indicates that effective priming for the secondary response has taken place by the orally administered antigen.


Frameshift mutations in the bax gene are not involved in development of ovarian endometrioid carcinoma.
The purpose of this study was to determine whether mutations in the Bax gene play a role in the development of ovarian endometrioid carcinoma with a microsatellite instability phenotype. We analyzed a total of 60 tumor specimens, 49 ovarian endometrioid carcinomas and 11 concurrent endometrial endometrioid carcinomas from 49 patients. Fourteen ovarian endometrioid carcinomas and 6 endometrial endometrioid carcinomas showed a microsatellite instability-high phenotype. Tumor and normal-tissue specimens from eight patients with a microsatellite instability-high phenotype colorectal carcinoma were included in this study as controls. The presence or absence of a mutation in the poly (G) 8 tract of the Bax gene was determined by polymerase chain reaction followed by direct DNA sequence analysis. A 1-base pair deletion at the poly (G) 8 tract and no expression of Bax and Bcl-2 proteins were identified in one microsatellite instability-high endometrial endometrioid carcinoma. Immunohistochemical staining for Bax and Bcl-2 proteins was negative on the tumor specimen that had this 1-base pair deletion. No mutations were found in the synchronous microsatellite instability-high ovarian endometrioid carcinoma from the same patient. In contrast, four (50%) of the eight microsatellite instability-high sporadic colorectal carcinomas had a mutation in the poly (G) 8 tract. Although Bax plays an important role in carcinogenesis of the colorectum with microsatellite instability-high phenotype, Bax may not play a direct role in the genesis of ovarian endometrioid carcinoma, regardless of microsatellite instability status.


Cohesin loss alters adult hematopoietic stem cell homeostasis, leading to myeloproliferative neoplasms.
The cohesin complex (consisting of Rad21, Smc1a, Smc3, and Stag2 proteins) is critically important for proper sister chromatid separation during mitosis. Mutations in the cohesin complex were recently identified in a variety of human malignancies including acute myeloid leukemia (AML). To address the potential tumor-suppressive function of cohesin in vivo, we generated a series of shRNA mouse models in which endogenous cohesin can be silenced inducibly. Notably, silencing of cohesin complex members did not have a deleterious effect on cell viability. Furthermore, knockdown of cohesin led to gain of replating capacity of mouse hematopoietic progenitor cells. However, cohesin silencing in vivo rapidly altered stem cells homeostasis and myelopoiesis. Likewise, we found widespread changes in chromatin accessibility and expression of genes involved in myelomonocytic maturation and differentiation. Finally, aged cohesin knockdown mice developed a clinical picture closely resembling myeloproliferative disorders/neoplasms (MPNs), including varying degrees of extramedullary hematopoiesis (myeloid metaplasia) and splenomegaly. Our results represent the first successful demonstration of a tumor suppressor function for the cohesin complex, while also confirming that cohesin mutations occur as an early event in leukemogenesis, facilitating the potential development of a myeloid malignancy.


The Dual Role of USP11 in Cancer.
Ubiquitination is one of the most crucial ways of protein degradation and plays an indispensable role in various living activities of cells. The deubiquitinating enzyme (DUB) is the main practitioner of the reversal of ubiquitination. Up till the present moment, nearly 100 DUBs from six families have been confirmed. USP11 is a member of the largest subfamily of cysteine protease DUBs, involving in the regulation of cell cycle, DNA repair, regulating signaling pathways, tumor development, and other important biological behaviors. This review briefly describes the structure and function of USP11 and comprehensively describes its dual role in tumorigenesis and development, as well as its targeted therapy.


Elucidation of The Anti-Inflammatory Effect of Vanillin In Lps-Activated THP-1 Cells.
Vanillin, a kind of phenolic compound, is naturally found in food and beverage and widely used as a flavoring agent. In view of the safety and universality of vanillin, exploring the functions of vanillin on human is of great value. Thus, lipopolysaccharide (LPS)-activated THP-1 cells were selected as the cell model to evaluate the anti-inflammatory effect of vanillin in this study. On the basis of the results, vanillin markedly suppressed the expression of inflammatory cytokines (that is, TNF-α, IL-1β, IL-6, and IL-8), mediators (NO, iNOS, PGE2, and COX-2), and NLRP3 inflammasome (that is, NLRP3, ASC, and caspase-1), blocked the LPS-induced activation of the NF-κB/IκBα/AP-1 signaling pathway, and activated the gene expression of the Nrf2/HO-1 signaling pathway. In addition, it was confirmed that vanillin was unable to react with LPS due to the results of quantification by HS-SPME-GC-MS. Hence, vanillin could effectively attenuate LPS-induced inflammatory response by regulating the expression of intracellular signaling pathways in THP-1 cells. It is a potent anti-inflammatory component found in food and beverage. These findings might contribute to the overall understanding of the potential health benefits of vanillin for food application. PRACTICAL APPLICATION: In this study, the anti-inflammatory effect of vanillin (VA) was evaluated by ELISA, real-time PCR, and western blot in LPS-induced THP-1 cells. The hypothesis that VA could react with LPS was excluded due to the results of quantification by HS-SPME-GC-MS. On the basis of the result, vanillin could effectively attenuate LPS-induced inflammatory response in THP-1 cells and was a potent anti-inflammatory component natural in food and beverage. These findings might contribute to the overall understanding of the potential health benefits of vanillin for food application.


Treatment of orthostatic hypotension with erythropoietin.
Patients with orthostatic hypotension caused by autonomic neuropathy frequently have a decreased red-cell mass. This would be expected to compromise their effective circulating blood volume and aggravate the orthostatic hypotension. We studied the effect of increasing the red-cell mass with erythropoietin, given subcutaneously in a dose of 50 U per kilogram of body weight three times a week for 6 to 10 weeks to eight patients with orthostatic hypotension--four men, one teenage boy, and three women (age range, 17 to 68 years). Four patients had type I diabetes mellitus and autonomic neuropathy, three patients had pure autonomic failure, and one patient had sympathotonic orthostatic hypotension. Seven patients received fludrocortisone (0.1 or 0.2 mg per day) before, during, and after the trial of erythropoietin. The red-cell volume, plasma volume, and hemodynamic response to orthostatic stress were measured before and after therapy. Erythropoietin increased the mean (+/- SD) hematocrit from 0.34 +/- 0.04 to 0.45 +/- 0.04 (P < 0.005) and increased the red-cell volume from 16.8 +/- 3.9 to 25.3 +/- 3.1 ml per kilogram (P < 0.005), but had no effect on plasma volume. The systolic blood pressure increased from 81 +/- 11 to 100 +/- 24 mm Hg (P < 0.01) and the diastolic blood pressure increased from 46 +/- 10 to 63 +/- 18 mm Hg (P < 0.01) while the patients were standing. The average systolic and diastolic blood pressure while the patients were supine did not increase significantly, although hypertension in the supine position developed in three patients. Orthostatic dizziness improved during treatment in six of the eight patients. In patients with orthostatic hypotension, increasing the red-cell volume with erythropoietin elevates blood pressure while standing. Possible long-term adverse effects are not known.


Dot-probe evaluation of cognitive processing biases in patients with panic disorder: a failure to replicate and extend.
A number of cognitive psychology paradigms have been used to demonstrate that panic disorder patients selectively process stimuli that are related to themes of physical threat and catastrophe. The present investigation was an attempt to replicate and extend initial findings obtained with the dot-probe paradigm, a visual attention paradigm which allows for the assessment of allocation of visual attention to stimuli of varied emotional valence via the measurement of detection latencies for visual probes that follow their presentation. Twenty-two panic disorder patients and 20 healthy control subjects were tested using neutral, social threat, and panic symptom/fear cues. No between-group differences were observed in detection latencies for visual probes that followed neutral, social threat, or panic symptom/fear cues. These results represent a failure to replicate previous reports of selective processing in panic disorder and suggest that panic disorder patients may not selectively process cues related to bodily sensation and panic-related fears. Implications of this finding are discussed.


Interobserver Variation in Cerebellar Tonsillar Tip Localization: Comparison of 3 Different Osseous Landmarks on Magnetic Resonance Imaging.
The aim was to evaluate the interobserver agreement in the assessment of cerebellar tonsil position on sagittal magnetic resonance imaging using 3 different osseous landmarks. This retrospective study consisted of brain magnetic resonance imagings performed at our institution in patients with and without Chiari I malformation between January 2010 and 2012. Sagittal T1-weighted images were reviewed by 2 senior board-certified neuroradiologists (blinded to underlying clinical diagnosis) with measurement of both cerebellar tonsillar positions based on lines drawn perpendicular from the tonsillar tip to the foramen magnum [FM] line, C1 line, and C2 line. Spearman correlation coefficients were calculated. Interobserver variation between the readers was assessed using Bland-Altman analysis and intraclass correlation coefficient. A total of 320 cerebellar tonsils on 160 patients, 50 with Chiari I malformations, and 110 control subjects without Chiari I malformation were evaluated. The Spearman correlation coefficients for the entire cohort were 0.86 (FM), 0.94 (C1), and 0.90 (C2). Bland-Altman analysis for the entire cohort showed the best interobserver agreement for C1 line (-0.3 mm bias) and the least for C2 line (4.6 mm bias). The Intraclass correlation coefficients for all patients were 0.84 (FM), 0.92 (C1), and 0.54 (C2). The least bias and highest correlation coefficients were also seen individually in the Chiari and non-Chiari cohorts with the C1 technique. Determination of cerebellar tonsillar position using a C1 arch landmark may be superior to the currently more commonly used FM-based landmark with lesser interobserver variability and higher interobserver correlation.


Ultrastructural and histochemical study of previtellogenic oogenesis in the desert lizard Scincus mitranus (Squamata, Sauropsida).
The structure of the granulosa in reptilian sauropsids varies between groups. We investigated the follicle development in the desert lizard Scincus mitranus. In the germinal bed, oogonia, and primary oocytes were identified and found to be interspersed between the epithelial cells. Previtellogenesis was divided into three stages: early, transitional, and late previtellogenic stages. During the early previtellogenic stage (diplotene), the oocyte is invested by small epithelia cells that formed a complete single layer, which may be considered as a young follicle. The transitional previtellogenic stage was marked by proliferation and differentiation of the granulosa layer from a homogenous layer consisting of only small cells to a heterogeneous layer containing three cell types: small, intermediate, and large cells. The late previtellogenic stage was marked by high-synthetic activity of large cells and the initiation of cytoplasmic bridges between large granulosa cells and the oocyte. Small cells were the only type of granulosa cells that underwent division. Thus, these cells may be stem cells for the granulosa cell population and may develop into intermediate and subsequently large cells. The intermediate cells may be precursors of large cells, as suggested by their ultrastructure. The ultrastructure of the large granulosa was indicative of their high synthetic activity. Histochemical analysis indicated the presence of cholesterol and phospholipids in the cytoplasm of large cells, the zona pellucida, among the microvilli, in the bridges region, and in the cortical region of the oocyte cytoplasm. These materials may be transferred from large cells into the oocyte through cytoplasmic bridges and provide nutritive function to large cells rather than functioning in steroidogenesis or vitellogenesis.


The role of extrahepatic metabolism in the pharmacokinetics of the targeted covalent inhibitors afatinib, ibrutinib, and neratinib.
Despite the fact that much progress has been made recently in the development of targeted covalent inhibitors (TCIs), their pharmacokinetics (PK) have not been well characterized in the light of extrahepatic clearance (CLextH) by glutathione (GSH)/glutathione S-transferase (GST)-dependent conjugation attributable to the unique electrophilic structure (e.g., acrylamide moiety) of TCI compounds. In the present study, CLextH values were examined in rat, dog, and monkey to predict the contribution of CLextH to the PK of the TCIs afatinib, ibrutinib, and neratinib in humans. Afatinib and neratinib both underwent extensive conjugation with GSH in buffer and cytosol fractions of liver and kidney, whereas ibrutinib showed much lower reactivity/susceptibility to GSH/GST-dependent conjugation. The CLextH in each species was calculated from the difference between observed total body clearance and predicted hepatic clearance (CLH) in cryopreserved hepatocytes suspended in 100% serum of the corresponding species. The power-based simple allometry relating the CLextH for the unbound compound to animal body weight was applicable across species for afatinib and neratinib (R(2) ≥ 0.9) but not for ibrutinib (R(2) = 0.04). The predicted AUC after oral administration of afatinib and neratinib agreed reasonably closely with reported values in phase I dose-escalation studies. Comparisons of CLextH and CLH predicted that CLextH largely determined the PK of afatinib (>90% as a proportion of total body clearance) and neratinib (∼34%) in humans. The present method can serve as one of the tools for the optimization of PK in humans at the discovery stage for the development of TCI candidates.


Implications of the sentinel lymph node procedure for local and systemic adjuvant treatment.
The objective of the sentinel lymph node procedure in breast cancer is to perform an accurate axillary staging and provide good local control, while sparing the patients the morbidity of an axillary lymph node dissection. Since its routine clinical use, questions were raised concerning the implications for local and systemic adjuvant treatment. This review provides an update of the recent literature. As a result of a more detailed histopathologic work-up of the sentinel lymph node, higher rates of lymph node metastases are detected. This leads to an upstaging of a subset of node-negative patients and an increase in the overall percentage of node-positive patients. However, the clinical implications of micrometastases and isolated tumor cells remain unclear. Furthermore, sentinel lymph nodes may be found in the internal mammary lymph node chain but the treatment of these nodes is subject of debate. Current guidelines recommend axillary lymph node dissection in patients with a positive sentinel node. The surgical removal of the internal mammary lymph node is only indicated in the context of a clinical trial. Radiation therapy of the axilla is an acceptable alternative for patients who refuse an axillary lymph node dissection (clinical trial). The value of radiotherapy to the internal mammary lymph node has never been established. Systemic treatment decisions in patients with a macrometastasis or micrometastasis in the sentinel lymph node follow the guidelines of node-positive patients, whereas in patients with isolated tumor cells only, guidelines for node-negative patients are followed. The results of ongoing clinical trials will be important for the development of further guidelines.


A Combination of Guanidyl and Phenyl Groups on a Dendrimer Enables Efficient siRNA and DNA Delivery.
Gene therapy has received considerable attention due to its great potential in the treatment of various diseases; however, the design of efficient and biocompatible carriers for the delivery of siRNA as well as DNA still remains a major challenge. In this study, we developed an efficient carrier for gene delivery by modification of a compound containing both guanidyl and phenyl groups on the surface of a cationic dendrimer. The guanidyl group on the dendrimer facilitates nucleic acid condensation via specific guanidinium-phosphate interactions, whereas the phenyl group on the polymer is critical for efficient endosomal escape. The combination of guanidyl and phenyl shows a synergistic effect in facilitated endocytosis. The designed material is much more efficient in siRNA and DNA delivery than control materials such as dendrimers engineered with a guanidyl or phenyl group only, as well as intact dendrimers, and shows comparable efficacy to commercial transfection reagent Lipofectamine 2000. In addition, the material and its complex with nucleic acid show minimal toxicity on the transfected cells. This study provides a new strategy to develop multifunctional polymers for efficient siRNA and DNA delivery.


Preoperative Serum IL-12p40 Is a Potential Predictor of Kasai Portoenterostomy Outcome in Infants with Biliary Atresia.
The standard-of-care treatment for biliary atresia (BA) is surgical restoration of bile flow by Kasai portoenterostomy. We aimed to study serum interleukin- (IL-) 12p40, a natural antagonist for the proinflammatory IL-12p70, and its relation to surgical outcomes of BA. The study included 75 infants with neonatal cholestasis: BA group (n = 25), non-BA cholestasis group (n = 30), and neglected BA group (n = 20), in addition to thirty healthy neonates serving as controls. IL-12p40 was measured by ELISA in all individuals and a second assessment was performed 3 months postoperatively in the BA group. The surgical outcomes were classified as successful (bilirubin ≤ 2 mg/dl) or failed (bilirubin > 2 mg/dl). IL-12p40 was higher in BA compared to that in the non-BA and control groups (P values were 0.036 and <0.0001, resp.) but comparable to that in the neglected BA group. Preoperative IL-12p40 levels in BA patients were significantly higher in successful Kasai compared with failed Kasai and a cutoff level of 547.47 pg/ml could predict the successful outcome with 87.5% sensitivity and 82.4% specificity. Three-month postoperative IL-12p40 tended to decrease in both the successful and failed groups. In conclusion, preoperative serum IL-12p40 is a potential predictor of Kasai outcome. Serial postoperative measurements may anticipate the failure of an initially successful operation, hence the need for liver transplantation.


Genetics of speciation in the Aedes (Stegomyia) scutellaris subgroup (Diptera:Culicidae). 4. Chromosomal relationships of Aedes cooki with four sibling species.
Morphology and behavior of chromosomes and development of testes and sperm were examined in hybrids from interspecific crosses involving Aedes cooki and four sibling species of the Aedes (Stegomyia) scutellaris subgroup of mosquitoes. The degree of abnormality in hybrid spermatogenesis in interspecific crosses involving Aedes cooki males and females of four sibling species paralleled the geographic distributions of these species and the genetic divergence indicated by other genetic studies. Hybrids from crosses involving Aedes malayensis females and Aedes cooki males were characterized by atrophied testes and extensive chromosome breakage. Hybrids from crosses involving Aedes alcasidi females and Aedes cooki males suggested a possible pericentric inversion distinguishing the largest autosome of Aedes alcasidi from that of Aedes cooki. Hybrids from interspecific crosses involving females of Aedes polynesiensis and Aedes pseudoscutellaris and males of Aedes cooki showed high percentages of univalents of the smallest chromosome pair. Hybrid spermatogenesis in two interspecific crosses involving Aedes cooki females differed from results of reciprocal crosses. Data were scant, however, and interpretation was difficult in view of negligible hatch in all interspecific crosses involving Aedes cooki females.


[Paraparesis of the lower extremities after acute aortic dissection and a thoracic meningioma].
A 70-year-old woman with acute chest pain was admitted to a hospital in stable cardiovascular conditions. The patient had no history of cardio-circulatory disease. An acute myocardial infarction was excluded by ECG and blood tests. A computed tomography (CT) revealed an aortic dissection (Stanford type A) which extended to the left subclavian artery. She was transferred to our institution and underwent urgent operation during which the ascending aorta and the proximal arch were replaced by a prosthesis. A few days after surgery, she developed progressive paresis of both legs. A control CT scan of the aorta revealed no evidence of a persisting aortic dissection. However, magnetic resonance tomography showed a meningioma of the thoracic spinal cord. The patient underwent surgical resection of the meningioma and her neurological symptoms diminished over the next few days. Besides spinal ischemia, paresis of both legs after acute aortic dissection may be caused by rare lesions such as a thoracic meningioma.


Cerebral 18F-FDG PET in macrophagic myofasciitis: An individual SVM-based approach.
Macrophagic myofasciitis (MMF) is an emerging condition with highly specific myopathological alterations. A peculiar spatial pattern of a cerebral glucose hypometabolism involving occipito-temporal cortex and cerebellum have been reported in patients with MMF; however, the full pattern is not systematically present in routine interpretation of scans, and with varying degrees of severity depending on the cognitive profile of patients. Aim was to generate and evaluate a support vector machine (SVM) procedure to classify patients between healthy or MMF 18F-FDG brain profiles. 18F-FDG PET brain images of 119 patients with MMF and 64 healthy subjects were retrospectively analyzed. The whole-population was divided into two groups; a training set (100 MMF, 44 healthy subjects) and a testing set (19 MMF, 20 healthy subjects). Dimensionality reduction was performed using a t-map from statistical parametric mapping (SPM) and a SVM with a linear kernel was trained on the training set. To evaluate the performance of the SVM classifier, values of sensitivity (Se), specificity (Sp), positive predictive value (PPV), negative predictive value (NPV) and accuracy (Acc) were calculated. The SPM12 analysis on the training set exhibited the already reported hypometabolism pattern involving occipito-temporal and fronto-parietal cortices, limbic system and cerebellum. The SVM procedure, based on the t-test mask generated from the training set, correctly classified MMF patients of the testing set with following Se, Sp, PPV, NPV and Acc: 89%, 85%, 85%, 89%, and 87%. We developed an original and individual approach including a SVM to classify patients between healthy or MMF metabolic brain profiles using 18F-FDG-PET. Machine learning algorithms are promising for computer-aided diagnosis but will need further validation in prospective cohorts.


lncRNA Kcnq1ot1 promotes bone formation by inhibiting miR-98-5p/Tbx5 axis in MC3T3-E1 cells.
Long non-coding (lnc)RNA KCNQ1 opposite strand/antisense transcript 1 (Kcnq1ot1) has been shown to regulate multiple biological processes. However, the functional role of Kcnq1ot1 in osteoporosis and the underlying mechanism are still unclear. The present study aimed to investigate the function of lncRNA Kcnq1ot1 in osteogenic differentiation. Alkaline phosphatase (ALP) activity was measured using an ALP assay kit. Western blotting was performed to assess the expression levels of osteogenic differentiation-associated proteins. Reverse transcription-quantitative PCR was performed to detect Kcnq1ot1, microRNA (miR)-98-5p and T-box transcription factor 5 (Tbx5) expression levels. The binding of Kcnq1ot1 with miR-98-5p and that of miR-98-5p with Tbx5 were predicted by starBase and TargetScan databases, respectively, and verified using dual luciferase reporter assays. The mineralization of MC3T3-E1 cells was observed using an Alizarin red S staining assay. The results revealed that expression of Kcnq1ot1 was increased and that of miR-98-5p was decreased during osteogenic differentiation. Additionally, Kcnq1ot1 was shown to target miR-98-5p and inhibit its expression. Inhibiting miR-98-5p reversed the inhibitory effect of Kcnq1ot1 knockdown on osteogenic differentiation and mineralization of MC3T3-E1 cells. Furthermore, Kcnq1ot1 regulated Tbx5 expression via miR-98-5p. Overexpressing miR-98-5p or downregulating Tbx5 expression reversed the promotive effect of Kcnq1ot1 overexpression on osteogenic differentiation and mineralization of MC3T3-E1 cells. In conclusion, these findings suggested that Kcnq1ot1 may promote bone formation by inhibiting miR-98-5p and upregulating Tbx5. Kcnq1ot1, miR-98-5p and Tbx5 may therefore serve as promising targets for the treatment of osteoporosis.


Second-line chemotherapy and beyond for non-small-cell lung cancer.
Platinum-based chemotherapy is now established in the treatment of all stages of lung cancer. Despite the benefits of therapy, the majority of patients treated for lung cancer will ultimately progress. Some will retain good performance status and be candidates for additional, tumor-directed treatment. Docetaxel and pemetrexed have documented activity in phase III trials in patients progressing after first-line, platinum-based therapy. Gefitinib has recently been approved for treatment of disease in the third line. Numerous other agents with diverse mechanisms are currently undergoing evaluation in this setting. This review evaluates the population eligible for such treatments and discusses recent trials in second and subsequent lines of therapy.


Successful cardiopulmonary bypass in diabetics with anaphylactoid reactions to protamine.
Two insulin dependent diabetics with previous anaphylactic like (anaphylactoid) reactions to protamine underwent successful cardiopulmonary bypass for coronary artery surgery. Platelet concentrates instead of protamine were used to neutralise their systemic heparinisation. In both cases the anaphylactoid reactions first became apparent after administration of protamine sulphate at the end of cardiac catheterisation. These cases show that adverse reactions to protamine need not be a contraindication to cardiopulmonary bypass and cardiac surgery and emphasise that this condition should be considered in all patients with a history of previous protamine exposure or one which may be associated with anaphylactoid reactions to protamine.


Comprehensive analysis of an immune-related ceRNA network in identifying a novel lncRNA signature as a prognostic biomarker for hepatocellular carcinoma.
The function of competitive endogenous RNA (ceRNA) network in the immune regulation of hepatocellular carcinoma (HCC) is unclear. Our study aimed to construct an immune-related ceRNA network and develop an immune-related long noncoding RNA (lncRNA) signature to assess the prognosis of HCC patients and to optimize the treatment methods. We firstly constructed a ceRNA regulatory network for HCC using differentially expressed lncRNAs, mRNAs and microRNAs (miRNAs) from the Cancer Genome Atlas. A signature was constructed by 11 immune-related prognostic lncRNAs from the ceRNA network. The survival analysis and receiver operating characteristic analysis validated the reliability of the signature. Multivariate Cox regression analysis revealed that the signature could act an independent prognostic indicator. This signature also showed high association with immune cell infiltration and immune check blockades. LINC00491 was identified as the hub lncRNA in the signature. In vitro and in vivo evidence demonstrated that silencing of LINC00491 significantly inhibited HCC growth. Finally, 59 lncRNAs, 21 miRNAs, and 26 mRNAs were obtained to build the immune-related ceRNA network for HCC. In conclusion, our novel immune-related lncRNA prognostic signature and the immune-related ceRNA network might provide in-depth insights into tumor-immune interaction of HCC and promote better individual treatment strategies in HCC patients.


Diallyl trisulfide exerts anti-inflammatory effects in lipopolysaccharide-stimulated RAW 264.7 macrophages by suppressing the Toll-like receptor 4/nuclear factor-κB pathway.
Diallyl trisulfide (DATS; di‑2‑propen‑1‑yl trisulfide) is an organic polysulfide compound found in garlic and other allium vegetables. Although certain studies have demonstrated that DATS possesses strong anti‑inflammatory activity, the underlying molecular mechanisms remain largely unresolved. In the present study, the anti‑inflammatory potential of DATS was investigated using the murine macrophage RAW 264.7 cell model. At non‑toxic concentrations, DATS inhibited the production of nitric oxide (NO) and prostaglandin E2 by inhibiting inducible NO synthase and cyclooxygenase‑2 expression at the transcriptional level in lipopolysaccharide (LPS)‑activated RAW 264.7 macrophages. DATS attenuated the release of the pro‑inflammatory cytokines, tumor necrosis factor‑α and interleukin‑1β, by inhibiting mRNA expression, respectively. DATS also suppressed LPS‑induced DNA‑binding activity of nuclear factor‑κB (NF‑κB), as well as the nuclear translocation of the NF‑κB p65, which correlated with the inhibitory effects of DATS on inhibitor κB (IκB) degradation. In addition, DATS was observed to significantly suppress LPS‑induced Toll‑like receptor 4 (TLR4) and myeloid differentiation factor 88 expression and the binding of LPS to macrophages, indicating the antagonistic effect of DATS against TLR4. Furthermore, blocking TLR4 signaling with the specific TLR4 signaling inhibitor, CLI‑095, increased the anti‑inflammatory potential of DATS in LPS‑stimulated RAW 264.7 macrophages. These data demonstrate that DATS may attenuate the initiation of LPS‑mediated intracellular signaling cascades by suppressing activation of NF‑κB and by inhibiting binding of LPS to TLR4 on macrophages.


Evaluation of immunoperoxidase-stained tissue sections with an electrophoresis densitometer.
The application of the Phoroscope electrophoresis densitometer for the objective evaluation of immunoperoxidase-stained tissue sections at the light microscope level is described. The instrument directs a beam of light through the specimen (Kodachrome photomicrograph slides) and projects an absorbance pattern (as peaks and valleys) on an oscilloscope screen. The height of a peak is proportional to the optical density of a stained area, while the width of the peak is proportional to the diameter of an area. The utility of the densitometer was tested in sections of rat pituitary gland immunostained with NIAMDD antiserum against rat prolactin to reveal prolactin cells and sections of rat ventral prostate pretreated with NIAMDD rat prolactin and then immunostained to reveal Golgi region-localized prolactin binding sites in epithelial cells. The densitometer quantitated (by relative absorbance) the changes in immunostaining in serial sections of rat pituitary gland reacted with different dilutions of prolactin antiserum and in serial sections of rat ventral prostate pretreated with different concentrations of rat prolactin. In addition, the densitometric pattern provided information regarding the frequency and pattern of immunostaining within a particular microscopic field. Potential uses of an electrophoresis densitometer for the quantitation and standardization of immunohistochemistry are discussed.


Defining the noninferiority margin and analysing noninferiority: An overview.
Noninferiority trials are used to assess whether the effect of a new drug is not worse than an active comparator by more than a noninferiority margin. If the difference between the new drug and the active comparator does not exceed this prespecified margin, noninferiority can be concluded. This margin must be specified based on clinical and statistical reasoning; however, it is considered as one of the most challenging steps in the design of noninferiority trials. Regulators recommend that the margin should be defined based on the historical evidence of the active comparator (the latter is often the well-established standard treatment of the disease), which can be performed by different approaches. There are several factors and assumptions that need to be accounted for during the process of defining the margin and during the analysis of noninferiority. Three methods are commonly used to analyse noninferiority trials: the fixed-margin method; the point-estimate method; and the synthesis method. This article provides an overview of analysing noninferiority and choosing the noninferiority margin.


ε-Poly-l-Lysine/plasmid DNA nanoplexes for efficient gene delivery in vivo.
The present work addresses the development and characterization of ε-Poly-l-Lysine/pDNA polyplexes and evaluation for their improved transfection efficacy and safety as compared to polyplexes prepared using Poly-l-Lysine and SuperFect®. Self-assembling polyplexes were prepared by varying the N/P ratio to obtain the optimum size, a net positive zeta potential and gel retardation. The stability in presence of DNase I and serum was assured using gel retardation assay. Their appreciable uptake in MCF-7 and 3.5, 3.79 and 4.79-fold higher transfection compared to PLL/pDNA polyplexes and 1.60, 1.53 and 1.79-fold higher transfection compared to SuperFect®/pDNA polyplexes in MCF-7, HeLa and HEK-293 cell lines respectively, affirmed the enhanced transfection of ε-PLL/pDNA polyplexes which was well supported with in vivo transfection and gene expression studies. The <8% in vitro hemolysis and >98% viability of MCF-7, HeLa and HEK-293 cells in presence of ε-PLL/pDNA polyplexes addressed their safety, which was also ensured using in vivo toxicity studies, where hemocompatibility, unaltered levels of biochemical markers and histology of vital organs confirmed ε-PLL to be an effective and safer alternative for non-viral genetic vectors.


The role of 5-alpha-reductase inhibition as monotherapy in view of the MTOPS data.
Medical treatment for the symptoms of benign prostatic hyperplasia (BPH) consists of a blockers and 5-alpha-reductase inhibitors. Data suggest that 5-alpha-reductase inhibitors can prevent progression of BPH and reduce the risk of BPH-related surgery, especially in men with large-volume prostates. Results from the largest randomized, prospective, placebo-controlled trial, the Medical Therapy of Prostatic Symptoms trial, have been presented. These results support the notion of using 5-alpha-reductase inhibitors for the prevention of BPH progression and BPH-related surgery. Furthermore, long-term 5-alpha-reductase inhibitor monotherapy, although slow in onset, is a viable therapy for symptom relief in men with mild to moderate symptoms.


Sclerocorneal graft and sequential removal of melted cornea after severe corneal burn with perforation.
Corneal burn grade IV usually leads to blindness. Several different surgical techniques remain challenging owing to the extensive tissue damage. Here, we introduce a novel technique with a 15 mm corneoscleral and limbal homologous graft combined with sequential autologous corneal removal ab interno, with a vitrectomy probe to save the anterior chamber angle. In vivo anatomy with optical coherence tomography is the surgical key. A large 15 mm sclerocorneal graft is sutured on top of the remainder of the destroyed cornea and sclera after removal of the epithelium and conjunctiva, with anterior synechiolysis if necessary, peripheral iridectomy and conjunctivoplasty. The recipient central corneal stroma is not removed, primarily to protect the anterior chamber angle. After three weeks, the collagenolytic central recipient corneal stroma can be removed with a small 23 g vitrectomy probe, respecting the lens and scleral spur. The corneoscleral graft remains clear under systemic and local immunosuppression. Intraocular pressure is well controlled because the anterior chamber angle is respected. Recurrent corneal erosions need close follow-up. Therapeutic soft contact lenses can support topical therapy. In cases of sclercorneal graft decompensation or rejection after 3-5 years, a new sclerocorneal graft (with limbal donation) seems to be superior to perforating keratoplasty without limbal stem cell transplantation. Repeated sclerocorneal grafts after severe corneal burn with limbal transplantation and maintenance of the complete anterior angle structure are a successful option for preventing blindness and achieving good visual acuity. Clin. Anat. 31:39-42, 2018. © 2017 Wiley Periodicals, Inc.


Effect of nintedanib on airway inflammation and remodeling in a murine chronic asthma model.
Nintedanib is a multi-tyrosine kinase receptor inhibitor recently approved for treatment of idiopathic pulmonary fibrosis. Although angiogenesis is a key process involved in airway structural changes in patients with bronchial asthma, the effect of nintedanib targeting the angiokinase pathway on airway inflammation and remodeling has not been evaluated. We used a 3-month ovalbumin (OVA) challenge mouse model of airway remodeling. Nintedanib was orally administrated during the challenge period, and the effects were examined based on the percentage of airway inflammatory cells, airway hyper-reactivity (AHR), peribronchial goblet cell hyperplasia, total lung collagen and smooth muscle area. The expression of growth factor receptors was analyzed in mice lung tissues. The OVA challenged group showed a significant increase in airway eosinophilic inflammation, elevated Th2 cytokines, AHR, and airway remodeling compared to those in the control group. The airway remodeling process, as evaluated by goblet cell hyperplasia, total lung collagen level, and airway smooth muscle area, was suppressed by nintedanib compared to that by OVA. Nintedanib effectively suppressed the phosphorylation of vascular endothelial growth factor/ platelet derived growth factor subunit2/fibroblast growth factor3 receptors in the mice lung. Nintedanib effectively ameliorated airway inflammation and remodeling in an OVA-induced chronic asthma model. These results suggest that nintedanib could be a new treatment agent targeting airway remodeling in patients with severe asthma.


Transient Potassium Channels: Therapeutic Targets for Brain Disorders.
Transient potassium current channels (IA channels), which are expressed in most brain areas, have a central role in modulating feedforward and feedback inhibition along the dendroaxonic axis. Loss of the modulatory channels is tightly associated with a number of brain diseases such as Alzheimer's disease, epilepsy, fragile X syndrome (FXS), Parkinson's disease, chronic pain, tinnitus, and ataxia. However, the functional significance of IA channels in these diseases has so far been underestimated. In this review, we discuss the distribution and function of IA channels. Particularly, we posit that downregulation of IA channels results in neuronal (mostly dendritic) hyperexcitability accompanied by the imbalanced excitation and inhibition ratio in the brain's networks, eventually causing the brain diseases. Finally, we propose a potential therapeutic target: the enhanced action of IA channels to counteract Ca2+-permeable channels including NMDA receptors could be harnessed to restore dendritic excitability, leading to a balanced neuronal state.