Pharmacokinetic/Pharmacodynamic Evaluation of a Novel Aminomethylcycline Antibiotic, KBP-7072, in the Neutropenic Murine Pneumonia Model against Staphylococcus aureus and Streptococcus pneumoniae.
KBP-7072 is a novel aminomethylcycline antibiotic in clinical development for community-acquired pneumonia. The goal of present studies was to determine which pharmacokinetic/pharmacodynamic (PK/PD) parameter magnitude correlated with efficacy in the murine pneumonia infection model against Staphylococcus aureus and Streptococcus pneumoniae KBP-7072 pharmacokinetic measurements were performed in plasma and epithelial lining fluid (ELF) at 4-fold-increasing doses from 1 to 256 mg/kg of body weight subcutaneously. Pharmacokinetic parameters were calculated using a noncompartmental model and were linear over the dose range. Penetration into ELF ranged from 82% to 238% comparing ELF drug concentrations to plasma free drug concentrations. Twenty-four-hour dose-ranging efficacy studies were then performed in the neutropenic murine pneumonia model against 5 S. aureus (3 methicillin-resistant and 2 methicillin-susceptible) and 6 S. pneumoniae (2 Tetr and 2 Penr) strains. KBP-7072 demonstrated potent in vivo activity resulting in a 3- to 5-log10 kill in CFU burden compared to the start of therapy for all strains. The PK/PD index area under the concentration-time curve (AUC)/MIC corelated well with efficacy (R2, 0.80 to 0.89). Net stasis was achieved at plasma 24-h free drug AUC/MIC values of 1.13 and 1.41 (24-h ELF AUC/MIC values of 2.01 and 2.50) for S. aureus and S. pneumoniae, respectively. A 1-log10 kill was achieved at 24-h plasma AUC/MIC values of 2.59 and 5.67 (24-h ELF AUC/MIC values of 4.22 and 10.08) for S. aureus and S. pneumoniae, respectively. A 2-log10 kill was achieved at 24-h plasma AUC/MIC values of 7.16 and 31.14 (24-h ELF AUC/MIC values of 8.37 and 42.92) for S. aureus and S. pneumoniae, respectively. The results of these experiments will aid in the rational design of dose-finding studies for KBP-7072 in patients with community-acquired bacterial pneumonia (CAP).


Organic Photovoltaics and Bioelectrodes Providing Electrical Stimulation for PC12 Cell Differentiation and Neurite Outgrowth.
Current bioelectronic medicines for neurological therapies generally involve treatment with a bioelectronic system comprising a power supply unit and a bioelectrode device. Further integration of wireless and self-powered units is of practical importance for implantable bioelectronics. In this study, we developed biocompatible organic photovoltaics (OPVs) for serving as wireless electrical power supply units that can be operated under illumination with near-infrared (NIR) light, and organic bioelectronic interface (OBEI) electrode devices as neural stimulation electrodes. The OPV/OBEI integrated system is capable to provide electrical stimulation (ES) as a means of enhancing neuron-like PC12 cell differentiation and neurite outgrowth. For the OPV design, we prepared devices incorporating two photoactive material systems--β-carotene/N,N'-dioctyl-3,4,9,10-perylenedicarboximide (β-carotene/PTCDI-C8) and poly(3-hexylthiophene)/phenyl-C61-butyric acid methyl ester (P3HT/PCBM)--that exhibited open circuit voltages of 0.11 and 0.49 V, respectively, under NIR light LED (NLED) illumination. Then, we connected OBEI devices with different electrode gaps, incorporating biocompatible poly(hydroxymethylated-3,4-ethylenedioxythiophene), to OPVs to precisely tailor the direct current electric field conditions during the culturing of PC12 cells. This NIR light-driven OPV/OBEI system could be engineered to provide tunable control over the electric field (from 220 to 980 mV mm(-1)) to promote 64% enhancement in the neurite length, direct the neurite orientation on chips, or both. The OPV/OBEI integrated systems under NIR illumination appear to function as effective power delivery platforms that should meet the requirements for wirelessly offering medical ES to a portion of the nervous system; they might also be a key technology for the development of next-generation implantable bioelectronics.


Non-Hodgkin's lymphomas presenting as cutaneous lesions.
To analyze the spectrum of Non-Hodgkin's Lymphomas (NHL) presenting as skin lesions and to see the histological type, age, sex incidence and site predilection of different types of NHLs and to correlate our findings with western published data. A retrospective descriptive case series study. All NHLs (nodal and extra nodal) diagnosed between January 1991-December 2001 in the section of Histopathology, Department of Pathology, The Aga Khan University, Karachi were retrieved by using SNOMED coding system. Cases were reviewed and those presenting with skin involvement were selected. Further characterization of these tumours was done by morphology and Immunohistochemistry. Panel of antibodies included leukocyte common antigen (CD45), Pan B markers (CD20 and CD 79a), Pan T markers(UCHL1), Ki-1(CD30) and epithelial membrane antigen (EMA). The total number of NHLs (nodal and extra nodal) diagnosed in our department during the study period of 10 years were 1610. Out of these, 133 cases (8.26%) of NHLs presented with skin involvement. Out of these 44.4% (n=59) were NHLs of B cell phenotype while 23.3% (n=31) cases were of T cell phenotype. Out of 133 cases, 14(10.5%) were of mycosis fungoides (MF), and 3 (2%) each were of anaplastic large cell lymphoma (ALCL:Ki-1), lymphoma and lymphoblastic lymphoma/leukemia. In 17.3% (n=23) cases the phenotype could not be ascertained. There were 97(73%) males and 36(27%) females with a M:F ratio of 2.7:1. All types of NHLs showed a site predilection for the head and neck region except MF which presented mainly as generalized body lesions. NHLs presenting as skin lesions are more commonly of B cell phenotype followed by T cell phenotype (NOS) and mycosis fungoides. Probable reasons for the high frequency of cutaneous involvement by B-cell NHL are the advanced nature of the disease at the time of presentation and biological behavior of the tumors in our population.


Improved IL-2 immunotherapy by selective stimulation of IL-2 receptors on lymphocytes and endothelial cells.
IL-2 immunotherapy is an attractive treatment option for certain metastatic cancers. However, administration of IL-2 to patients can lead, by ill-defined mechanisms, to toxic adverse effects including severe pulmonary edema. Here, we show that IL-2-induced pulmonary edema is caused by direct interaction of IL-2 with functional IL-2 receptors (IL-2R) on lung endothelial cells in vivo. Treatment of mice with high-dose IL-2 led to efficient expansion of effector immune cells expressing high levels of IL-2Rbetagamma, including CD8(+) T cells and natural killer cells, which resulted in a considerable antitumor response against s.c. and pulmonary B16 melanoma nodules. However, high-dose IL-2 treatment also affected immune cell lineage marker-negative CD31(+) pulmonary endothelial cells via binding to functional alphabetagamma IL-2Rs, expressed at low to intermediate levels on these cells, thus causing pulmonary edema. Notably, IL-2-mediated pulmonary edema was abrogated by a blocking antibody to IL-2Ralpha (CD25), genetic disruption of CD25, or the use of IL-2Rbetagamma-directed IL-2/anti-IL-2 antibody complexes, thereby interfering with IL-2 binding to IL-2Ralphabetagamma(+) pulmonary endothelial cells. Moreover, IL-2/anti-IL-2 antibody complexes led to vigorous activation of IL-2Rbetagamma(+) effector immune cells, which generated a dramatic antitumor response. Thus, IL-2/anti-IL-2 antibody complexes might improve current strategies of IL-2-based tumor immunotherapy.


Longitudinal relation between smoking and white blood cells.
Higher white blood cell counts in smokers compared with nonsmokers have been well documented, but the longitudinal relation between changes in smoking and changes in white blood cells has not been well described. Since 1984, data have been collected semiannually by the Multicenter AIDS Cohort Study (MACS), a four-center prospective cohort study of acquired immunodeficiency syndrome (AIDS) in homosexual men. The study population includes 2,435 participants who were human immunodeficiency virus (HIV) seronegative as of September 1994 and who contributed 20,918 person-visits for this analysis. For individuals who modified their smoking behavior, changes in white blood cell counts occurred primarily during the first 6 months following changes in the amount of cigarettes smoked. Among former smokers who resumed smoking, the extent of the increase in white blood cell count depended on the number of cigarettes smoked. Specifically, increases of 241, 340, and 740 cells/microliter were observed for smokers who resumed smoking < 1, 1 to < 2, and > or = 2 packs/day, respectively. Conversely, smokers who quit smoking had a decrease of white blood cell count: -32, -629, and -1,122 cells/microliter for men who previously smoked < 1, 1 to < 2, and > or = 2 packs/day, respectively. Long-term ex-smokers, however, still had higher white blood cell counts than did never smokers. There was a high within-individual correlation of white blood cell count in persons who reported a consistent level of smoking (i.e., average correlations between two white blood cell counts 6 years apart were 0.51 for never smokers, 0.48 for ex-smokers, 0.56 for men who smoked < 1 pack/day, and 0.43 for men who smoked > or = 1 pack/day). These analyses indicate an acute effect of changes in smoking on changes in white blood cell count, a residual effect of having been a smoker, and high long-term tracking for white blood cell count.


The effect of interleukin-10 gene promoter polymorphisms on early-onset coronary artery disease.
We assessed the association between interleukin-10 (IL-10) -1082G/A and -592C/A polymorphisms, and coronary heart disease (CHD). A cross-sectional, observational study included 86 patients (mean age 43.36±4.930 years) diagnosed to have CHD and 88 healthy controls (mean age 47.07±8.135 years). IL-10 -1082G/A and -592C/A polymorphisms were analyzed using restriction fragment length polymorphism (RFLP) and agarose gel electrophoresis methods in both patient and control groups. Genotype distributions of the polymorphisms between CHD patients and controls were assessed by Chi-square analysis. The genotype distribution of the -1082 G/A polymorphism was not different in premature CHD patients (GG: 38.3%; GA: 51.1%; AA: 10.6%) and controls (GG: 43.1%; GC: 43.1%; CC: 13.8%) (p=0.57). The prevalence of the A allele at -1082G/A polymorphism was 36.6% in patients and 35.3% in controls. Both allele and genotype frequencies of -592C/A polymorphism did not also differ significantly between patients with CHD and controls. We did not observe relationships between polymorphism-specific haplotypes and adverse angiographic and clinical outcomes. We have observed a significant difference of IL-10 -592C/A allelic frequency (OR=2.00 95% CI=0.9434-4.2579) between the younger CHD patients (<45 years, Group 2) and matched controls. Our study suggests that IL-10-592C/A polymorphism may play a role in susceptibility to CHD in younger patients.


Shared decision making: trade-offs between narrower and broader conceptions.
Shared decision-making approaches, by recognizing the autonomy and responsibility of both health professionals and patients, aim for an ethical 'middle way' between 'paternalistic' and 'consumerist' models of clinical decision making. Shared decision making has been understood in various ways. In this paper, we distinguish narrow and broader conceptions of shared decision making and explore their relative strengths and weaknesses. In the first part of the paper, we construct a summary characterization of an archetypal narrow conception of shared decision making (a conception that does not coincide with any specific published model but which reflects features of a variety of models). We show the shortcomings of such a conception and highlight the need to broaden out our thinking about shared decision making if the ethical (and instrumental) goals of shared decision making are to be realized. In the second part of the paper, we acknowledge and explore the advantages and disadvantages of operating with broader conceptions of shared decision making by considering the analogies between health professional-patient relationships and familiar examples of 'open-ended' relationships (e.g. friendships). We conclude by arguing that the illustrated 'trade-offs' between narrow conceptions (which may protect patients from inappropriately paternalistic professionals but preclude important forms of professional support) and broad conceptions (which render more forms of professional support legitimate but may require skills or virtues that not all health professionals possess) suggest the need to find ways, in principle and in practice, of taking seriously both patient autonomy and autonomy-supportive professional intervention.


Pulmonary tuberculosis due to Mycobacterium microti: a study of six recent cases in France.
Human tuberculosis caused by Mycobacterium microti is rare, but its prevalence and clinical significance may have been underestimated. To the best of our knowledge, 21 cases have been reported in the literature in the last decade. We report six recent pulmonary cases caused by M. microti over a period of 5 years detected in French clinical mycobacteriology laboratories of the hospital network. Our data confirm the potential of M. microti to cause clinical illness in immunocompetent patients. M. microti grew slowly from specimens, delaying the final microbiological diagnosis. Therefore, patients with tuberculosis caused by M. microti could benefit from the use of rapid diagnostic molecular techniques directly on clinical samples. From a review of the literature and this study, a classical antituberculous therapy seems effective in treating patients with M. microti disease.


Psychostimulants in the treatment of depression : a review of the evidence.
Psychostimulants have euphoric and alerting properties that suggest their usefulness in treating depressive disorders; however, problems with tolerance and dependence with some drugs militate against their widespread therapeutic use where more acceptable licensed alternatives are available. The introduction of modafinil, a stimulant not associated with tolerance and dependence, has re-awakened interest in psychostimulants as antidepressants. The available literature, while containing somewhat inconsistent data of rather poor quality, does suggest that psychostimulants have useful antidepressant properties and are usually well tolerated. They may be useful as adjuncts to standard antidepressants in refractory depression, but have particular utility in conditions where a prompt therapeutic effect is desired and where tolerance and dependence are less of a concern. Such conditions include the treatment of depression in terminal illness and in extreme old age.Psychostimulants, although now largely discarded as treatment options for depression, deserve careful consideration as potential therapeutic agents in specific patient subgroups.


An Analysis of Noise in Multi-Bit ΣΔ Modulators with Low-Frequency Input Signals.
Digital and smart sensors are commonly implemented using multi-bit ΣΔ Modulators. Undesired signals can be present at the ADC input, such as low-frequency signals with medium or high amplitude, as a consequence of mechanical artifacts in the MEMS and/or temporary signal overload. Simulations and measurements of those sensors with such signals show temporary increments of in-band noise power. This paper investigates the factors that produce this transient performance loss. Interestingly, noise increments happen when the modulator is forced to toggle between three adjacent levels and is not correlated with the typical tonal behavior of ΣΔ Modulators. Hence, the sensor performance is sensitive to some specific input patterns even if tonal behavior is decreased by dithering the input of the ADC. Different error sources, such as the mismatch between DAC cells, loop filter linearity error, and quantization error, contribute to the observed noise increments. Our aim is to analyze each of these error sources to understand and quantify in-band noise power increments, and to desensitize the ADC from the undesired input patterns. Some estimation equations are proposed and verified through extensive simulations, by means of deterministic and stochastic methods. These equations are influenced by some modulator parameters and can be used to optimize them in order to reduce such in-band noise power increments.


HPV analysis in distinguishing second primary tumors from lung metastases in patients with head and neck squamous cell carcinoma.
For patients with head and neck squamous cell carcinoma (HNSqCC), the development of squamous cell carcinoma (SqCC) in the lung may signal a new primary or the onset of metastatic dissemination. Although the distinction influences prognosis and therapy, it may not be straightforward on histologic or clinical grounds. The human papillomavirus (HPV) is an etiologic agent for SqCCs arising from the oropharynx but not for SqCCs arising from other head and neck sites. For patients with HNSqCC who develop a lung SqCC, HPV analysis could be useful in establishing tumor relationships. High-risk HPV in situ hybridization was performed on 54 lung SqCCs from patients with a previously diagnosed HNSqCC and on 166 primary lung carcinomas from patients without a prior HNSqCC. HPV was detected in 11 of 220 (5%) cases. All HPV-positive cases were from patients with a prior oropharyngeal SqCC. For the paired oropharyngeal and lung SqCCs, HPV status was concordant in 95% of cases. Time from treatment of HPV-positive oropharyngeal carcinomas to detection of lung carcinoma ranged from 1 to 97 months (mean, 36 mo). Two HPV-positive cancers were detected in the lung 8 years after treatment of the oropharyngeal primary. Despite the long interval, E6 sequencing analysis of 1 of these paired samples confirmed that the tumors harbored the same HPV-16 variant. HPV does not seem to play a role in the development of primary lung cancer. For patients with oropharyngeal SqCC who develop lung SqCC, HPV analysis may be helpful in clarifying tumor relationships. These relationships may not be obvious on clinical grounds, as HPV-related HNSqCC may metastasize long after treatment of the primary tumor.


[Changes in tight junction protein expression and permeability of colon mucosa in rats with dextran sulfate sodium-induced inflammatory bowel disease].
To develop an experimental rat model of inflammatory bowel disease (IBD) by administration of dextran sulfate sodium (DSS), and to observe changes in the tight junction protein expression and permeability of colon mucosa. Male Sprague-Dawley (SD) rats were randomly divided into control (n=27) and IBD model groups (n=27). In the IBD model group, IBD was induced by 6-day administration of 3% DSS in water followed by 14-day administration of water only. The control group was fed with water only. Pathological changes in colon mucosae were observed on days 7, 14 and 21 after DSS administration. Colon tissue specimens were collected on day 21 for measuring myeloperoxidase (MPO) activity. The transepithelial electric resistance (TEER), transepithelial potential difference (TEPD) and short circuit current (Isc) of the specimens were measured by Ussing chamber. Real-time PCR and Western blot were used to measure the mRNA and protein expression of tight junction proteins in colon epithelia. In the IBD model group, diarrhea, hemafecia and weight loss were seen. Inflammation occurred mainly in the distal colon and was characterized by crypt abscess and inflammatory cell infiltration. The IBD model group showed significantly increased MPO activity (P<0.01), significantly decreased TEER (P<0.01) and TEPD (P<0.01), and significantly increased Isc (P<0.01) compared with the control group. No claudin 2 expression of mRNA and protein was detected in the control group, and they were expressed in the IBD model group. The expression levels of claudin 3, occludin and ZO-1 in the IBD model group were significantly decreased compared with in the control group (P<0.01). IBD rats show colonic barrier dysfunction and changes in the expression of tight junction proteins. The changes in the expression of tight junction proteins may contribute to colonic barrier dysfunction in cases of IBD in the chronic recovery stage.


Right atrium and renal sodium excretion.
Inflation of a balloon in the right atrium of 13 dogs resulted in salt and water retention not attributable to changes in GFR, RPF, filtration fraction, mean arterial pressure, left atrial pressure, or renal venous pressure. The response to right atrial balloon inflation was compared with the renal effects of constriction of the abdominal aorta above the renal arteries and constriction of the ascending aorta. Neither procedure evoked the same response as balloon inflation. Results suggest that the decrease in mean arterial pressure which accompanies balloon inflation accounts for only part of the decreased salt and water excretion. There was no significant difference between the responses of denervated and intact kidneys to right atrial balloon inflation. A binary response to stretching of the right atrium is suggested, both components apparently involving hormonal mediation.


Development and application of observable response indicators for design of an effective ozone and fine particle pollution control strategy in China.
Designing effective control policies requires efficient quantification of the nonlinear response of air pollution to emissions. However, neither the current observable indicators nor the current indicators based on response-surface modeling (RSM) can fulfill this requirement. Therefore, this study developed new observable RSM-based indicators and applied them to ambient fine particle (PM2.5) and ozone (O3) pollution control in China. The performance of these observable indicators in predicting O3 and PM2.5 chemistry was compared with that of the current RSM-based indicators. H2O2×HCHO/NO2 and total ammonia ratio, which exhibited the best performance among indicators, were proposed as new observable O3- and PM2.5-chemistry indicators, respectively. Strong correlations between RSM-based and traditional observable indicators suggested that a combination of ambient concentrations of certain chemical species can serve as an indicator to approximately quantify the response of O3 and PM2.5 to changes in precursor emissions. The observable RSM-based indicator for O3 (observable peak ratio) effectively captured the strong NOx-saturated regime in January and the NOx-limited regime in July, as well as the strong NOx-saturated regime in northern and eastern China and their key regions, including the Yangtze River Delta and Pearl River Delta. The observable RSM-based indicator for PM2.5 (observable flex ratio) also captured strong NH3-poor condition in January and NH3-rich condition in April and July, as well as NH3-rich in northern and eastern China and the Sichuan Basin. Moreover, analysis of these newly developed observable response indicators suggested that the simultaneous control of NH3 and NOx emissions produces greater benefits in provinces with higher PM2.5 exposure by up to 1.2 μg m-3 PM2.5 per 10 % NH3 reduction compared with NOx control only. Control of volatile organic compound (VOC) emissions by as much as 40 % of NOx controls is necessary to obtain the cobenefits of reducing both O3 and PM2.5 exposure at the national level when controlling NOx emissions. However, the VOC-to-NOx ratio required to maintain benefits varies significantly from 0 to 1.2 in different provinces, suggesting that a more localized control strategy should be designed for each province.


An Al/P-based frustrated Lewis pair as an efficient ambiphilic ligand: coordination of boron trihalides, rearrangement, and formation of HBX₂ complexes (X = Br, I).
The Al/P-based frustrated Lewis pair (FLP) Mes2P-C(═CH-Ph)-Al(CMe3)2 (1) reacted with boron halides BX3 (X = F, Cl, Br, I) as an ambiphilic ligand to form complexes (2-5) in which the boron atoms were coordinated to phosphorus and one of the halogen atoms to aluminum. Nonplanar five-membered heterocycles resulted that had five different ring atoms (AlCPBX). The distance of the bridging halogen atoms to the AlCPB plane increased steadily with the radius of the halogen atoms. Only the BF3 adduct showed a dynamic behavior in solution at room temperature with equivalent tert-butyl or mesityl groups in the NMR spectra, while in other cases, the rigid conformation led to the magnetic inequivalence of the substituents at Al and P with well-resolved signals for each group. The BBr3 and BI3 complexes underwent in solution at room temperature a spontaneous stereoselective rearrangement with the concomitant release of isobutene. The obtained products, Mes2P-(μ-C═CH-Ph)(μ-HBX2)-AlX(CMe3) (6 and 7) may be viewed as unique adducts of a modified new Al/P-based FLP, Mes2P-C(═CH-Ph)-AlX(CMe3) (X = Br, I), with dihalogenboranes, HBX2. The trapped boranes are either completely unknown (X = I) or unstable in the free form. Quantum-chemical calculations suggest an ionic rearrangement mechanism via the formation of a borenium cation, β-hydride elimination, and hydride transfer. The bromine migration from boron to aluminum corresponds to a formal suprafacial 1,3-sigmatropic rearrangement.


GABAergic circuit dysfunction in the Drosophila Fragile X syndrome model.
Fragile X syndrome (FXS), caused by loss of FMR1 gene function, is the most common heritable cause of intellectual disability and autism spectrum disorders. The FMR1 protein (FMRP) translational regulator mediates activity-dependent control of synapses. In addition to the metabotropic glutamate receptor (mGluR) hyperexcitation FXS theory, the GABA theory postulates that hypoinhibition is causative for disease state symptoms. Here, we use the Drosophila FXS model to assay central brain GABAergic circuitry, especially within the Mushroom Body (MB) learning center. All 3 GABAA receptor (GABAAR) subunits are reportedly downregulated in dfmr1 null brains. We demonstrate parallel downregulation of glutamic acid decarboxylase (GAD), the rate-limiting GABA synthesis enzyme, although GABAergic cell numbers appear unaffected. Mosaic analysis with a repressible cell marker (MARCM) single-cell clonal studies show that dfmr1 null GABAergic neurons innervating the MB calyx display altered architectural development, with early underdevelopment followed by later overelaboration. In addition, a new class of extra-calyx terminating GABAergic neurons is shown to include MB intrinsic α/β Kenyon Cells (KCs), revealing a novel level of MB inhibitory regulation. Functionally, dfmr1 null GABAergic neurons exhibit elevated calcium signaling and altered kinetics in response to acute depolarization. To test the role of these GABAergic changes, we attempted to pharmacologically restore GABAergic signaling and assay effects on the compromised MB-dependent olfactory learning in dfmr1 mutants, but found no improvement. Our results show that GABAergic circuit structure and function are impaired in the FXS disease state, but that correction of hypoinhibition alone is not sufficient to rescue a behavioral learning impairment.


Comparison between 2D cephalometric and 3D digital model superimpositions in patients with lateral incisor agenesis treated by canine substitution.
This study aimed to assess the reliability of measurements obtained after superimposing 3-dimensional (3D) digital models by comparing them with those obtained from lateral cephalometric radiographs in patients with lateral agenesis space closure by mesialization. Data were collected from premaxillary and postmaxillary dental casts and lateral cephalometric radiographs of 26 patients presenting lateral incisor agenesis and treated with rapid maxillary expanders and space closure by mesialization of the lateral sectors. Sagittal and vertical movements of the incisors and the maxillary molars were evaluated with lateral cephalometric radiographs and digitized 3D models superimposed on the palatal area. Paired sample t tests were used to determine if any significant difference existed between the 2 measuring techniques and between 2 different localizations of superimpositions. Cephalograms and 3D digital model measurements were statistically similar in molars and incisor movements according to anteroposterior and vertical planes. Regarding incisor movements in the anteroposterior plane, measurements derived from second ruga 3D models were significantly greater than those derived from third ruga 3D digital models. The 3D model superimposition method using the palate as a reference area is clinically reliable for assessing anteroposterior and vertical tooth movement as cephalometric superimposition in patients treated with rapid maxillary expanders and space closure by mesialization.


The willingness to pay for reducing pain and pain-related disability.
We sought to identify chronic pain patients' preferences for levels of improvement in pain-related morbidity (PRM) by measuring their willingness to pay (WTP) for reducing their pain intensity and pain-related disability. The study was a cross-sectional nonrandomized design. Participants were recruited from a tertiary multidisciplinary pain center in Canada. A computer-administered discrete-choice experiment was used to explore participants' WTP for various levels of improvement to PRM. Participants chose between two varying combination of treatments that differed in terms of their level of improvement in pain intensity, level of improvement in pain-related disability, and out-of-pocket monthly cost. The WTP to completely minimize PRM was $1428 per month. Reduction in pain intensity was valued more highly than functional improvement. For every dollar, an individual was WTP to improve his/her disability to the lowest severity (mild), he/she was WTP approximately $2 to reduce pain intensity to moderate and $3 to reduce pain intensity to mild. The potential return on investment in terms of health improvement gained was $3318 per patient visit per year. The morbidity associated with chronic pain is worth approximately $1428 for every month in the chronic pain health state. From the patient's perspective, treatment and management strategies that focus on reducing pain intensity would have the greatest impact on improving health-related quality of life. Valuing health improvement in monetary terms allows for direct monetary comparisons between the costs of chronic pain interventions and their associated health returns.


Laser smoothing of binary gratings and multilevel etched structures in fused silica.
We describe a promising approach to the processing of micro-optical components, where CO(2) laser irradiation in raster scan is used to generate localized surface melting of binary or multilevel structures on silica, fabricated by conventional reactive-ion etching. The technique is shown to provide well-controlled local smoothing of step features by viscous flow under surface tension forces, relaxing the scale length of etch steps controllably between 1 and 30 microm. Uniform treatment of extended areas is obtained by raster scanning with a power stabilized, Gaussian beam profile in the 0.5 to 1 mm diameter range. For step heights of 1 microm or less, the laser-induced relaxation is symmetric, giving softening of just the upper and lower corners at a threshold power of 4.7 W, extending to symmetric long scale relaxation at 7.9 W, with the upper limit set by the onset of significant vaporization. Some asymmetry of the relaxation is observed for 3 microm high steps. Also, undercut steps or troughs produced by photolithography and etching of a deep 64 level multistep surface are found to have a polarization-dependent distortion after laser smoothing. The laser reflow process may be useful for improving the diffraction efficiency by suppressing high orders in binary diffractive optical elements, or for converting multilevel etched structures in fused silica into smoothed refractive surfaces in, for example, custom microlens arrays.


On the Relation Between lambda Lines and Phase Separations.
It has often been assumed that the slope of the isotherm involving a pair of secondary variable vanishes along a lambda line [for example, along a lambda line in the pressure-volume plane ([unk]P/[unk]V)(T) vanishes], and therefore that the specific heat for constant extensive variable (e.g., C(V)) has the greatest possible value on the lambda line and so obeys the Buckingham-Fairbank relation. It is shown here by a heuristic theoretical argument that in (3)He-(4)He solutions ([unk]mu(4)/[unk]x(3))(T) and ([unk]mu(3)/[unk]x(3))(T) probably do not vanish and C(x3) does not have its maximum possible value, although it may become infinite when x(3) --> 0. (mu(4) and mu(3) are chemical potentials of (4)He and (3)He and x(3) is the molefraction of (3)He). Only at the tricritical point does ([unk]mu(4)/[unk]x(3))(T) finally vanish and C(x3) have a value, which cannot be exceeded without the system's becoming unstable. In the case of the transition in solid NH(4)Cl the experimental facts seem to indicate that at the higher temperatures, where the transition is of higher order, ([unk]P/[unk]V)(T) does not become zero along the transition line. A statistical thermodynamic description of tricritical points is given, and shown to accord qualitatively with the experimental results for the (3)He-(4)He solutions. There is evidence that any singular behavior at the tricritical point in (3)He-(4)He is already present along the lambda line. Finally, an analysis is made of the possible behavior of binary liquid solutions, and it is shown that a tendency of C(V) to exceed its maximum value can result in a flat top on the coexistence curve.


Lean Six Sigma Redesign of a Process for Healthcare Mandatory Education in Basic Life Support-A Pilot Study.
Healthcare staff are required to undertake mandatory training programs to ensure they maintain key clinical competencies. This study was conducted in a private hospital in Ireland, where the processes for accessing mandatory training were found to be highly complex and non-user friendly, resulting in missed training opportunities, specific training license expiration, and underutilized training slots which resulted in lost time for both the trainers and trainees. A pilot study was undertaken to review the process for accessing mandatory training with a focus on the mandatory training program of Basic Life Support (BLS). This was chosen due to its importance in patient resuscitation and its requirement in the hospital achieving Joint Commission International (JCI) accreditation. A pre- and post-team-based intervention design was used with Lean Six Sigma (LSS) methodology employed to redesign the process of booking, scheduling, and delivery of BLS training leading to staff individual BLS certification for a period of two years. The redesign of the BLS training program resulted in a new blended delivery method, and the initiation of a pilot project led to a 50% increase in the volume of BLS classes and a time saving of 154 h 30 min for staff and 48 h 14 min for BLS instructors. The success of the BLS process access pilot has functioned as a platform for the redesign of other mandatory education programs and will be of interest to hospitals with mandatory training requirements that are already facing healthcare challenges and demands on staff time.


Local production of B lymphocyte stimulator protein and APRIL in arthritic joints of patients with inflammatory arthritis.
To determine whether synovial fluid (SF) levels and cell-surface expression of B lymphocyte stimulator (BLyS) protein and SF levels of APRIL are elevated in patients with inflammatory arthritis (IA). Same-day blood and SF samples from 89 patients with 103 knee effusions (81 knees with IA and 22 with noninflammatory arthritis [NIA]) were evaluated for BLyS protein and APRIL levels by enzyme-linked immunosorbent assay. Blood and SF mononuclear cells were double-stained for surface BLyS protein and surface CD14 (monocyte marker) and were analyzed by flow cytometry. Complete blood cell counts and SF nucleated cell counts were performed by the clinical hematology laboratory. BLyS protein levels were higher in SF than in corresponding serum samples from both IA and NIA patients. SF BLyS protein levels, but not surface expression of BLyS protein, were disproportionately elevated in IA patients. APRIL levels were higher in SF than in corresponding serum samples from most IA patients but not NIA patients. SF BLyS protein and APRIL levels correlated with each other, and each correlated with SF monocyte, lymphocyte, neutrophil, and total nucleated cell counts. Although SF and serum BLyS protein levels correlated with each other, SF and serum APRIL levels did not, suggesting that SF BLyS protein levels are more dependent upon systemic factors than are SF APRIL levels. Moreover, in 8 patients who underwent sequential arthrocenteses, changes in SF BLyS protein levels did not immutably parallel changes in SF APRIL levels, indicating their differential regulation. BLyS protein and APRIL are locally produced in inflamed joints. Their respective SF levels are differentially regulated, suggesting that they serve different functions. Together, their local production may foster survival and/or expansion of B cells that produce pathogenic autoantibodies and/or promote local T cell activation and consequent joint destruction.


Anti-smoking advice from general practitioners: is a population-based approach to advice-giving feasible?
General practitioners' (GPs') advice against smoking has a small, beneficial effect on patients' smoking. Consequently, GPs have been urged to adopt a population-based approach to advice-giving that involves discussing smoking repeatedly with the maximum possible number of smokers. This discussion paper assesses how far GPs' current clinical practice is from a population-based approach to advice-giving and finds that GPs prefer a problem-orientated approach to advising those who present with smoking-related problems. Discussion focuses on the feasibility of suggesting that GPs adopt a population-based approach instead.


Chiral inversion of 2-arylpropionic acid non-steroidal anti-inflammatory drugs--II. Racemization and hydrolysis of (R)- and (S)-ibuprofen-CoA thioesters.
The inversion of 2-arylpropionic acids (2-APAs) has become the subject of much attention. It is a unique reaction specific to this group of drugs. Inversion proceeds via stereoselective activation of the R-enantiomer to its CoA thioester whereby it is then racemized and hydrolysed to release free drug. The racemization and hydrolysis processes have been examined in this study using chemically synthesized CoA thioesters of the ibuprofen enantiomers and in vitro models employing rat liver homogenate and the mitochondrial and microsomal fractions as the source of the 'racemase' enzymes. Rat liver homogenate mediated the racemization and hydrolysis of both (R)- and (S)-ibuprofen-CoA thioesters. The rat liver mitochondrial fraction similarly mediated racemization and hydrolysis of both CoA thioesters. There was less racemase activity in the rat liver microsomal fraction and the data indicated that this fraction may contain two hydrolases which act separately on the (R)- and (S)-ibuprofen-CoA thioesters. The data are further evidence that the stereoselectivity of the CoA synthetase controls the overall stereoselectivity of inversion.


Data-driven prioritization and preclinical evaluation of therapeutic targets in glioblastoma.
Patients with glioblastoma (GBM) have a dismal prognosis, and there is an unmet need for new therapeutic options. This study aims to identify new therapeutic targets in GBM. mRNA expression data of patient-derived GBM (n = 1279) and normal brain tissue (n = 46) samples were collected from Gene Expression Omnibus and The Cancer Genome Atlas. Functional genomic mRNA profiling was applied to capture the downstream effects of genomic alterations on gene expression levels. Next, a class comparison between GBM and normal brain tissue was performed. Significantly upregulated genes in GBM were further prioritized based on (1) known interactions with antineoplastic drugs, (2) current drug development status in humans, and (3) association with biologic pathways known to be involved in GBM. Antineoplastic agents against prioritized targets were validated in vitro and in vivo. We identified 712 significantly upregulated genes in GBM compared to normal brain tissue, of which 27 have a known interaction with antineoplastic agents. Seventeen of the 27 genes, including EGFR and VEGFA, have been clinically evaluated in GBM with limited efficacy. For the remaining 10 genes, RRM2, MAPK9 (JNK2, SAPK1a), and XIAP play a role in GBM development. We demonstrated for the MAPK9 inhibitor RGB-286638 a viability loss in multiple GBM cell culture models. Although no overall survival benefit was observed in vivo, there were indications that RGB-286638 may delay tumor growth. The MAPK9 inhibitor RGB-286638 showed promising in vitro results. Furthermore, in vivo target engagement studies and combination therapies with this compound warrant further exploration.


Facing Spousal Cancer During Child-Rearing Years: Do Social Support and Hardiness Moderate the Impact of Psychological Distress on Quality of Life?
Partners of cancer patients report psychological distress and reduced quality of life. However, partners' mental health status and quality of life during child-rearing years and the influence of social support and hardiness on their well-being have not yet been studied. The aim of this study was to describe psychological distress, quality of life, social support, and hardiness of the partners facing spousal cancer during child-rearing years and investigate whether social support or hardiness moderated the relationship between psychological distress and quality of life. Cross-sectional data were collected in Norway from December 2013 to July 2015 as part of the Cancer-PEPSONE study. Five questionnaires were administered to 14 females and 21 males (n = 35). Participants reported more psychological distress and lower quality-of-life scores than other healthy Norwegian populations. Psychological distress seemed to be associated with their not being in control of their futures. Received social support moderated the effect of psychological distress on quality of life. Facing spousal cancer during child-rearing years seemed to have a substantial impact on partners' mental health and an adverse impact on their quality of life. Accordingly, these partners' self-care abilities may be reduced. Received social support may reduce the multiple burdens and consequently allow for enhancement of self-care. Interventions should aim to improve the social support provided to child-rearing partners, which may improve their quality of life. Providing adequate information about their partner's cancer illness and treatment may enhance their feelings of control, which may be beneficial for their mental health status.


Clinical application of objective perimetry using multifocal visual evoked potentials in glaucoma practice.
To evaluate the role of objective perimetry using multifocal visual evoked potentials (mVEPs) in glaucoma practice, and to assess its utility in patients with inconclusive standard automated perimetry findings. Method A retrospective case series of 436 consecutive subjects referred for glaucoma investigation who underwent testing with the AccuMap V1.3 mVEP system (ObjectiVision Pty, Ltd, Sydney, New South Wales, Australia) within a defined 12-month period. Sensitivity was determined by comparing this testing with that of standard automated perimetry and that used in a subgroup in whom masked stereoscopic optic disc photographs were used as an alternative reference standard. Overall clinical diagnostic outcomes were assessed. The mVEP changes were correlated with the stage of disease and Humphrey mean deviation (r = 0.78). The overall sensitivity for detecting glaucoma with established subjective field loss was 97.5% (early glaucoma, 95.0%), whereas 92.2% of low-risk suspects had normal mVEPs. When masked disc assessment alone was used for diagnosis of abnormality, sensitivity of mVEP (80.6%) and Humphrey visual field results (81.9%) were similar, but mVEP specificity was better (89.2% vs 79.5%). The mVEP was particularly useful in assessing excessive subjective field loss (45 eyes) by showing a much closer correlation with the clinical picture. Multifocal VEP is an effective method for detecting visual field loss in glaucoma. It provides a valuable aid to the clinician in categorizing patients with unreliable, variable, unconfirmed, or excessive subjective field loss.


Continuing professional education for general practitioners on chronic obstructive pulmonary disease: feasibility of a blended learning approach in Bangladesh.
Continuing medical education (CME) is essential to developing and maintaining high quality primary care. Traditionally, CME is delivered face-to-face, but due to geographical distances, and pressure of work in Bangladesh, general practitioners (GPs) are unable to relocate for several days to attend training. Using chronic obstructive pulmonary disease (COPD) as an exemplar, we aimed to assess the feasibility of blended learning (combination of face-to-face and online) for GPs, and explore trainees' and trainers' perspectives towards the blended learning approach. We used a mixed-methods design. We trained 49 GPs in two groups via blended (n = 25) and traditional face-to-face approach (n = 24) and assessed their post-course knowledge and skills. The COPD Physician Practice Assessment Questionnaire (COPD-PPAQ) was administered before and one-month post-course. Verbatim transcriptions of focus group discussions with 18 course attendees and interviews with three course trainers were translated into English and analysed thematically. Forty GPs completed the course (Blended: 19; Traditional: 21). The knowledge and skills post course, and the improvement in self-reported adherence to COPD guidelines was similar in both groups. Most participants preferred blended learning as it was more convenient than taking time out of their busy work life, and for many the online learning optimised the benefits of the subsequent face-to-face sessions. Suggested improvements included online interactivity with tutors, improved user friendliness of the e-learning platform, and timing face-to-face classes over weekends to avoid time-out of practice. Quality improvement requires a multifaceted approach, but adequate knowledge and skills are core components. Blended learning is feasible and, with a few caveats, is an acceptable option to GPs in Bangladesh. This is timely, given that online learning with limited face-to-face contact is likely to become the norm in the on-going COVID-19 pandemic.


Association between the CTLA-4 +49 A/G polymorphism and susceptibility to rheumatoid arthritis: a meta-analysis.
The aim of this study was to explore whether the cytotoxic T lymphocyte antigen-4 (CTLA-4) +49 A/G polymorphism confers susceptibility to rheumatoid arthritis (RA). A meta-analysis was conducted on the associations between CTLA-4 +49 A/G polymorphism and RA using; 1) allele contrast, 2) the recessive model, 3) the dominant model, and 4) an additive model. A total of 19 studies, 5,752 RA patients and 5,508 controls, encompassing 9 Caucasian, 8 Asian, 1 Mexican, and 1 Tunisian population were included in this meta-analysis. Ethnicity-specific meta-analysis was performed on Caucasian and Asian populations. Meta-analysis of the CTLA-4 +49 A/G polymorphism revealed an association between RA and the CTLA-4 +49 G allele in all 11,260 study subjects (odds ratio (OR) 1.118, 95% confidence interval (CI) 1.033-1.210, P = 0.005). Stratification by ethnicity showed an association between the CTLA-4 +49 G allele and RA in Asians (OR 1.164, 95% CI 1.056-1.283, P = 0.002), but no evidence of an association in Caucasians (OR 1.085, 95% CI 0.973-1.209, P = 0.431). Furthermore, associations were found between RA and the CTLA-4 +49 A/G polymorphism in Asians using the dominant and additive models, but not using the recessive model. On the other hand, no association was found between RA and the CTLA-4 +49 A/G polymorphism using the recessive, dominant, or additive models in Caucasians. This meta-analysis demonstrates that the CTLA-4 +49 A/G polymorphism confers susceptibility to RA in Asians, but not in Caucasians.


Clinicopathologic and Molecular Characteristics of Gastrointestinal MiNENs.
A mixed neuroendocrine-non-neuroendocrine neoplasm (MiNEN) is a recently defined entity that comprises a neuroendocrine tumor (NEN) component and a non-neuroendocrine tumor (nNEN) component. As MiNEN is a recently defined entity, its molecular nature is not well known. Here, we evaluated the clinicopathologic and molecular characteristics of gastrointestinal (GI) MiNENs. We performed a genomic analysis of 31 samples from 12 GI MiNEN cases using next-generation sequencing. We examined the primary NEN and nNEN components, as well as the metastatic NENs and nNENs. The relationships between the clinical tumor features (component, location, and grade) and their molecular characteristics were examined. The 12 MiNENs included in the study were found in the stomach (n=10), distal rectum (n=1), and anus (n=1). Primary MiNENs that had NENs as the major component showed a worse clinical outcome than those that had nNENs as the major component. All distant metastatic tumors originating from MiNENs were NENs. In addition, NENs generally carried 1.5 times more gene mutations and copy number variations than nNENs. The ATRX gene deletion and TP53 gene mutation were the most common variants in both components of GI MiNENs. We have revealed the detailed clinicopathologic and molecular findings with distinguishable alterations of GI MiNENs. To our knowledge, this is the first study to report the ATRX gene deletion in GI MiNENs. The molecular characteristics of GI MiNENs could provide clues to the pathogenic origin and progression of GI MiNENs.


Implementation of a 'lean' cytopathology service: towards routine same-day reporting.
To systematically assess the effects of a Lean management intervention in an academic cytopathology service. We monitored outcomes including specimen turnaround times during stepwise implementation of a lean cytopathology workflow for gynaecological and non-gynaecological cytology. The intervention resulted in a major reduction of turnaround times for both gynaecological (3rd quartile 4.1 vs 2.3 working days) and non-gynaecological cytology (3rd quartile 1.9 vs. 1.2 working days). Introduction of fully electronic reporting had additional effect over continuous staining of slides alone. The rate of non-gynaecological specimens reported the same day increased from 4.5% to 56.5% of specimens received before noon. Lean management principles provide a useful framework for organization of a cytopathology workflow. Stepwise implementation beginning with a simplified gynaecological cytology workflow allowed involved staff to monitor the effects of individual changes and allowed for a smooth transition.


Genetics of O-antigen biosynthesis in Pseudomonas aeruginosa.
Pathogenic bacteria produce an elaborate assortment of extracellular and cell-associated bacterial products that enable colonization and establishment of infection within a host. Lipopolysaccharide (LPS) molecules are cell surface factors that are typically known for their protective role against serum-mediated lysis and their endotoxic properties. The most heterogeneous portion of LPS is the O antigen or O polysaccharide, and it is this region which confers serum resistance to the organism. Pseudomonas aeruginosa is capable of concomitantly synthesizing two types of LPS referred to as A band and B band. The A-band LPS contains a conserved O polysaccharide region composed of D-rhamnose (homopolymer), while the B-band O-antigen (heteropolymer) structure varies among the 20 O serotypes of P. aeruginosa. The genes coding for the enzymes that direct the synthesis of these two O antigens are organized into two separate clusters situated at different chromosomal locations. In this review, we summarize the organization of these two gene clusters to discuss how A-band and B-band O antigens are synthesized and assembled by dedicated enzymes. Examples of unique proteins required for both A-band and B-band O-antigen synthesis and for the synthesis of both LPS and alginate are discussed. The recent identification of additional genes within the P. aeruginosa genome that are homologous to those in the A-band and B-band gene clusters are intriguing since some are able to influence O-antigen synthesis. These studies demonstrate that P. aeruginosa represents a unique model system, allowing studies of heteropolymeric and homopolymeric O-antigen synthesis, as well as permitting an examination of the interrelationship of the synthesis of LPS molecules and other virulence determinants.


Active metabolites in antiepileptic therapy. Reflections on a pharmacological joker.
Treatment with drugs that produce active metabolites in the organism may sometimes resemble playing cards that includes a joker. Both kinetically and dynamically the metabolites may be different from their parent compound, and antiepileptic drugs are no exception. Primidone, carbamazepine and valproate all have active metabolites, and the various problems which these mean for the treatment of epileptic patients are briefly discussed. Models for studying a parent compound and its active metabolite parallelly in man have been proposed for diazepam. These and other may be applied to antiepileptic drugs in order to increase the quality of their clinical use and the development of new drugs.


Consecutive 5'- and 3'-amide linkages stabilise antisense oligonucleotides and elicit an efficient RNase H response.
Antisense oligonucleotides are now entering the clinic for hard-to-treat diseases. New chemical modifications are urgently required to enhance their drug-like properties. We combine amide coupling with standard oligonucleotide synthesis to assemble backbone chimera gapmers that trigger an efficient RNase H response while improving serum life time and cellular uptake.


Notch Signaling-Induced Oscillatory Gene Expression May Drive Neurogenesis in the Developing Retina.
After integrating classic and cutting-edge research, we proposed a unified model that attempts to explain the key steps of mammalian retinal neurogenesis. We proposed that the Notch signaling-induced lateral inhibition mechanism promotes oscillatory expression of Hes1. Oscillating Hes1 inhibitory activity as a result leads to oscillatory expression of Notch signaling inhibitors, activators/inhibitors of retinal neuronal phenotypes, and cell cycle-promoting genes all within a retinal progenitor cell (RPC). We provided a mechanism explaining not only how oscillatory expression prevents the progenitor-to-precursor transition, but also how this transition happens. Our proposal of the mechanism posits that the levels of the above factors not only oscillate but also rise (with the exception of Hes1) as the factors accumulate within a progenitor. Depending on which factors accumulate fastest and reach the required supra-threshold levels (cell cycle activators or Notch signaling inhibitors), the progenitor either proliferates or begins to differentiate without any further proliferation when Notch signaling ceases. Thus, oscillatory gene expression may regulate an RPC's decision to proliferate or differentiate. Meanwhile, a post-mitotic precursor's selection of one retinal neuronal phenotype over many others depends on the expression level of key transcription factors (activators) required for each of these retinal neuronal phenotypes. Because the events described above are stochastic due to oscillatory gene expression and gene product inheritance from a mother RPC after its division, an RPC or precursor's decision requires the assignment of probabilities to specific outcomes in the selection process. While low and sustained (non-oscillatory) Notch signaling activity is required to promote the transition of retinal progenitors into various retinal neuronal phenotypes, we propose that the lateral inhibition mechanism, combined with high expression of the BMP signaling-induced Inhibitor of Differentiation (ID) protein family, promotes high and sustained (non-oscillatory) Hes1 and Hes5 expression. These events facilitate the transition of an RPC into the Müller glia (MG) phenotype at the late stage of retinal development.


Utility of Novel Rotational Load-Velocity Profiling Methods in Collegiate Softball Players.
Herring, CH, Beyer, KS, Redd, MJ, Stout, JR, and Fukuda, DH. Utility of novel rotational load-velocity profiling methods in collegiate softball players. J Strength Cond Res 38(1): 136-145, 2024-The purpose of this study was to determine the reliability of bat swing (BS) and rotational medicine ball throw (RMBT) load-velocity profiling (LVP) methods and explore relationships with batting performance in NCAA Division I softball players. Bat velocity was tracked with a swing sensor during the BS method, whereas an inertial measurement unit (IMU) tracked forearm velocity during the BS and RMBT methods. Intraclass correlation coefficients (ICC) were used for relative reliability, and coefficient of variation (CV) was used for absolute reliability. With the exception of theoretical maximum velocity (V0) using the average of top 2 peak velocities (PVavg) during the RMBT, no LVP variables were found to be reliable during the RMBT or BS method using the IMU (ICC ≤0.7; CV ≥15%). For the BS method with the swing sensor, all bat loads and V0 had acceptable reliability using peak velocity (PV) and PVavg (ICC >0.7; CV <15%), whereas all LVP variables were highly related between the multiple-load and two-load models when using PV and PVavg (r = 0.915-0.988; p < 0.01). There were significant relationships (r = 0.603-0.671; p < 0.05) between PV using the 0.99 Kg bat load and V0, and several in-game batting statistics. Practitioners may use the BS with the swing sensor as a rotational LVP assessment, although they should be cautious of aiming to improve batting performance in collegiate softball players based on the correlations reported until further research is performed.


Good reasons to leave home: proximate dispersal cues in a social spider.
Natal dispersal is a successful tactic under a range of conditions in spite of significant costs. Habitat quality is a frequent proximate cause of dispersal, and studies have shown that dispersal increases both when natal habitat quality is good or poor. In social species kin competition, favouring dispersal may be balanced by the benefits of group living, favouring philopatry. We investigated the effect of changes in the local environment on natal dispersal of adult females in a social spider species, Stegodyphus dumicola (Araneae, Eresidae), with a flexible breeding system, where females can breed either within the colony or individually following dispersal. We manipulated foraging opportunities in colonies by either removing the capture webs or by adding prey and recorded the number of dispersing females around each focal colony, and their survival and reproductive success. We predicted that increasing kin competition should increase dispersal of less-competitive individuals, while reducing competition could cause either less dispersal (less competition) or more dispersal (a cue indicating better chances to establish a new colony). Dispersal occurred earlier and at a higher rate in both food-augmented and web-removal colonies than in control colonies. Fewer dispersing females survived and reproduced in the web-removal group than in the control or food-augmented groups. The results support our prediction that worsening conditions in web-removal colonies favour dispersal, whereby increased kin competition and increased energy expenditure on web renewal cause females to leave the natal colony. By contrast, prey augmentation may serve as a habitat-quality cue; when the surrounding habitat is expected to be of high quality, females assess the potential benefit of establishing a new colony to be greater than the costs of dispersal.


Dialyzability of iron, zinc, and copper of different types of infant formulas marketed in Spain.
The bioavailability of trace elements in infant formulas is affected by different physiological and dietetic factors. In vitro methods based on element dialyzability have been proposed to estimate the bioavailability. Infant formulas of the same type but from different manufacturers can differ in the salt used for supplementation and in the contents of other components that can affect mineral bioavailability. The aim of our study is to estimate the dialyzability of iron, zinc, and copper of formulas marketed in Spain, in order to detect possible differences in formulas of the same type coming from different manufacturers. At the same time, the effects of the type of formula, the composition of the protein fraction, and the mineral content on the element dialyzability are also studied. Differences are found in the dialysis percentages of the elements studied in formulas of the same type but from different manufacturers. The formulas giving the highest dialysis percentages for the three considered elements are the hypoallergenic ones based on protein hydrolysates. No differences are observed in formulas having whey or casein as the main protein fraction. Significant correlations are obtained between the element contents and the dialyzability of the elements.


2HiGWAS: a unifying high-dimensional platform to infer the global genetic architecture of trait development.
Whole-genome search of genes is an essential approach to dissecting complex traits, but a marginal one-single-nucleotide polymorphism (SNP)/one-phenotype regression analysis widely used in current genome-wide association studies fails to estimate the net and cumulative effects of SNPs and reveal the developmental pattern of interplay between genes and traits. Here we describe a computational framework, which we refer to as two-side high-dimensional genome-wide association studies (2HiGWAS), to associate an ultrahigh dimension of SNPs with a high dimension of developmental trajectories measured across time and space. The model is implemented with a dual dimension-reduction procedure for both predictors and responses to select a sparse but full set of significant loci from an extremely large pool of SNPs and estimate their net time-varying effects on trait development. The model can not only help geneticists to precisely identify an entire set of genes underlying complex traits but also allow them to elucidate a global picture of how genes control developmental and dynamic processes of trait formation. We investigated the statistical properties of the model via extensive simulation studies. With the increasing availability of GWAS in various organisms, 2HiGWAS will have important implications for genetic studies of developmental compelx traits.


Approach to patients with unexplained multimorbidity with sensitivities.
To explore the underlying causation of unexplained multimorbidity with sensitivities and to discuss the management of patients who present with this perplexing condition. Medical and scientific literature was used from MEDLINE (PubMed), several books, toxicology and allergy journals, conference proceedings, government publications, and environmental health periodicals. Multimorbidity with sensitivities has become an increasingly common and confusing primary care dilemma. Escalating numbers of debilitated individuals are now presenting to family physicians and specialists with multisystem health complaints, including sensitivities and fatigue, with no obvious causation, a paucity of laboratory findings, and a lack of straightforward solutions. In the recent scientific literature, there is discussion of sensitivity-related illness, an immune-mediated disorder that frequently manifests with multisystem symptoms, commonly including sensitivities and fatigue. This condition appears to be originally caused by adverse environmental exposures and toxicant bioaccumulation-an increasingly prevalent and well-documented problem in contemporary culture. Various toxic exposures and their bioaccumulation within the body frequently manifest as sensitivity-related illness. In clinical settings, patients with this disorder often present with otherwise unexplained multimorbidity and sensitivities. The health status of patients with this condition can be ameliorated by removing triggers (eg, scented products), optimizing biochemistry, removing further sources of toxicant exposures, and eliminating the internal dose of persistent toxicants.


Pathogenic Roles of Glutamic Acid Decarboxylase 65 Autoantibodies in Cerebellar Ataxias.
Reports suggesting a pathogenic role of autoantibodies directed against glutamic acid decarboxylase 65 (GAD65Abs) in cerebellar ataxias (CAs) are reviewed, and debatable issues such as internalization of antibodies by neurons and roles of epitopes are discussed. GAD65 is one of two enzymes that catalyze the conversion of glutamate to the inhibitory neurotransmitter gamma-aminobutyric acid (GABA). A pathogenic role of GAD65Ab in CAs is suggested by in vivo and in vitro studies. (1) Intracerebellar administration of cerebrospinal fluid (CSF) immunoglobulins (IgGs) obtained from GAD65Ab-positive CA patients impairs cerebellar modulation of motor control in rats. (2) CSF IgGs act on terminals of GABAergic neurons and decrease the release of GABA in cerebellar slices from rats and mice. (3) Absorption of GAD65Ab by recombinant GAD65 diminishes the above effects, and monoclonal human GAD65Ab (b78) mimic the effects of CSF IgGs in vivo and in vitro. Studies using GAD65-KO mice confirm that the target molecule is GAD65. (4) Notably, the effects of GAD65Ab depend on the epitope specificity of the monoclonal GAD65Ab. Taken together, these results indicate that epitope-specific GAD65Ab-induced impairment of GABA release is involved in the pathogenesis of GAD65Ab-positive CA and support the early detection of GAD65Ab-associated CA to initiate immunotherapy before irreversible neuronal death in the cerebellum.


Cognitive Remediation Therapy Does Not Enhance Treatment Effect in Obsessive-Compulsive Disorder and Anorexia Nervosa: A Randomized Controlled Trial.
Guideline-recommended therapies are moderately successful in the treatment of obsessive-compulsive disorder (OCD) and anorexia nervosa (AN), leaving room for improvement. Cognitive inflexibility, a common trait in both disorders, is likely to prevent patients from engaging in treatment and from fully benefiting from existing therapies. Cognitive remediation therapy (CRT) is a practical augmentation intervention aimed at ameliorating this impairing cognitive style prior to disorder-specific therapy. To compare the effectiveness of CRT and a control treatment that was not aimed at enhancing flexibility, named specialized attention therapy (SAT), as add-ons to treatment as usual (TAU). In a randomized controlled multicenter clinical trial, 71 adult patients with OCD and 61 with AN were randomized to ten twice-weekly sessions with either CRT or SAT, followed by TAU. Patients were evaluated at baseline, post-CRT/SAT, and after 6 and 12 months, with outcomes being quantified using the Yale-Brown Obsessive Compulsive Scale for OCD and the Eating Disorder Examination Questionnaire for AN. Across study groups, most importantly CRT+TAU was not superior to control treatment (SAT)+TAU in reducing OCD and AN pathology. Contrary to expectations, SAT+TAU may have been more effective than CRT+TAU in patients being treated for OCD. CRT did not enhance the effect of TAU for OCD and AN more than SAT. Unexpectedly, SAT, the control condition, may have had an augmentation effect on TAU in OCD patients. Although this latter finding may have been due to chance, the effect of SAT delivered as a pretreatment add-on intervention for adults with OCD and AN merits future efforts at replication.


Interference of gut-brain-gonad axis originating from triclocarban exposure to parent zebrafish induces offspring embryonic development abnormality by up-regulation of maternal circSGOL1.
Triclocarban (TCC) is a widely used antibacterial ingredient possessing acute toxicity effects; however, its chronic toxicity and underlying molecular mechanisms remain uncertain. Herein, we demonstrated that chronic TCC exposure affects the growth and development of adult zebrafish through inducing an intestinal flora disorder in the gut. The imbalance of intestinal flora caused functional barriers within the intestinal-brain-gonadal axis. This resulted in a series of anomalous nerve and motor behaviors, and reproductive toxicity as reflected in pathological damage to parental gonads and F1-larval developmental malformations. Abnormal development of F1 larvae was attributed to apoptosis induced by the up-regulation of circSGOL1. This up-regulation affected the activity and localization of the hnRNP A1 protein, which then promoted overexpression of pro-apoptotic related genes that ultimately lead to apoptosis during early embryonic development. Overall, these novel findings systematically elucidated the TCC toxicity mechanism in parent-offspring dyads, and provide important theoretical guidance for early risk warning and control of chronic TCC toxicity.


Cholesterol-loading of membranes of normal erythrocytes inhibits phospholipid repair and arachidonoyl-CoA:1-palmitoyl-sn-glycero-3-phosphocholine acyl transferase. A model of spur cell anemia.
Spur cell anemia may occur in severe liver disease including alcoholic cirrhosis. Spur cell anemia red blood cells (RBCs) have a characteristic morphology, with irregular projections, an increased ratio of membrane cholesterol (Ch) to phospholipid, evidence of oxidative damage, and shortened survival resulting in hemolytic anemia. Normal RBCs may acquire many of the features of spur cells either by transfusion into a spur cell patient or in an in vitro model system that loads the RBC membrane with Ch relative to phospholipid by means of Ch-rich, phospholipid-Ch sonicates. We found evidence of abnormal phospholipid repair metabolism in spur cell anemia RBCs characterized by decreased arachidonate (Ar) uptake into phospholipids and by increased uptake into a fatty acid membrane repair intermediate, acylcarnitine (AcylCn). To study the possible modulation of phospholipid repair metabolism in spur cells by Ch-loading, we compared the Ar metabolism of RBCs loaded with Ch in vitro with that of control cells incubated in autologous serum. Ar, a polyunsaturated fatty acid, is especially sensitive to peroxidation and, thus, is likely to be involved in phospholipid repair. Ch-loading decreased the incorporation of [14C]Ar into total lipids (Ch-loaded, 1,113 +/- 48 pmol/10(10) RBCs; control, 1,525 +/- 48 pmol/10(10) RBCs) including phosphatidylethanolamine, phosphatidylserine, and phosphatidylcholine. Uptake of [14C]Ar into AcylCn increased (control AcylCn, 169 +/- 31 pmol/10(10) RBCs; Ch-loaded AcylCn, 196 +/- 35 pmol/10(10) RBCs; P = .0012). Thimerosal, an inhibitor of arachidonoyl- CoA:l-palmitoyl-sn-glycero-3-phosphocholine acyl transferase or lysophosphocholine acyl transferase (LAT), produced a similar pattern of metabolic abnormality, with decreased incorporation into phospholipid but relative increase into AcylCn. We assayed LAT in RBC membranes from Ch-loaded RBCs, using [14C]arachidonoyl CoA as precursor, and found similar decreased LAT activity at concentrations of 1-palmitoyllysophosphatidylcholine (LPC) from 1 to 30 micromol/L. Similar LAT assay results were obtained using [14C]palmitoyl LPC as the precursor. We conclude that Ch-loading of RBC membranes results in inhibition of LAT in the cell-free system in vitro and may account for the inhibited phospholipid repair in Ch-loaded intact RBCs in vitro and in spur cell anemia RBCs in vivo. Decreased ability to replace peroxidized membrane fatty acid by this metabolic pathway may contribute to the hemolytic process in spur cell anemia.


Iron Efflux by PmtA Is Critical for Oxidative Stress Resistance and Contributes Significantly to Group A Streptococcus Virulence.
Group A Streptococcus (GAS) is a human-only pathogen that causes a spectrum of disease conditions. Given its survival in inflamed lesions, the ability to sense and overcome oxidative stress is critical for GAS pathogenesis. PerR senses oxidative stress and coordinates the regulation of genes involved in GAS antioxidant defenses. In this study, we investigated the role of PerR-controlled metal transporter A (PmtA) in GAS pathogenesis. Previously, PmtA was implicated in GAS antioxidant defenses and suggested to protect against zinc toxicity. Here, we report that PmtA is a P1B4-type ATPase that functions as an Fe(II) exporter and aids GAS defenses against iron intoxication and oxidative stress. The expression of pmtA is specifically induced by excess iron, and this induction requires PerR. Furthermore, a pmtA mutant exhibited increased sensitivity to iron toxicity and oxidative stress due to an elevated intracellular accumulation of iron. RNA-sequencing analysis revealed that GAS undergoes significant alterations in gene expression to adapt to iron toxicity. Finally, using two mouse models of invasive infection, we demonstrated that iron efflux by PmtA is critical for bacterial survival during infection and GAS virulence. Together, these data demonstrate that PmtA is a key component of GAS antioxidant defenses and contributes significantly to GAS virulence.


Heterogeneity of neoplastic stem cells: theoretical, functional, and clinical implications.
Accumulating evidence suggests that human cancers develop through a step-wise, but nonlinear process of cellular diversification and evolution. Recent mutational analyses indicate that this process is more complex and diverse than anticipated before whole-genome sequencing methods were readily available. Examples are also emerging now of genetically abnormal clones of cells that have acquired mutations with known oncogenic potential but, nevertheless, may show no manifestations of malignant change for many years. To accommodate these diverse realities, we suggest the term neoplastic refer to clones of cells that have any type of somatic aberrancy associated with an increased propensity to become malignant, and the derivative term neoplastic stem cell be adopted to identify the cells responsible for the long-term maintenance of such clones. Neoplastic clones would thus include those that never evolve further, as well as those that eventually give rise to fully malignant populations, and all stages in between. The term cancer stem cells would then be more appropriately restricted to cells generating subclones that have established malignant properties. More precise molecular understanding of the different stem cell states thus distinguished should contribute to the development of more effective prognostic and therapeutic tools for cancer diagnosis and treatment.


Analysis of vascular mechanical properties of the yellow anaconda reveals increased elasticity and distensibility of the pulmonary artery during digestion.
In animals with functional division of blood systemic and pulmonary pressures, such as mammals, birds, crocodilians and a few non-crocodilian reptiles, the vessel walls of systemic and pulmonary arteries are exquisitely adapted to endure different pressures during the cardiac cycle, systemic arteries being stronger and stiffer than pulmonary arteries. However, the typical non-crocodilian reptile heart possesses an undivided ventricle that provides similar systolic blood pressure to both circuits. This raises the question whether in these species the systemic and pulmonary mechanical vascular properties are similar. Snakes also display large organ plasticity and increased cardiac output in response to digestion, and we speculate how the vascular circuit would respond to this further stress. We addressed these questions by testing the mechanical vascular properties of the dorsal aorta and the right pulmonary artery of fasted and fed yellow anacondas, Eunectes notaeus, a snake without functional ventricular separation that also exhibits large metabolic and cardiovascular responses to digestion. Similar to previous studies, the dorsal aorta was thicker, stronger, stiffer and more elastic than the pulmonary artery. However, unlike any other species studied so far, the vascular distensibility (i.e. the relative volume change given a pressure change) was similar for the two circuits. Most striking, the pulmonary artery elasticity (i.e. its capacity to resume its original form after being stretched) and distensibility increased during digestion, which suggests that this circuit is remodeled to accommodate the larger stroke volume and enhance the Windkessel effect, thus providing a more constant blood perfusion during digestion.


Secondary tics and tourettism.
Motor and phonic tics are most frequently due to Tourette syndrome, but there are many other causes of tics. We analyzed data on 155 patients with tics and co-existent disorders (101M/54F; mean age 40.5 +/- 20.2 years). Fourteen (9.0%) patients had tics associated with an insult to the basal ganglia, such as head trauma (N = 4, 2.5%), stroke (N = 2, 1.2%), encephalitis (N = 3, 1.9%) and other causes. In addition, certain drugs, toxins, and post-infectious causes were associated with tics. Rarely, peripheral injury can cause movement disorders, including tics (N = 1, 0.6%). Pervasive developmental disorders, including Asperger's syndrome (N = 13, 8.3%), mental retardation (N = 4, 2.5%), autism (N = 3, 1.9%), and Savant's syndrome (N = 1, 0.6%), also may be associated with tics, as noted in 21 of the 155 patients (13.5%). Genetic and chromosomal disorders, such as Down's syndrome 5 (3.2%), neuroacanthocytosis (N = 2, 1.2%), and Huntington's disease (N = 1, 0.6%), were associated with tics in 16 patients (10.3%). We have also examined the co-existence of tics and other movement disorders such as dystonia (N = 31, 20.0%) and essential tremor (N = 17, 10.9%). Sixteen (10.3%) patients presented psychogenic tics, and one (0.6%) psychogenic tics and dystonia; conversely, Tourette syndrome preceded the onset of psychogenic dystonia (N = 1, 0.6%), and psychogenic tremor (N = 1, 0.6%) in two patients. Finally, 12 (7.7%) patients had tics in association with non-movement related neurological disorders, such as static encephalopathy (N = 2, 1.2%) and seizures (N = 3, 1.9%). To understand the physiopathology of tics and Tourette syndrome, it is important to recognize that these may be caused or associated with other disorders.


Higher expression of the androgen-regulated gene PSA/HK3 mRNA in prostate cancer tissues predicts biochemical recurrence-free survival.
Alterations of the androgen receptor (AR)-mediated signaling through numerous mechanisms are increasingly recognized in prostate cancer (CaP) progression. We hypothesized that the assessment of well-defined AR transcriptional targets (e.g., PSA/HK3 mRNA) in CaP tissues will provide in vivo readout of AR dysfunctions. Moreover, quantitative expression features of PSA/HK3 mRNA in prostate tumor cells may serve as a prognostic indicator of disease progression. Paired benign and malignant epithelial cells (242 specimens) were obtained from laser capture microdissection of frozen OCT-embedded tissue sections prepared from radical prostatectomy specimens of 121 patients. Quantitative expression of PSA/HK3 mRNA in the matched malignant and benign cells was analyzed by real-time reverse transcription-PCR. CaP cells express significantly lower PSA/HK3 mRNA levels than matched benign cells (P = 0.0133). Moreover, low PSA/HK3 mRNA expression in malignant cells was associated with increased risk of biochemical recurrence (P = 0.0217), as well as with time to recurrence (P = 0.0371), in patients with intermediate preoperative serum prostate-specific antigen levels (2-10 ng/mL). The expression of androgen-dependent genes in clinical samples correlates with each other in patients with higher expression of PSA/HK3 mRNA but not in patients with lower expression of PSA/HK3 mRNA reflecting AR pathway dysfunction. Our study has unraveled a novel prognostic utility of quantitative measurements of PSA/HK3 mRNA reflecting AR transcriptional activity in CaP cells, which is independent of serum prostate-specific antigen. It also has potential in stratifying subsets of patients exhibiting progressive disease associated with dampened AR transcriptional functions who may be targeted by tailored therapeutic strategies.


NCCN Guidelines® Insights: Survivorship, Version 1.2022.
The NCCN Guidelines for Survivorship are intended to help healthcare professionals who work with survivors to ensure that the survivors' complex and varied needs are addressed. The NCCN Guidelines provide screening, evaluation, and treatment recommendations for the consequences of adult-onset cancer and its treatment; recommendations to help promote physical activity, weight management, and immunizations in survivors; and a framework for care coordination. This article summarizes updates to the NCCN Guidelines pertaining to preventive health for cancer survivors, including recommendations about alcohol consumption and vaccinations.


Mutagenicity and carcinogenicity of tea, Camellia sinensis.
Aqueous, caffeine free and tannin fractions of commercial tea and tannic acid were tested for mutagenicity in Ames test. Tea fractions of tannic acid were non mutagenic in strains TA 100, TA 98, TA 1535 and TA 1538 of Salmonella typhimurium with or without metabolic activation (rat-S9 mix) at different doses tested. In strain TA 98 the above tea fractions and tannic acid inhibited the S9 mix mediated mutagenicity of tobacco in a dose dependent manner. The different tea fractions at 60 degrees C, did not increase the tumor incidence in Swiss mice by gavage feeding. They also failed to produce tumors when injected subcutaneously. Caffeine free tea extract decreased the tobacco induced liver tumors but had no effect on lung tumors. The same fraction was ineffective in hexachlorocyclohexane induced liver tumors in Swiss mice.


Effect of arginine vasopressin on CSF composition and bulk flow in hyperosmolar state.
To find out whether central arginine vasopressin (AVP) plays a role in regulation of cerebrospinal fluid (CSF) composition and of CSF bulk flow (CSFbf) in acute hyperosmolar state, the experiments were performed on 16 anesthetized cats in which the ventriculocisternal system was perfused either with artificial CSF (aCSF; 8 control experiments) or with aCSF containing AVP (10 microU/min, 8 experiments) before, during, and after intravenous infusion of 5% NaCl. In both series, hyperosmotic infusion significantly increased CSF osmolality (CSFosmol) and sodium (CSFNa) and potassium (CSFK) concentration and reduced CSFbf. Administration of AVP significantly influenced the dynamics of changes in CSFNa and slightly affected changes in CSFK concentration induced by infusion of hyperosmotic saline, while it did not modify changes in CSFosmol) and CSFbf. In control experiments, CSFbf was significantly related to plasma and CSF osmolalities and sodium concentrations, whereas in AVP experiments these relationships were markedly suppressed. The results suggest that during acute hyperosmolar state AVP significantly influences sodium handling in CSF and may play a role in adaptation of the brain to hyperosmolality.


Identification of metabolic pattern and bioactive form of resveratrol in human medulloblastoma cells.
Cancer preventive reagent trans-resveratrol is intracellularly biotransformed to different metabolites. However, it is still unclear whether trans-resveratrol exerts its biological effects directly or through its metabolite(s). This issue was addressed here by identifying the metabolic pattern and the bioactive form of resveratrol in a resveratrol-sensitive human medulloblastoma cell line, UW228-3. The cell lysates and condition media of UW228-3 cells with or without 100 microM resveratrol treatment were analyzed by HPLC and LC/MS which revealed (1) that resveratrol was chemically unstable and the spontaneous generation of cis-resveratrol reduced resveratrol's anti-medulloblastoma efficacy and (2) that resveratrol monosulfate was the major metabolite of the cells. To identify the bioactive form of resveratrol, a mixture-containing approximately half fraction of resveratrol monosulfate was prepared by incubating trans-resveratrol with freshly prepared rat brain lysates. Medulloblastoma cells treated by 100 microM of this mixture showed attenuated cell crisis. The overall levels of the three brain-associated sulfotransferases (SULT1A1, 1C2 and 4A1) were low in medulloblastoma cells in vivo and in vitro in comparison with that in human noncancerous and rat normal cerebella; resveratrol could more or less up-regulate the production of these enzymes in UW228-3 cells but their overall level was still lower than that in normal cerebellum tissue. Our study thus demonstrated for the first time that trans-resveratrol is the bioactive form in medulloblastoma cells in which the expression of brain-associated SULTs was down-regulated, resulting in the increased intracellular bioavailability and anti-medulloblastoma efficacy of trans-resveratrol.


A comparison of recalled date of last menstrual period with prospectively recorded dates.
Women are often asked to recall the first day of their last menstrual period (LMP date) in a clinic setting (i.e., pregnancy dating, x-rays). There are no data supporting the validity of these reports. Using data from a group of 385 women ages 35-49 from a larger cohort study in the Washington, DC, area, we constructed menstrual segments from a prospective daily menstrual record. We then compared the first day of a menstrual segment to a woman's recalled LMP date at a subsequent study-related clinic appointment to assess the accuracy of recall. More than half of the women (56%) accurately recalled their LMP date; 74% were within 1 day, and 81% were within 2 days. Women tended to underreport (25%) the length of time since their last menstrual period rather than overreport the length of time (19%). Recall accuracy did not vary significantly with education or by whether the woman usually recorded her menstrual cycle when not in the study. As one might expect, women with a shorter recall duration tended to report more accurately. Women appear to recall their LMP dates fairly accurately, but inaccurate recall was not random. When length of recall was 3 weeks or longer, women tended to overestimate the time since LMP. This suggests that gestational age calculated from LMP date will tend to be overestimated. Most women can recall the date of their LMP reasonably well regardless of their education and whether they usually record their LMP dates.


Effects of hydrostatic pressure on growth and luminescence of a moderately-piezophilic luminous bacteria Photobacterium phosphoreum ANT-2200.
Bacterial bioluminescence is commonly found in the deep sea and depends on environmental conditions. Photobacterium phosphoreum ANT-2200 has been isolated from the NW Mediterranean Sea at 2200-m depth (in situ temperature of 13°C) close to the ANTARES neutrino telescope. The effects of hydrostatic pressure on its growth and luminescence have been investigated under controlled laboratory conditions, using a specifically developed high-pressure bioluminescence system. The growth rate and the maximum population density of the strain were determined at different temperatures (from 4 to 37°C) and pressures (from 0.1 to 40 MPa), using the logistic model to define these two growth parameters. Indeed, using the growth rate only, no optimal temperature and pressure could be determined. However, when both growth rate and maximum population density were jointly taken into account, a cross coefficient was calculated. By this way, the optimum growth conditions for P. phosphoreum ANT-2200 were found to be 30°C and, 10 MPa defining this strain as mesophile and moderately piezophile. Moreover, the ratio of unsaturated vs. saturated cellular fatty acids was found higher at 22 MPa, in agreement with previously described piezophile strains. P. phosphoreum ANT-2200 also appeared to respond to high pressure by forming cell aggregates. Its maximum population density was 1.2 times higher, with a similar growth rate, than at 0.1 MPa. Strain ANT-2200 grown at 22 MPa produced 3 times more bioluminescence. The proposed approach, mimicking, as close as possible, the in situ conditions, could help studying deep-sea bacterial bioluminescence and validating hypotheses concerning its role into the carbon cycle in the deep ocean.


Crystalline lipid domains: characterization by X-ray diffraction and their relation to biology.
Biological membranes comprise thousands of different lipids, differing in their alkyl chains, headgroups, and degree of saturation. It is estimated that 5% of the genes in the human genome are responsible for regulating the lipid composition of cell membranes. Conceivably, the functional explanation for this diversity is found, at least in part, in the propensity of lipids to segregate into distinct domains, which are important for cell function. X-ray diffraction has been used increasingly to characterize the packing and phase behavior of lipids in membranes. Crystalline domains have been studied in synthetic membranes using wide- and small-angle X-ray scattering, and grazing incidence X-ray diffraction. Herein we summarize recent results obtained using the various X-ray methods, discuss the correlation between crystalline domains and liquid ordered domains studied with other techniques, and the relevance of crystalline domains to functional lipid domains in biological membranes.


[1,2,4]triazolo[4,3-a]quinoxaline as Novel Scaffold in the Imiqualines Family: Candidates with Cytotoxic Activities on Melanoma Cell Lines.
Cutaneous melanoma is one of the most aggressive human cancers and is the deadliest form of skin cancer, essentially due to metastases. Novel therapies are always required, since cutaneous melanoma develop resistance to oncogenic pathway inhibition treatment. The Imiqualine family is composed of heterocycles diversely substituted around imidazo[1,2-a]quinoxaline, imidazo[1,2-a]pyrazine, imidazo[1,5-a]quinoxaline, and pyrazolo[1,5-a]quinoxaline scaffolds, which display interesting activities on a panel of cancer cell lines, especially melanoma cell lines. We have designed and prepared novel compounds based on the [1,2,4]triazolo[4,3-a]quinoxaline scaffold through a common synthetic route, using 1-chloro-2-hydrazinoquinoxaline and an appropriate aldehyde. Cyclization is ensured by an oxidation-reduction mechanism using chloranil. The substituents on positions 1 and 8 were chosen based on previous structure-activity relationship (SAR) studies conducted within our heterocyclic Imiqualine family. Physicochemical parameters of all compounds have also been predicted. A375 melanoma cell line viability has been evaluated for 16 compounds. Among them, three novel [1,2,4]triazolo[4,3-a]quinoxalines display cytotoxic activities. Compounds 16a and 16b demonstrate relative activities in the micromolar range (respectively, 3158 nM and 3527 nM). Compound 17a shows the best EC50 of the novel series (365 nM), even if EAPB02303 remains the lead of the entire Imiqualine family (3 nM).


Using Hyperfine Electron Paramagnetic Resonance Spectroscopy to Define the Proton-Coupled Electron Transfer Reaction at Fe-S Cluster N2 in Respiratory Complex I.
Energy-transducing respiratory complex I (NADH:ubiquinone oxidoreductase) is one of the largest and most complicated enzymes in mammalian cells. Here, we used hyperfine electron paramagnetic resonance (EPR) spectroscopic methods, combined with site-directed mutagenesis, to determine the mechanism of a single proton-coupled electron transfer reaction at one of eight iron-sulfur clusters in complex I, [4Fe-4S] cluster N2. N2 is the terminal cluster of the enzyme's intramolecular electron-transfer chain and the electron donor to ubiquinone. Because of its position and pH-dependent reduction potential, N2 has long been considered a candidate for the elusive "energy-coupling" site in complex I at which energy generated by the redox reaction is used to initiate proton translocation. Here, we used hyperfine sublevel correlation (HYSCORE) spectroscopy, including relaxation-filtered hyperfine and single-matched resonance transfer (SMART) HYSCORE, to detect two weakly coupled exchangeable protons near N2. We assign the larger coupling with A(1H) = [-3.0, -3.0, 8.7] MHz to the exchangeable proton of a conserved histidine and conclude that the histidine is hydrogen-bonded to N2, tuning its reduction potential. The histidine protonation state responds to the cluster oxidation state, but the two are not coupled sufficiently strongly to catalyze a stoichiometric and efficient energy transduction reaction. We thus exclude cluster N2, despite its proton-coupled electron transfer chemistry, as the energy-coupling site in complex I. Our work demonstrates the capability of pulse EPR methods for providing detailed information on the properties of individual protons in even the most challenging of energy-converting enzymes.


Betaine attenuates LPS-induced downregulation of Occludin and Claudin-1 and restores intestinal barrier function.
The intestinal epithelial barrier, which works as the first line of defense between the luminal environment and the host, once destroyed, it will cause serious inflammation or other intestinal diseases. Tight junctions (TJs) play a vital role to maintain the integrity of the epithelial barrier. Lipopolysaccharide (LPS), one of the most important inflammatory factors will downregulate specific TJ proteins including Occludin and Claudin-1 and impair integrity of the epithelial barrier. Betaine has excellent anti-inflammatory activity but whether betaine has any effect on TJ proteins, particularly on LPS-induced dysfunction of epithelial barriers remains unknown. The purpose of this study is to explore the pharmacological effect of betaine on improving intestinal barrier function represented by TJ proteins. Intestinal porcine epithelial cells (IPEC-J2) were used as an in vitro model. The results demonstrated that betaine enhanced the expression of TJ proteins while LPS (1 μg/mL) downregulates the expression of these proteins. Furthermore, betaine attenuates LPS-induced decreases of TJ proteins both shown by Western blot (WB) and Reverse transcription-polymerase chain reaction (RT-PCR). The immunofluorescent images consistently revealed that LPS induced the disruption of TJ protein Claudin-1 and reduced its expression while betaine could reverse these alterations. Similar protective role of betaine on intestinal barrier function was observed by transepithelial electrical resistance (TEER) approach. In conclusion, our research demonstrated that betaine attenuated LPS-induced downregulation of Occludin and Claudin-1 and restored the intestinal barrier function.


Common sensory inputs and differential excitability of segmentally homologous reticulospinal neurons in the hindbrain.
In the hindbrain of zebrafish and goldfish, reticulospinal (RS) neurons are arranged in seven segments, with segmental homologs in adjacent segments. The Mauthner cell (M-cell) in the fourth segment (r4) is known to trigger fast escape behavior. Its serial homologs, MiD2cm in r5 and MiD3cm in r6, are predicted to contribute to this behavior, which can be evoked by head-tap stimuli. However, little is known about their input-output properties. Therefore, we studied afferent projections from the auditory posterior eighth nerve (pVIIIn) and firing properties of MiD2cm and MiD3cm for comparison with the M-cell in adult goldfish. Labeling of RS neurons and the pVIIIn afferents with fluorescent tracers showed that the pVIIIn projected to r4-r6. Tone burst and electrical stimulation of the pVIIIn evoked EPSPs in the M-cell, MiD2cm, and MiD3cm. Stepwise depolarization typically elicited a single spike at the onset in the M-cell but repetitive spiking in MiD2cm and MiD3cm. This atypical property of the M-cell was mediated by dendrotoxin-I (DTX-I)-sensitive voltage-gated potassium channels together with recurrent inhibition, because combined application of DTX-I, strychnine, and bicuculline led to continuous repetitive firing in M-cells. The M-cell but not MiD2cm or MiD3cm expressed Kv1.2, a DTX-I-sensitive potassium channel subunit. Thus, the M-cell and its segmental homologs may sense common auditory information but send different outputs to the spinal circuits to control adaptive escape behavior.


Primary health care in the Czech Republic: brief history and current issues.
The objective of this paper is to describe the recent history, current situation and perspectives for further development of the integrated system of primary care in the Czech Republic. The role of primary care in the whole health care system is discussed and new initiatives aimed at strengthening and integrating primary care are outlined. Changes brought about by the recent reform processes are generally seen as favourable, however, a lack of integration of health services under the current system is causing various kinds of problems. A new strategy for development of primary care in the Czech Republic encourages integration of care and defines primary care as co-ordinated and complex care provided at the level of the first contact of an individual with the health care system.