Estrogen and progesterone receptors in human breast cancer. Correlation with histologic subtype and degree of differentiation.
Microscopic review of 490 consecutive human breast biopsy and mastectomy specimens were correlated with estrogen and progesterone receptor content of the tissue, by subtype and degree of differentiation. Of the 4 grades of differentiation, the less differentiated Grade III and IV tumors showed significantly lower levels of estrogen and progesterone receptors in infiltrating ductal and lobular carcinoma (P less than 0.001). In contrast, patients with medullary carcinoma had the lowest tissue levels of estrogen and progesterone receptors with approximately 80% of the cases with less than 10 fmol/mg protein. Patients with mucinous carcinoma had the highest percentages of positive estrogen and progesterone receptor levels (75% and 87%, respectively). Sixty-three percent of the patients with Grade IV infiltrating ductal carcinoma were younger than 53 years of age (P less than 0.001). Patients younger than 53 years of age with Grade II and III infiltrating ductal carcinoma also had significantly lower levels of estrogen receptors, but not of progesterone receptors, than those patients older than 53 years of age (P less than 0.001). Nineteen of 20 "normal" breast tissue specimens were negative (less than 3 fmol/mg protein) for estrogen and progesterone receptors. About 50% of 17 tissue specimens from benign breast lesions (fibroadenoma, fibrocystic disease, sclerosing adenosis) showed positive estrogen (greater than 10 fmol/mg protein) or progesterone receptor values. In two patients with gynecomastia, no estrogen or progesterone receptors were detectable.


The effects of glycine-glutamine dipeptide replaced l-glutamine on bovine parthenogenetic and IVF embryo development.
Relative to alanine and serine amino acid levels, glutamine is highly abundant in follicular fluid, and is an important source of energy required for oocyte maturation and embryo development. Thus, glutamine is an essential component of in vitro embryo culture media. However, glutamine has poor stability and degrades spontaneously in solution to form ammonia and pyrrolidonecarboxylic acid. In the present study, we aimed to explore the effect of substituting l-glutamine with glycine-glutamine, a more stable glutamine, on development of early parthenogenetic embryos and in vitro fertilization (IVF) embryos in bovine. Results revealed that glycine-glutamine can significantly increase cleavage rate (parthenogenetic embryos:87.24% vs. 72.61%, IVF embryos:89.33% vs. 83.79%, P < 0.01), blastocyst number (parthenogenetic embryos:24.98% vs. 18.07%, IVF embryos:33.53% vs. 27.29%, P < 0.01), and blastocyst number (parthenogenetic embryos:96 vs. 76, IVF embryos:114 vs. 109, P < 0.01), reduce blastocyst apoptosis (parthenogenetic embryos:3.72% vs. 6.65%, IVF embryos:2.53% vs.6.23%, P < 0.01), alleviate embryo ammonia toxicity, and reduce the content of reactive oxygen species (ROS) compared with the l-glutamine. In addition, glycine-glutamine can alter epigenetic reprogramming by increasing the expression of HDAC1 (Histone Deacetylase 1) and decreasing the relative expression levels of H3K9 acetylation in early parthenogenetic embryos and IVF embryos. From our present study, we concluded that glycine-glutamine is an effective substitute of glutamine in modified synthetic oviduct fluid with amino acids (mSOFaa).


Experimental model of autoimmune hemolytic anemia induced in mice with levodopa by intraperitoneal injection or oral feeding.
In this paper we describe a murine experimental model of autoimmune hemolytic anemia induced with multiple injections or oral feeding of levodopa. Strain A mice were intraperitoneally injected or fed with levodopa, at a dose equivalent to the one used in human therapy, and subsequently they developed cycles of IgM, IgG and IgA anti-mouse red blood cells (MRBC) autoantibody responses. Levodopa injection induced serum IgM and IgG anti-MRBC responses and levodopa feeding enhanced the serum anti-MRBC IgA response. The appearance of autoantibodies in the serum was followed by binding of the autoantibodies to mice erythrocytes and three phases of anemia. Red cell bound IgM and IgG autoantibodies were predominant in levodopa-injected mice whereas red cell bound IgA autoantibodies were predominant in levodopa-fed mice. The specificity of the serum IgA autoantibody was not restricted since it interacted with erythrocytes of various species.


Generation of estradiol within the pregnant guinea pig uterine compartment with special reference to the myometrium.
Between about 50 and 58 days of gestation, the guinea pig chorion becomes attached in its entirety to the uterine wall, suggesting a facilitation of transfer of agents such as steroids between these tissues. At a time between 59 and 64 days, relaxation of the pubic symphysis starts, and anywhere from 5 to 8 days after that event delivery takes place. The present in vitro study was undertaken to evaluate estrone sulfate as a substrate for local production of estradiol, via the action of estrogen sulfatase and 17-hydroxysteroid dehydrogenase, in chorion, endometrium and myometrium taken at four distinct stages of gestation, as follows: 50 days, representing pre-chorion attachment to the uterus (stage 50); 1 or 2 days before pubic symphysis relaxation (minus 1 day, or -1 day); 1 day following relaxation (+1 day); and 1-2 days before delivery (late, or L). At these same stages, the metabolite patterns formed from estradiol were evaluated for endometrium and myometrium. Each of the tissues behaved somewhat differently. Overall hydrolysis of estrone sulfate by endometrium and myometrium exceeded that by chorion. Generation of free steroid from estrone sulfate increased 3-fold in chorion between stages 50 and -1 and during this period estradiol production from estrone sulfate increased 9-fold and continued to rise until delivery. Cytosolic estrogen sulfotransferase activity of chorion decreased 7-fold between stages 50 and -1. This suggested a tissue environment geared to producing potentially active estradiol. However, myometrium converted very little estrone into estradiol until just before delivery despite the facile formation of estrone from estradiol at stages -1, +1 and L. The control of estrogen metabolism by interaction of tissues in the uterus and by some form of enzyme regulation in these tissues suggests a possible role for locally produced estrogen in the stages leading up to parturition.


Catalytic and non-catalytic amidation of carboxylic acid substrates.
The present review offers an apt summary of amide bond formation with carboxylic acid substrates by taking advantage of several methods. Carboxamides can be regarded as a substantial part of organic and medicinal chemistry due to their utility in synthesizing peptides, lactams, and more than 25% of familiar drugs. Moreover, they play a leading role in the synthesis of bioactive products with anticancer, antifungal, and antibacterial properties. The data are arranged based on the type and amount of reagents used to conduct amidation and are also divided into the following categories: catalytic amidation of carboxylic acids, non-catalytic amidation, and transamidation.


Critical role of ADP interaction with P2Y12 receptor in the maintenance of alpha(IIb)beta3 activation: association with Rap1B activation.
Platelet integrin alpha(IIb)beta3 plays a crucial role in platelet aggregation, and the affinity of alpha(IIb)beta3 for fibrinogen is dynamically regulated. Employing modified ligand-binding assays, we analyzed the mechanism by which alpha(IIb)beta3 maintains its high-affinity state. Washed platelets adjusted to 50 x 10(3) microL(-1) were stimulated with 0.2 U mL(-1) thrombin or 5 microm U46619 under static conditions. After the completion of alpha(IIb)beta3 activation and granule secretion, different kinds of antagonists were added to the activated platelets. The activated alpha(IIb)beta3 was then detected by fluorescein isothiocyanate (FITC)-labeled PAC1. The addition of 1 mum AR-C69931MX (a P2Y12 antagonist) or 1 mm A3P5P (a P2Y1 antagonist) disrupted the sustained alpha(IIb)beta3 activation by approximately 92% and approximately 38%, respectively, without inhibiting CD62P or CD63 expression. Dilution of the platelet preparation to 500 microL(-1) also disrupted the sustained alpha(IIb)beta3 activation, and the disruption by such dilution was abrogated by the addition of exogenous adenosine 5'-diphosphate (ADP) in a dose-dependent fashion. The amounts of ADP released from activated platelets determined by high-performance liquid chromatography were compatible with the amounts of exogenous ADP required for the restoration. We next examined the effects of antagonists on protein kinase C (PKC) and Rap1B activation induced by 0.2 U mL(-1) thrombin. Thrombin induced long-lasting PKC and Rap1B activation. AR-C69931MX markedly inhibited Rap1B activation without inhibiting PKC activation. Our data indicate that the continuous interaction between released ADP and P2Y12 is critical for the maintenance of alpha(IIb)beta3 activation.


Overexpression of LSD1 in hepatocellular carcinoma: a latent target for the diagnosis and therapy of hepatoma.
The aim of this study was to investigate the expression of LSD1 in human hepatocellular carcinoma (HCC) cell lines and HCC samples. In this study, we examined LSD1 expression in 60 paired liver cancer tissues and adjacent noncancerous tissues by Western blot. In addition, we analyzed LSD1 expression in 198 HCC samples by immunohistochemistry. The relationship between LSD1 expression and clinicopathological features was investigated. The HCC cell line SMMC-7721 was transfected with LSD1 siRNA expressing plasmids. We subsequently examined in vitro cell growth using the MTT assay and anchorage-independent growth through a soft-agar colony-formation assay. In addition, the expression levels of Bcl-2 and c-Myc were also examined. Immunohistochemistry and Western blotting consistently confirmed LSD1 overexpression in HCC tissues compared with adjacent non-neoplastic tissues (P < 0.01). Additionally, immunostaining showed more LSD1-positive cells in the higher tumor stage (T3-4) and tumor grade (G3) than in the lower tumor stage (T1-2, P < 0.001) and tumor grade (G1-2, P < 0.001). Knockdown of LSD1 expression in HCC cells led to decreased cell proliferation. The expression of Bcl-2 and c-Myc were down-regulated after transfection of LSD1 siRNA into HCC cell line SMMC-7721. In conclusion, because LSD1 was overexpressed in HCC and has an important role in the development of HCC, LSD1 could be a latent target in the diagnosis and therapy of HCC.


Modulation of H+/H- exchange in iridium-hydride 2-hydroxypyridine complexes by remote Lewis acids.
A series of iridium hydride complexes featuring dihydrogen bonding are presented and shown to undergo rapid H+/H- exchange (1240 s-1 at 25 °C). We demonstrate that the H+/H- exchange rate can be modified by post-synthetic modification at a remote site using BH3, Zn(C6F5)2, and [Me3O][BF4]. This route provides a complementary strategy to traditional methods that rely on pre-metalation modifications to a metal's primary sphere.


Exploring health literacy in patients with chronic kidney disease: a qualitative study.
Patients with chronic kidney disease make day-to-day decisions about how to self-manage their disease. Chronic kidney disease (CKD) includes a risk for progression towards end-stage renal disease and the development of comorbidities, such as cardiovascular disease, which represents the leading cause of death among these patients. To reduce these risks, CKD patients are recommended to follow a healthy lifestyle with physical activity, food and fluid restrictions, and adherence to complex medication regimes throughout all phases of the disease. To manage the complexity of this health situation, health literacy (HL) is considered essential. The current prevailing understanding is that HL is a multidimensional concept and comprises a range of cognitive, affective, social, and personal skills that determine the motivation and ability to gain access to, understand, and use health information. Recently, we investigated multiple aspects of HL in CKD patients in a quantitative cross-sectional study utilizing the Health Literacy Questionnaire (HLQ) and observed that finding good health information and appraising health information were the most challenging aspects of HL. This study aimed to explore CKD patients' lived experiences of different dimensions of HL presented in the HLQ. This qualitative study utilized in-depth semistructured interviews. Twelve patients with different levels of HL were included. The interviews were analyzed using thematic analysis as described by Braun and Clarke. We identified three main themes that were significant for CKD patients' HL: 1. Variation in people's attitudes and behavior as health information seekers, 2. The problem of fragmented healthcare in the context of multimorbidity makes the healthcare system challenging to navigate, and 3. The value of a good relationship with healthcare providers. CKD patients take different approaches to health information. Limiting or avoiding health information may be a strategy used by some individuals to cope with the disease and does not necessarily mean that health information is inaccessible or difficult to understand. Comorbidity and a fragmented healthcare system can make the healthcare system challenging to navigate. A good and trusting relationship with healthcare providers seems to promote several aspects of HL and should be promoted to optimize CKD patients' HL.


N-Doped Carbon Fibers Derived from Porous Wood Fibers Encapsulated in a Zeolitic Imidazolate Framework as an Electrode Material for Supercapacitors.
Developing highly porous and conductive carbon electrodes is crucial for high-performance electrochemical double-layer capacitors. We provide a method for preparing supercapacitor electrode materials using zeolitic imidazolate framework-8 (ZIF-8)-coated wood fibers. The material has high nitrogen (N)-doping content and a specific surface area of 593.52 m2 g-1. When used as a supercapacitor electrode, the composite exhibits a high specific capacitance of 270.74 F g-1, with an excellent capacitance retention rate of 98.4% after 10,000 cycles. The symmetrical supercapacitors (SSCs) with two carbon fiber electrodes (CWFZ2) showed a high power density of 2272.73 W kg-1 (at an energy density of 2.46 W h kg-1) and an energy density of 4.15 Wh kg-1 (at a power density of 113.64 W kg-1). Moreover, the SSCs maintained 81.21% of the initial capacitance after 10,000 cycles at a current density of 10 A g-1, which proves that the SSCs have good cycle stability. The excellent capacitance performance is primarily attributed to the high conductivity and N source provided by the zeolite imidazole framework. Because of this carbon material's unique structural features and N-doping, our obtained CWFZ2 electrode material could be a candidate for high-performance supercapacitor electrode materials.


Liraglutide effects on glycemic control and weight in patients with type 2 diabetes Mellitus: A real-world, observational study and brief narrative review.
Glycemic control and weight gain are two essential considerations in the pharmacological management of type 2 diabetes mellitus. Pharmacological agents are effective in lowering blood glucose levels but may result in significant weight gain. Liraglutideeffectively maintains glycemic control while reducingweight. This is a real-world study and brief narrative review of the effects of liraglutide on glycemic control and weight in adult patients with type 2 diabetes mellitus. The study uses data extracted from the electronic health record of the Ministry of National Guard-Health Affairs. In this study of 348 subjects, there was a statistically significant reduction in hemoglobin A1c of 0.9% (P < .0001) and weight of 2.3 kg (P < .0001). The majority (77.3%) were on concomitant insulin.Subjects with a baseline hemoglobin A1c greater than 9% had a significantly greater reduction than those below 9% (-0.7%;P < .0001). Those with a weight more than 100 kg had a significantly greater reduction than those below 100 kg (-0.9 kg;P = .0096). In this real-world, observational study, liraglutide was shown to be effective in improving glycemic control and reducing weight in adult patients with type 2 diabetes mellitus.


Laparoscopic hysterectomy with manual morcellation of the uterus: an original technique that permits the safe and quick removal of a large uterus.
We present an original technique to rapidly extract a large uterus during laparoscopic hysterectomy. Manually morcellating the uterus is a safe and effective technique that overcomes the technical difficulties associated with traditional electrical morcellators.


A merocyanine-based dual-mode optical probe for detection of hydrazine and its bioimaging application in vitro and vivo.
In this paper, a Merocyanine-based turn-on probe (McyA) has been developed for colorimetric and fluorescent dual-mode detection of N2H4 via an intra-molecular charge transfer (ICT) mechanism. In the presence of N2H4, the probe shows an obvious chromogenic response and fluorescent enhancement. Based on this feature, the synthesized McyA can be applied to quantify N2H4 concentration from 0.12 to 5.0 μM and 0.5-10 μM with a detection limit of 0.042 μM (1.3 ppb, S/N = 3) and 0.140 μM (4.5 ppb, S/N = 3), respectively. Moreover, McyA has also be successfully employed for imaging analysis of N2H4 distribution in different organs from the N2H4 ingested mice model, revealing the potential application of McyA as a powerful fluorescent sensor for tracking detection and risk assessment of hydrazine in vivo.


Highly Boosting Circularly Polarized Luminescence of Chiral Metal-Imidazolate Frameworks.
To develop a simple and general method for improving the circularly polarized luminescence (CPL) performances of materials is of great significance. In this work, two pairs of CPL-active homochiral metal-organic frameworks (MOFs) P/M-Et and P/M-Et(Cd) with eta topology are reported. In comparison to the reported isomorphic Zn-imidazolate MOFs P-Me and M-Me, both luminescence dissymmetry factor (glum ) and photoluminescence quantum yields (ΦPL ) of P-Et and M-Et are largely improved by simply changing the methyl group to an ethyl group of ligands in P-Et and M-Et. Furthermore, the |glum | values are significantly amplified up to 0.015 from 0.0057 by introducing the non-luminescent halogenated aromatics, while an enhanced fluorescence efficiency is observed simultaneously (from 27.2% to 47.3%). The figure of merit value is about 40 times larger than that of P-Me and M-Me. Similarly, the CPL performances of P/M-Et(Cd) are improved by about five times after encapsulating fluorobenzene molecules. This work represents a new and simple method for developing CPL-active MOF materials.


Consequences and mechanisms of action of fish-borne toxicants: what we do not know and why.
We summarize and discuss issues that emerged from a workshop entitled 'Human and Animal Neurotoxicology-From Exposure to Great Lakes Pollutants' that was held as part of the May 2000 Annual Meeting of the International Association of Great Lakes Research in Cornwall, Ontario, Canada. The most apparent and distressing finding was the lack of communication that currently exists between scientists of different sub-disciplines-including in vitro and in vivo toxicologists, epidemiologists and risk assessors. Although all participants agreed that integrated, multi-disciplinary research is the only avenue that offers a realistic chance of identifying, characterizing and eventually preventing the adverse consequences of developmental exposure to neurological and reproductive toxicants, there was little consensus on how to achieve this goal. We leave to the readers of this Special Issue of Environmental Toxicology and Pharmacology the daunting task of implementing the greater degree of communication needed to begin to solve problems due to developmental exposure to environmental contaminants.


Investigation of substituent effect on O-C bond dissociation enthalpy of methoxy group in meta- and para-substituted anisoles.
This paper is focused on the theoretical investigation of O-C Bond Dissociation Enthalpy (BDE) of methoxy OCH3 group in 15 meta- and 15 para-substituted anisoles in gas phase, non-polar environment, and water. Density Functional Theory (DFT) calculations were carried out using M06-2X functional and 6-311++G(d,p) basis set. Obtained BDEs were correlated with Brown and Okamoto σp+ and Hammett σm constants representing commonly used descriptors of electron-donating or electron-withdrawing substituent effect. Obtained linear dependences allow the prediction of substituent effect on BDE using σp+ and σm constants. Calculated reaction enthalpies were also compared with available experimental and theoretical ab initio G4 values. Found results suggest that employed method may provide reliable thermochemistry data for demethylation of naturally occurring (poly)phenolic compounds, as well. In all studied environments, substituent induced changes in O-C BDE can be considered equal to those observed for the dissociation of phenolic O-H bond of substituted phenols.


The perceived reality and environment of the mid-career nurse scientist.
A qualitative study was designed to explore facilitators and barriers to success from the perspectives of self-identified mid-career nurse scientists. Data were collected from 84 respondents using open-ended items as part of an electronic survey of self-identified mid-career nurse scientists who were members of regional nursing research societies. Two main themes emerged: "Perceived Realities" and "What's Needed to Achieve Career Success." Mid-career nurse scientists experience both joy from teaching and research roles and dissatisfaction with research support; difficulty balancing heavy teaching workload, administrative responsibilities, and research, and ineffective mentoring. Based on the challenges in their environment, respondents identified numerous ways to enhance career success including (1) increasing or enhancing release time for research activities, (2) formal infrastructure to support research activities, and (3) targeted, effective mentoring by senior scientists.


Biosorption of nickel(II) ions onto Sargassum wightii: application of two-parameter and three-parameter isotherm models.
Biosorption of nickel(II) ions from aqueous solution onto Sargassum wightii has been studied and the equilibrium isotherms were determined. The experimental data obtained at different pH conditions (pH 3.0-4.5) have been analyzed using five two-parameter models (Langmuir, Freundlich, Temkin, Dubinin-Radushkevich and Flory-Huggins) and five three-parameter models (Redlich-Peterson, Sips, Khan, Radke-Prausnitz and Toth). In order to determine the best fit isotherm, three error analysis methods were used to evaluate the data: correlation coefficient, residual root mean square error and chi-square test. The error analysis demonstrated that the three-parameter models better described the nickel biosorption data compared to two-parameter models. In particular, Toth equation provided the best model for nickel biosorption data at all pH conditions examined.


Overview of the HMO movement.
This article provides a short historical review of the HMO movement to set the stage for current HMO developments. It notes the underlying causes of the proliferation of HMO managed care models. It defines what should be the rudiments of a quality HMO program and how the changing societal environment will bring new pressures to bear on HMO programs of the future. As related to the future, the author outlines a direction for national health care reforms. The article concludes that the health care emphasis of the 90s should be in returning some of the responsibility of health care to individual Americans through self-care programs that focus on behavioral change, in turn, leading to healthier Americans, the keys to containing out of control health care costs.


Activation of human lymphocytes in vitro by membrane-damaging toxins from Staphylococcus aureus.
Purified staphylococcal alpha-, gamma-, and delta-toxin were shown to cause activation of lymphoid cells from adult human donors and of cord cells in vitro as measured by [14C]thymidine incorporation after 7 days of incubation. T cell-enriched and T cell-depleted lymphocyte suspensions were activated in a similar fashion. Beta-toxin, on the other hand, exerted no valid stimulation of the various lymphocyte preparations. The lymphocyte-activating properties of alpha- and gamma-toxin were shown to be independent of their hemolytic capacity. The results probably reflect unspecific mitogen effects, but a component of specific reactivity cannot be excluded. We suggest that the unspecific triggering of lymphocytes in vitro is caused by surface-active properties of the toxins.


Modifying Methionine on Proteins.
Methionine has been recognized as an ideal target for labeling proteins without disturbing their tasks. However, exploration in single post-transcriptional modification of methionine is sluggish. In this Highlight, we summarize some of the most exciting reports on the precise control of protein function by selectively modifying methionine residues.


A web of interactions at the ends.
The synthesis of telomeric DNA by telomerase entails repeated cycles of reverse transcription on a short RNA template. In this issue of Molecular Cell, Robart and Collins (2011) describe a set of interactions between human telomerase RNA, protein domains, and the substrate DNA that drives the intricate reaction cycle.


Risk of hemolysis in Plasmodium vivax malaria patients receiving standard primaquine treatment in a population with high prevalence of G6PD deficiency.
Primaquine is essential for the radical cure of Plasmodium vivax malaria, but it poses a potential danger of severe hemolysis in G6PD-deficient (G6PDd) patients. This study aimed to determine whether primaquine is safe in a population with high G6PD prevalence but lacking G6PD diagnosis capacity. In Myanmar, 152 vivax patients were gender- and age-matched at 1:3 for G6PDd versus G6PD-normal (G6PDn). Their risk of acute hemolysis was followed for 28 days after treatment with the standard chloroquine and 14-day primaquine (0.25 mg/kg/day) regimen. Patients anemic and non-anemic at enrollment showed a rising and declining trend in the mean hemoglobin level, respectively. In males, the G6PDd group showed substantially larger magnitudes of hemoglobin reduction and lower hemoglobin nadir levels than the G6PDn group, but this trend was not evident in females. Almost 1/3 of the patients experienced clinically concerning declines in hemoglobin, with five requiring blood transfusion. The standard 14-day primaquine regimen carries a significant risk of acute hemolytic anemia (AHA) in vivax patients without G6PD testing in a population with a high prevalence of G6PD deficiency and anemia. G6PD testing would avoid most of the clinically significant Hb reductions and AHA in male patients.


Central nervous system involvement in patients with rotavirus gastroenteritis.
Two children with rotavirus gastroenteritis are presented. The first case developed a fatal Reye's syndrome and the other one encephalitis with slow recovery. The rotavirus diagnosis was made in both cases by electron microscopy and a significant rise in antibody titres to Nebraska calf diarrhea virus was seen in one of the two patients.


Economic burden of impairment in children with severe or difficult-to-treat asthma.
The cost associated with asthma impairment in children with severe asthma has not been determined. To assess the asthma cost burden in children with severe or difficult-to-treat asthma based on asthma impairment. Children aged 6 to 12 years in The Epidemiology and Natural History of Asthma: Outcomes and Treatment Regimens study with available data at baseline (n = 628), month 12 (n = 385), and month 24 (n = 280) corresponding to the National Heart, Lung, and Blood Institute asthma guidelines' impairment domain were included. Children were categorized as either very poorly controlled (VPC), not well controlled (NWC), or well controlled (WC) and assessed cross-sectionally and longitudinally. Mean total asthma costs based on direct (medication usage, unscheduled office visits, emergency department visits, hospitalizations) and indirect (school/work days lost) asthma costs were assessed. Mean annual total asthma costs were more than twice as high in the VPC group compared with NWC and WC groups (baseline: $7,846, $3,526, $3,766.44, respectively; month 12: $7,326, $2,959, $2,043, respectively; month 24: $8,879, $3,308, $1,861, respectively (all P < .001). Indirect costs accounted for approximately half the total asthma costs for VPC asthma patients at each time point. Significantly lower costs were observed for patients whose impairment status improved or temporarily improved from VPC after baseline. The economic burden of severe or difficult-to-treat asthma in children is associated with VPC asthma and improvement in asthma control and is associated with reducing cost. Further attention to patients with poorly controlled asthma, through better management strategies or more effective medications, may significantly reduce this burden of illness.


Multi-facets of neutrophil extracellular trap in infectious diseases: Moving beyond immunity.
Neutrophil extracellular traps (NETs) are networks of extracellular chromosomal DNA fibers, histones, and cytoplasmic granule proteins. The release of NET components from neutrophils is involved in the suppression of pathogen diffusion. Development of NETs around target microbes leads to disruption of the cell membrane, eventuating in kind of cell death that is called as NETosis. The very first step in the process of NETosis is activation of Nicotinamide adenine dinucleotide phosphate (NADPH) oxidase upon signaling by innate immune receptors. Afterwards, produced Reactive oxygen species (ROS) trigger protein-arginine deiminase type 4, neutrophil elastase, and myeloperoxidase to generate decondensed chromatin and disrupted integrity of nuclear membrane. Subsequently, decondensed chromatin is mixed with several enzymes in the cytoplasm released from granules, leading to release of DNA and histones, and finally formation of NET. Several reports have indicated that NETosis might contribute to the immune responses through limiting the dissemination of microbial organisms. In this review, we discuss recent advances on the role of neutrophils, NETs, and their implications in the pathogenesis of microbial infections. Additionally, the prospective of the NET modulation as a therapeutic strategy to treat infectious diseases are clarified.


Effect of imagery ability on letter-level and word-level processing.
In a letter-identification task, subjects matched a probe letter to the initial letter of a subsequently presented probe word. We varied word frequency and predicted that the performance of vivid imagers would resemble that of typical young adults, whereas the performance of poor imagers would fall in between that of typical young adults and of older adults, or would simply resemble the performance of typical older adults (a linear decrease in reaction time [RT] with word frequency; see Allen & Madden, 1989). The in-between function for young and older adults combined predicts a dip in RT for very-high-frequency words compared to medium-high-, low-, and very-low-frequency words. As predicted, vivid imagers exhibited increased latencies for medium-high-frequency words relative to the other three word-frequency categories, whereas poor imagers exhibited a dip.


Primary Anterolateral Ligament Rupture in Patients Requiring Revision Anterior Cruciate Ligament Reconstruction: A Retrospective Case-Control Magnetic Resonance Imaging Review.
To compare the initial rate of anterolateral ligament (ALL) injury at the time of anterior cruciate ligament (ACL) rupture in patients who subsequently experienced ACL reconstruction graft failure versus patients who did not experience subsequent ACL reconstruction graft failure. Our institution's electronic medical record database was queried for patients who underwent primary ACL reconstruction with subsequent ACL graft rupture. Exclusion criteria included unavailable MRI scan, chronic ACL injury, multi-ligamentous injury, previous ACL reconstruction, and age younger than 13 or older than 50 years. Each patient was paired with an age-, gender-, and graft-matched control who underwent ACL reconstruction without subsequent graft rupture. Each patient was diagnosed with an intact, partially injured, or fully ruptured ALL on initial MRI. The location of ALL injury was also noted. The incidence and location of ALL rupture were compared using χ2 analysis. 1,967 patients underwent primary ACL reconstruction. 128 patients experienced ACL graft rupture, and 55 patients (43%) had MRI scans available for review. 39 of these patients fulfilled inclusion criteria and were matched with a control patient. In the revision group, the ALL was diagnosed as intact, partially torn, and completely torn in 17, 14, and 8 patients, respectively, compared to 18, 13, and 8 patients, respectively in the control group. No difference was found in frequency of ALL rupture (Pearson χ2 = 0.066; P = .968) or rupture location (Pearson χ2 = 4.00, P = 0.135). The incidence of initial ALL injury as documented on MRI was not different in patients who experienced subsequent ACL graft rupture compared with patients who did not experience ACL graft rupture after primary ACL reconstruction. The ALL was more commonly injured on the tibial side in patients with ACL graft rupture and femoral-sided lesions were more common in control patients. Level III, prognostic case-control study.


Protocol for High-Resolution Mapping of Splicing Products and Isoforms by RT-PCR Using Fluorescently Labeled Primers.
We describe an RT-PCR protocol that allows high-resolution mapping of splicing products and isoforms using fluorescently labeled primers. Each species contains one fluorescent group allowing a direct comparison of the different isoforms despite size differences. A custom-size ladder enables the precise determination of cDNA lengths and discrimination of isoforms differing by less than five nucleotides on polyacrylamide gels. This protocol also allows the detection of products from in vitro splicing reactions, circumventing the need to use radiolabeled transcripts. For complete details on the use and execution of this protocol, please refer to Gabunilas and Chanfreau (2016).


Cyclooxygenase product inhibition with acetylsalicylic acid slows disease progression in the Han:SPRD-Cy rat model of polycystic kidney disease.
Renal cyclooxygenase (COX) derived eicosanoids are elevated and lipoxygenase (LOX) products are reduced in the Han:SPRD-Cy rat model of polycystic kidney disease (PKD). Selective COX2 inhibition reduces kidney disease progression, but COX1 levels also are elevated in this model. Since the effect of reducing the products of both COX isoforms and the role of LOX products is not known, weanling normal and diseased Han:SPRD-cy littermates were given either low dose acetylsalicylic acid (ASA), nordihydroguaiaretic (NDGA) or no treatment for eight weeks. Renal eicosanoids were altered in the diseased compared to normal cortex, with COX products being higher and LOX products being lower. ASA reduced COX products, cyst growth and kidney water content, while NDGA reduced LOX products without altering disease progression or kidney function. Hence, a human equivalent ASA dose equal to less than one regular strength aspirin per day slowed disease progression, while further reduction of LOX products did not worsen disease progression.


Sequential modifications of glycans by linkage-specific alkylamidation of sialic acids and permethylation.
Permethylation is useful for glycosidic linkage analysis, but is often accompanied by a large proportion of by-products, especially for glycans containing sialic acids (Sia). Unlike hydroxyl groups of glycans, which are converted to stable methyl ethers by permethylation, the carboxylic acids on Sia are converted to methyl esters, which are easily reversible to carboxylate under alkaline conditions. To overcome this problem, we used linkage-specific alkylamidation to protect Sia prior to the permethylation. This method not only decreased the levels of by-products, but also enabled us to distinguish isomers of α2,3- and α2,6-Sia while simultaneously determining other glycosidic linkages.


Bacteremia and urogenital infection with Actinomyces urogenitalis following prolonged urinary retention.
A case of bacteremia with the fastidious bacteria Actinomyces urogenitalis following lengthy urinary retention is reported in a sixty-year-old man. In 2013, the first case of bacteremia due to A. urogenitalis was presented, secondary to a tubo-ovarian abscess following transvaginal oocyte retrieval. To the best of our knowledge, this is the first male bacteremic episode involving A. urogenitalis related to a urinary tract focus. The patient had no prior urogenital medical history. Extensive susceptibility testing was done on isolates from urinary and blood cultures. The organism exhibited fluoroquinolone resistance but was susceptible to most other antibiotics used in the treatment of urinary infections. Due to its unusual growth requirements infections with A. urogenitalis are most likely an underdiagnosed entity.


Real-time biofeedback to target risk of anterior cruciate ligament injury: a technical report for injury prevention and rehabilitation.
Biofeedback training enables an athlete to alter biomechanical and physiological function by receiving biomechanical and physiological data concurrent with or immediately after a task. To compare the effects of 2 different modes of real-time biofeedback focused on reducing risk factors related to anterior cruciate ligament injury. Randomized crossover study design. Biomechanics laboratory and sports medicine center. Female high school soccer players (age 14.8 ± 1.0 y, height 162.6 ± 6.8 cm, mass 55.9 ± 7.0 kg; n = 4). A battery of kinetic- or kinematic-based real-time biofeedback during repetitive double-leg squats. Baseline and posttraining drop vertical jumps were collected to determine if either feedback method improved high injury risk landing mechanics. Maximum knee abduction moment and angle during the landing was significantly decreased after kinetic-focused biofeedback (P = .04). The reduced knee abduction moment during the drop vertical jumps after kinematic-focused biofeedback was not different (P = .2). Maximum knee abduction angle was significantly decreased after kinetic biofeedback (P < .01) but only showed a trend toward reduction after kinematic biofeedback (P = .08). The innovative biofeedback employed in the current study reduced knee abduction load and posture from baseline to posttraining during a drop vertical jump.


In-column "click" preparation of hydrophobic organic monolithic stationary phases for protein separation.
Two types of macroporous organic polymer monoliths based on glycidyl methacrylate (GMA), 4-vinylbenzyl chloride (VBC) and divinylbenzene (DVB) were prepared inside stainless-steel tubes. Azide functionalities were firstly introduced on the surfaces of poly(GMA-co-DVB) and poly(VBC-co-DVB) monoliths to provide reactive sites for click chemistry. With the application of copper(I)-catalyzed (3 + 2) azide-alkyne cycloaddition, an in-column click-modification approach for covalent attachment of long alkyl chains onto polymer monoliths was developed. The column morphology and surface chemistry of the fabricated monolithic columns were characterized by the scanning electron microscopy, mercury intrusion porosimeter, Fourier transform infrared spectroscopy, and elemental analyses, respectively. The chromatographic performances of the "clicked" stationary phases were demonstrated with the high separation efficiency for a variety of proteins within 4 min.


Combining olfactory test and motion analysis sensors in Parkinson's disease preclinical diagnosis: a pilot study.
Preclinical diagnosis of Parkinson's disease (PD) is nowadays a topic of interest as the neuropathological process could begin years before the appearance of motor symptoms. Several symptoms, among them hyposmia, could precede motor features in PD. In the preclinical phase of PD, a subclinical reduction in motor skills is highly likely. In this pilot study, we investigate a step-by-step method to achieve preclinical PD diagnosis. We used the IOIT (Italian Olfactory Identification Test) to screen a population of healthy subjects. We identified 20 subjects with idiopathic hyposmia. Hyposmic subjects underwent an evaluation of motor skills, at baseline and after 1 year, using motion analysis sensors previously created by us. One subject showed significant worsening in motor measurements. In this subject, we further conducted a dopaminergic challenge test monitored with the same sensors and, finally, he underwent [123 I]-FP/CIT (DaTscan) SPECT brain imaging. The results show that he is probably affected by preclinical PD. Our pilot study suggests that the combined use of an olfactory test and motor sensors for motion analysis could be useful for a screening of healthy subjects to identify those at a high risk of developing PD.


Tonic fibres in axial muscle of cyprinid fish larvae: their definition, possible origins and functional importance.
Teleost fish are known to develop small populations of muscle fibres that are assumed to be tonic in nature although their contractile properties and many other characteristics remain unknown. Here we attempt to resolve some of the ambiguity and confusions surrounding the definition and functional role of tonic fibres in teleosts and provide new information on their ontogeny. We investigate the differentiation of tonic muscle fibres in three species of cyprinid fish using electron microscopy, histochemistry, immunohistochemistry and in situ hybridization. The fine structure of the fibres defined as tonic in the larvae used in this study complies with patterns known from studies in teleost adults. This allows formal definition of tonic fibres in cyprinid larvae. The tonic fibres may be recognized by a variety of features: (1) by their characteristic position along the medial confines of the red muscle insertion at the horizontal septum, (2) their fine structure, including solid clusters of irregularly cleaved myofibrils, thick and wavy Z-lines, and T-tubules at the A-band/I-band transitions, (3) their histochemical features, specifically weak but obvious staining for mATPase after alkaline preincubation, and lack of SDH activity in the more advanced larval stages, (4) their unique immunological properties, being the only fibre type in the myotome that reacts with a serum against chicken tonic myosin (anti- T2). Expression of tonic characters usually begins within a few fibres in the dorsal domain of the superficial red muscle insertion at the horizontal septum and hence involves a high degree of dorso-ventral polarity. The present evidence indicates that tonic fibres arise from separate myogenic stem cells rather than by transdifferentiation from existing red fibres. First appearance of tonic fibres during ontogeny correlates closely with the onset of free swimming and exogeneous feeding. We use this fact to argue that tonic fibres are probably a prerequisite for efficient locomotory control during prey capture, shoaling and predator avoidance.


Chapter 3 Current Challenges in Immature Oocyte Cryopreservation.
Current freezing technology, especially the vitrification method, has markedly improved oocyte survival rate after warming, and the pregnancy rate is comparable to that achieved with fresh oocytes. However, most groups report using oocytes matured in vivo for vitrification. Although immature oocytes can be vitrified successfully, clinical outcomes do not reach that of vitrification of matured oocytes. The current literature suggests that oocytes should be vitrified at mature metaphase II (M-II) stage following IVM rather than at the immature germinal vesicle (GV) stage, because the potential for oocyte maturation is reduced when vitrification is performed on immature oocytes at the GV stage.


Mechanical properties of denture base resin cross-linked with methacrylated dendrimer.
The aim of this study was to investigate the mechanical properties of denture base resin cross-linked with methacrylated dendrimer. The test specimens (3 mm × 10 mm × 65 mm) were fabricated from autopolymerizing resin with the powder/liquid ratio of 10 g/7 ml. The monomer liquid of resin was applied with the mixture of methylmethacrylate and crosslinker dendrimer (DD1) or crosslinker ethyleneglycol dimethacrylate (EGDMA) with five different volume percentages (vol%). The dendrimer crosslinker in this study is a methacrylated molecule (MW=3617 g/mol) with 12 methacrylate groups. Quantity of crosslinkers varied from 1.1 to 9.1 vol%. The specimens (n=8/group) were polymerized in distilled water maintained at 55 °C under pressure of 0.4 MPa for 20 min. Test specimens were stored dry at room temperature before testing. The flexural strength (MPa) and flexural modulus (GPa) was measured with three-point bending test at a crosshead speed of 5mm/min. Surface microhardness (MHN) of matrix area of polymer (n=8/group) was measured with a load of 245.3 mN by 10s. Data were analyzed with two-way ANOVA. ANOVA showed that the addition of DD1 had a significantly higher effect (p<0.05) on flexural modulus and hardness of matrix area than EGDMA but on flexural strength (p>0.05). The effect of quantity differences of crosslinker was statistically significant only on flexural strength (p<0.05). The results of this study suggest that dendrimer-crosslinked resin gives better stiffness than that of EGDMA.


Expression of alpha-dystroglycan correlates with tumour grade and predicts survival in oral squamous cell carcinoma.
Dystroglycan (DG) is a non-integrin adhesion molecule connecting the extracellular matrix to the actin cytoskeleton. Decreased expression of DG has been reported in several human cancers and related to tumour aggressiveness. Expression of the alpha-DG subunit was evaluated by immunostaining in a series of oral squamous cell carcinoma (OSCC) and its relation with traditional prognostic indicators and with the clinical outcome of the patients was evaluated. Alpha-DG expression was easily detected in normal epithelium with a mean percentage of positive cells >80% but was undetectable in a significant fraction (59%) of OSCC. Loss of alpha-DG staining correlated with higher tumour grade (p = 0.04) and stage (p = 0.01), with nodal involvement (p = 0.001) and with an increased risk of recurrence (p = 0.002) and death (p = 0.004) in a univariate analysis, but it was not confirmed as an independent predictor of clinical outcome in a multivariate analysis. Loss of alpha-DG expression, which corresponds to loss of a functional DG complex, is a frequent event in human OSCC. Further studies are warranted on the role of this molecule in the entire multistep process of oral squamous tumorigenesis.


Combining spirometry and fractional exhaled nitric oxide improves diagnostic accuracy for childhood asthma.
There are insufficient evidences supporting the use of spirometric indices along with tests for airway inflammation to improve diagnostic accuracy for asthma. We aimed to study the utility of combination of spirometric indices and fractional exhaled nitric oxide (FeNO) measured at the initial visit in diagnosing asthma. Consecutive children aged 8-16 years who were referred for evaluation of possible asthma were included. At referral, all participants completed FeNO measurements and spirometry. The diagnosis of asthma was established with conventional criteria. Diagnostic performance of the spirometric indices and FeNO was determined using receiver-operator characteristic (ROC) curve analyses. Of 275 participants, 191 children were diagnosed with asthma and showed lower spirometric indices and higher FeNO than non-asthmatics. In the ROC curve analyses, forced expiratory flow between 25% and 75% of vital capacity (FEF25-75 ) percent predicted demonstrated diagnostic performance with the area under the ROC curve (AUC) value of 0.81 (95% CI: 0.76-0.87) which was significantly higher than those for forced expiratory volume in the first second (FEV1 ) percent predicted and FEV1 /forced vital capacity. The combined use of FEF25-75 percent predicted and FeNO improved the AUC to 0.90 (95% CI: 0.86-0.93). In addition, compared to FEF25-75 percent predicted or FeNO alone, this combination improved sensitivity with comparable specificity. FEF25-75 percent predicted had a better diagnostic value in detection of childhood asthma than other standard spirometric indices and its combination with FeNO improves the diagnostic accuracy for childhood asthma.


The extended chondrocyte lineage: implications for skeletal homeostasis and disorders.
Endochondral bone formation relies on a finely controlled sequence of chondrocyte proliferation, maturation and hypertrophy that establishes the growth plate which, combined with the deposition of bone upon the cartilage template, mediates longitudinal skeletal growth. Recent lineage studies support a chondrocyte-osteoblast differentiation continuum and the presence of skeletal stem cells within cartilage. The biological significance of the lineage extension and the mechanisms controlling the process are unclear. In this review, we describe recent work on the extended chondrocyte-osteoblast lineage and its contribution to the development, growth and repair of bone and to bone disorders that provides insight into the process and the molecular controls involved. The implications for skeletal homeostasis are discussed.


Color biogenesis data of tomatoes treated with hot-water and high temperature ethylene treatments.
Controlled postharvest stresses were used to induce the synthesis of carotenoids in tomato fruit. The accumulation of carotenoids was observed by the change of color of the tomato fruit from green to red. This change of color was monitored by the a* value and hue of the CIELAB* color coordinates in which the a* value increased following a sigmoidal curve and hue decreased in a similar trend. This sigmoidal curve marked the transition from chloroplasts to chromoplasts; in other words, the change of color tracked the disorganization or degreening, which was simultaneously accompanied by chromoplast biogenesis or red color development when tomatoes were at the Turning stage of development. The color data and photographic images provides information on how heat stress affected the synchronicity of chloroplast disorganization and chromoplast biogenesis in the early developmental stages of tomato ripening.


Formation of recombinant DNA of bacteriophage lambda by recA function of Escherichia coli without duplication, transcription, translation, and maturation.
Genetic recombination of phage lambda DNA mediated by Rec function of Escherichia coli was studied in the absence of duplication, transcription, translation, and maturation. Cells were jointly infected with double amber mutants, lambda D-F-I and lambda S-R-, and incubated in the presence of chloramphenicol and rifampin. The am+ recombinant DNA molecules formed within the cell were detected by in vitro packaging as viable recombinant phages. This system was used to measure the recombination activity of rec- bacteria. In recA or recA recB bacteria, the number of recombinant DNA molecules was about 1% of the rec+ level. In contrast, almost normal numbers of recombinant DNA molecules were formed in recB or recC cells. Therefore, (1) the recombination mediated by recA function does not need de novo protein synthesis; all gene products required for the recombination are present in the cell. (2) It can occur without duplication, transcription, and maturation of recombining DNA molecules. (3) The ATP dependent DNase (exonuclease V) controlled by recB and recC genes is not required for formation of recombinant DNA molecules.


Perlecan Domain V induces VEGf secretion in brain endothelial cells through integrin α5β1 and ERK-dependent signaling pathways.
Perlecan Domain V (DV) promotes brain angiogenesis by inducing VEGF release from brain endothelial cells (BECs) following stroke. In this study, we define the specific mechanism of DV interaction with the α(5)β(1) integrin, identify the downstream signal transduction pathway, and further investigate the functional significance of resultant VEGF release. Interestingly, we found that the LG3 portion of DV, which has been suggested to possess most of DV's angio-modulatory activity outside of the brain, binds poorly to α(5)β(1) and induces less BEC proliferation compared to full length DV. Additionally, we implicate DV's DGR sequence as an important element for the interaction of DV with α(5)β(1). Furthermore, we investigated the importance of AKT and ERK signaling in DV-induced VEGF expression and secretion. We show that DV increases the phosphorylation of ERK, which leads to subsequent activation and stabilization of eIF4E and HIF-1α. Inhibition of ERK activity by U0126 suppressed DV-induced expression and secretion of VEGR in BECs. While DV was capable of phosphorylating AKT we show that AKT phosphorylation does not play a role in DV's induction of VEGF expression or secretion using two separate inhibitors, LY294002 and Akt IV. Lastly, we demonstrate that VEGF activity is critical for DV increases in BEC proliferation, as well as angiogenesis in a BEC-neuronal co-culture system. Collectively, our findings expand our understanding of DV's mechanism of action on BECs, and further support its potential as a novel stroke therapy.


How do heterogeneous R&D investments affect China's green productivity: Revisiting the Porter hypothesis.
Currently, China's environmental pollution control investment and R&D investment have increased at a remarkable speed, exceeding the GDP growth rate. Against this background, determining the impact of different orientations of R&D investment on green total-factor productivity (GTFP) is essential for overcoming the dual dilemma of resource depletion and environmental degradation. By applying the perpetual inventory method (PIM), meta-frontier DEA method and mediation effect test methods, this paper empirically tests the effects and mechanisms of environmentally induced R&D (ER&D) and traditional R&D (TR&D) on promoting GTFP during 2004-2019 in China at the provincial level. The results show that (1) ER&D investment has significantly promoted the growth of GTFP, while TR&D's promoting effects on GTFP are not significant; (2) ER&D promotes the growth of GTFP through the three channels of emission reduction, clean energy consumption, and green technology progress; and (3) from the perspective of regional heterogeneity, ER&D's promoting effects on GTFP in the eastern provinces are higher than the central and western provinces, and the promoting effects in the northern provinces are higher than in the southern provinces. From the perspective of pollution degree heterogeneity, ER&D's promotion of GTFP shows an inverted U-shaped characteristic as the pollution level increases. This means that, in the process of China's environmental governance, it is necessary to increase ER&D investment and guide green innovation to serve pollution control to achieve sustainable and high-quality economic development.


A new interpeduncular screw fixation system: biomechanics, radiologic appearances and complications of the Steffee spine-plate implant.
A newly developed spinal fixation system, designed to assist spinal fusion, is described and case histories are reported to illustrate some radiologic appearances and complications. The biomechanical concepts underlying its use are briefly presented, with the advantages of transpeduncular fixation being emphasized.


A comparative study of the inhaled dry powders of salbutamol and fenoterol and their delivery systems.
A study was designed to establish whether there was a difference in response to inhaled salbutamol powder (400 micrograms) and fenoterol powder (200 micrograms) and whether the specific inhaler devices were a contributing factor. In the study of 40 children no significant difference could be attributed to the devices, but the salbutamol treated group had a marginally better response in two parameters of lung function.


Engineering lower inhibitor affinities in β-D-xylosidase of Selenomonas ruminantium by site-directed mutagenesis of Trp145.
β-D-Xylosidase/α-L-arabinofuranosidase from Selenomonas ruminantium is the most active enzyme reported for catalyzing hydrolysis of 1,4-β-D-xylooligosaccharides to D-xylose. One property that could use improvement is its relatively high affinities for D-glucose and D-xylose (K (i) ~ 10 mM), which would impede its performance as a catalyst in the saccharification of lignocellulosic biomass for the production of biofuels and other value-added products. Previously, we discovered that the W145G variant expresses K(i)(D-glucose) and K(i)(D-xylose) twofold and threefold those of the wild-type enzyme. However, in comparison to the wild type, the variant expresses 11% lower k(cat)(D-xylobiose) and much lower stabilities to temperature and pH. Here, we performed saturation mutagenesis of W145 and discovered that the variants express K (i) values that are 1.5-2.7-fold (D-glucose) and 1.9-4.6-fold (D-xylose) those of wild-type enzyme. W145F, W145L, and W145Y express good stability and, respectively, 11, 6, and 1% higher k(cat)(D-xylobiose) than that of the wild type. At 0.1 M D-xylobiose and 0.1 M D-xylose, kinetic parameters indicate that W145F, W145L, and W145Y catalytic activities are respectively 46, 71, and 48% greater than that of the wild-type enzyme.


The seizure prediction characteristic: a general framework to assess and compare seizure prediction methods.
The unpredictability of seizures is a central problem for all patients suffering from uncontrolled epilepsy. Recently, numerous methods have been suggested that claim to predict from the EEG the onset of epileptic seizures. In parallel, new therapeutic devices are in development that could control upcoming seizures provided that their onset is known in advance. A reliable clinical application controlling seizures, consisting of a seizure prediction method and an intervention system, would improve patient quality of life. The question therefore arises as to whether the performance of the seizure prediction methods is already sufficient for clinical applications. The answer requires assessment criteria to judge and compare these methods, but recognized criteria still do not exist. Based on clinical, behavioral, and statistical considerations, we suggest the "seizure prediction characteristic" to evaluate seizure prediction methods. Results of this approach are exemplified by its application to the "dynamical similarity index" seizure prediction method using 582 hours of intracranial EEG data, including 88 seizures.


Using existing data to estimate aquifer properties, Great Lakes Region, USA.
To determine specific storage and porosity, areally limited and time-consuming aquifer tests are frequently done. Hydrogeologic studies often do not have the resources to collect such data and rely on existing data sources for aquifer properties. An alternative tool for determining these aquifer properties is the analysis of earth tides. The objective of this study was to determine whether existing water-level and barometric-pressure data could be used to determine aquifer properties, such as porosity and specific storage, on a regional scale. In this study, national databases from the Great Lakes Region were queried for continuous records of groundwater-level and barometric-pressure data. Records from 37 selected wells were then analyzed for barometric efficiency and earth-tide responses. Specific-storage (S(s) ) and porosity values were determined, and the quality of the results were assessed with a measure of the "goodness of fit" (percent variance) of reconstruction of the response. Records from wells completed in several aquifer systems were analyzed with varying degrees of success. Aquifer S(s) values ranging from 5.9 × 10(-8) to 3.8 × 10(-6) /m were derived, with percent variance of reconstruction ranging from 1% to 78%. Comparisons with aquifer and laboratory testing of S(s) and porosity are favorable if the percent variance of reconstruction is above about 30%. Although the earth-tide-analysis method is not suitable for every situation, the S(s) and porosity of aquifers can, in many places, be estimated with existing water-level and barometric-pressure data or with data that are relatively inexpensive to collect.


Phenolic derivatives from the rhizomes of Dioscorea nipponica and their anti-neuroinflammatory and neuroprotective activities.
Dioscorea nipponica (Dioscoreaceae) have been used as traditional medicines for diabetes, inflammatory and neurodegenerative diseases in Korea. The aim of the study was to isolate the bioactive components from the rhizomes of Dioscorea nipponica and to evaluate their anti-neuroinfalmmatory and neuroprotective activities. The phytochemical investigation of 50% EtOH extract of Dioscorea nipponica using successive column chromatography over silica gel, Sephadex LH-20, and preparative high performance liquid chromatography (HPLC) resulted in the isolation and identification of 17 phenolic derivatives, including four new phenolic compounds (1-4). The structural elucidation of these compounds was based on spectroscopic methods, including 1D and 2D nuclear magnetic resonance (NMR) spectroscopy techniques, mass spectrometry, and optical rotation. All isolated compounds were evaluated for their effects on nerve growth factor (NGF) secretion in a C6 rat glioma cell line and nitric oxide (NO) production in lipopolysaccharide (LPS)-activated BV2 cells. The neurite outgrowth of compound 16 was further evaluated by using mouse neuroblastoma N2a cell lines. Three new stilbene derivatives, diosniponol C (1), D (2) and diosniposide A (3) and one new phenanthrene glycoside, diosniposide B (4), together with 13 known compounds were isolated from the rhizomes of Dioscorea nipponica. Of the tested compounds (1-17), phenanthrene, 3,7-dihydroxy-2,4,6-trimethoxy-phenanthrene (16) was the most potent NGF inducer, with 162.35±16.18% stimulation, and strongly reduced NO levels with an IC50 value of 19.56 μM in BV2 microglial cells. Also, it significantly increased neurite outgrowth in N2a cells. This study supports the ethnopharmacological use of Dioscorea nipponica rhizomes as traditional medicine.


Effect of substituents in chlorin e6 derivatives on the loading efficiency of the photosensitizer into the liposome membrane and their biological activity.
In this work, we incorporated the hydrophobic alkylamide and hydroxyalkylamide derivatives of chlorin e6 into the lipid bilayer of liposomes. We obtained the data on the effectiveness of incorporation of studied compounds and have determined the size of liposomes and their stability when stored in liquid form. We also investigated the bioactivity of chlorin photosensitizers and compared the photodynamic activity of studied compounds in free and liposomal forms.


Topical equol preparation improves structural and molecular skin parameters.
Equol has been shown to improve skin health and regeneration, due to its antioxidative, phytoestrogenic and epigenetic characteristics. The effects of a topical intervention on skin structure, telomere length and epigenetic markers in skin cells were analysed. Sixty-four participants were divided in four groups and three of them treated topically with the following: emulsion with Equol powder (Isoflavandiol-E-55-RS®); emulsion with microencapsulated Equol (Vesisorb® Isoflavandiol-E-55-RS®) and an emulsion with lecithin (Vesisorb® placebo). A control group of 6 volunteers did not receive any intervention. The active compound was a 0.5% equol-racemate. For 58 participants, all samples were collected. Structural analysis, molecular analysis and questionnaires were performed at the start of the study and after 8 weeks of intervention, twice a day. Structural skin parameters were analysed by Visioscan® VC 98 and Cutometer® dual MPA 580. Molecular analyses from epidermal cells collected by skin stripping of the forehead included telomere length and LINE-1 methylation, following DNA extraction, bisulfite conversion and qPCR as well as high-resolution melting curve analysis. Effects of nutrition and lifestyle habits were evaluated with a standardized food and lifestyle questionnaire. The surface analysis showed significant improvements in skin roughness, skin texture and skin smoothness after both interventions. Cutometer® dual MPA 580 measurement revealed improvement of skin firmness and elasticity parameters for both preparations. A decrease in mean LINE-1 methylation (n.s.) and telomere length (sign. P < 0.05) was observed in the sample group with age. In the treated groups, significantly longer telomeres were observed after intervention. Whether changes in telomere length reflect changes in the regulation of telomerase, epigenetic interactions or turnover of keratinocytes needs further research. Stability and availability of preparations in skin seems to be high as not many significant differences in the activity of pure or encapsulated substances were seen. The results of this study indicate that equol has beneficial effects on structural as well as molecular skin parameters and encourages further investigations to decipher the epigenetic regulation of skin ageing and interactions of equol.


Analysis of methionine enkephalin in human pituitary by multi-dimensional reversed-phase high-performance liquid chromatography, radioreceptor assay, radioimmunoassay, fast atom bombardment mass spectrometry, and mass spectrometry-mass spectrometry.
Methionine enkephalin (ME = YGGFM) was measured in five individual human post-mortem pituitaries using four different analytical methods, with the objective of comparing the molecular specificities of the methods. Radioreceptor assay (RRA) used a receptor-rich preparation from brain and [3H]etorphine as radioligand to determine ME-like receptoractivity (ME-LR). Radioimmunoassay (RIA) measured ME-like immunoreactivity (ME-LI). Pituitary samples analyzed by RRA and RIA were purified first with a high-performance liquid chromatography (HPLC) gradient on a polymer analytical column. Fast atom bombardment mass spectrometry (FAB-MS) in two different detection modes quantified ME using the protonated molecular ion MH+ of ME at 574 a.m.u. and B/E linked-field selected reaction monitoring (SRM) to monitor the specific unimolecular metastable transition that produced the unique amino acid sequence-determining tetrapeptide fragment ion YGGFA+ from the MH+ precursor ion. Both FAB-MS methods used the deuterated internal standard YGG[2H5-F]M. Samples analyzed with FAB-MS were purified first with multi-dimensional reversed-phase HPLC. The first dimension was an ODS gradient, and the second dimension was a polymer isocratic elution. The following ME amounts were measured (mean +/- standard error of the mean): ME-LR, 7.0 +/- 1.9 micrograms g-1 tissue; ME-LI, 1.8 +/- 0.7 micrograms g-1 tissue; MH+, 2.7 +/- 0.6 micrograms g-1 tissue; SRM, 3.0 +/- 0.8 micrograms g-1 tissue. The FAB SRM method provided the highest level of molecular specificity amount these four analytical methods used to measure picomole amounts of endogenous ME in a human pituitary.