Observation and Clinical Pattern in Patients with White Dot Syndromes: The Role of Color Photography in Monitoring Ocular Changes in Long-Term Observation.
BACKGROUND The aim of this study was to assess the clinical course and distinctive features of different white dot syndromes (WDS) in patients attending the Ophthalmology Department, Medical University of Warsaw in the years 1995-2015. MATERIAL AND METHODS Sixty-two (62) patients (43 females and 19 males), aged 18 to 77 years, referred with a WDS were included in this prospective study, with observation period ranging from 5 months to 16 years. All patients underwent a complete ophthalmological examination and multimodal imaging studies. RESULTS In this cohort of 62 patients, the following WDS entities were identified: multifocal choroiditis with panuveitis (MFCPU), multifocal choroiditis (MFC), punctate inner choroidopathy (PIC), birdshot, acute posterior multifocal placoid pigment epitheliopathy (APMPPE), subretinal fibrosis and uveitis, multiple evanescent white dot syndrome (MEWDS), serpiginous choroiditis, and single cases of acute annular outer retinopathy (AAOR). CONCLUSIONS The study was performed at a Polish referral center and may to some extent reflect the varied geographical distribution of white dot syndromes, as none of the subjects was found to suffer from acute zonal occult outer retinopathy (AZOOR), acute macular neuroretinopathy (AMN), or diffuse unilateral subacute neuroretinitis (DUSN). Long-term follow-up is warranted by the evolution of lesions in the eye fundus, while management depends on correct diagnosis of WDS. When the posterior pole is involved in some cases of the WDS an immunosuppressive treatment, the use of the PDT or anti-VEGF injections were necessary.


The role of cyclin E in cell proliferation, development and cancer.
Normal cell proliferation is under strict regulation governed by checkpoints located at distinct points in the cell cycle. The deregulation of these checkpoint events and the molecules associated with them may transform a normal cell into a cancer cell. One of these checkpoints whose deregulation results in transformation occurs at the Restriction point, near the G1/S boundary. The periodic appearance of one of the recently identified regulatory cyclins, cyclin E, coincides precisely with the timing of the Restriction point. The deregulation in the expression and activity of cyclin E has been associated with a number of cancers and is thought to be involved in the process of oncogenesis. In this chapter, we summarise the current knowledge on the regulation and apparent function of cyclin E in normal proliferating cells and in developing tissue and alterations of these processes in cancer.


Vibrationally averaged isotropic dispersion energy coefficients of the parahydrogen dimer.
We compare the sum-over-states and coupled cluster linear response formalisms for the determination of imaginary-frequency polarizabilities of H(2). Using both approaches, we compute isotropic dispersion energy coefficients C(n) (n = 6, 8, 10) for H(2)-H(2) molecular pairs over a wide range of H(2) bond lengths. We present vibrationally averaged dispersion energy coefficients for H(2)-H(2), H(2)-D(2), and D(2)-D(2) molecular pairs and examine the coefficients' convergence with respect to basis set.


Evolutionary adaptation to juvenile malnutrition impacts adult metabolism and impairs adult fitness in Drosophila.
Juvenile undernutrition has lasting effects on adult metabolism of the affected individuals, but it is unclear how adult physiology is shaped over evolutionary time by natural selection driven by juvenile undernutrition. We combined RNAseq, targeted metabolomics, and genomics to study the consequences of evolution under juvenile undernutrition for metabolism of reproductively active adult females of Drosophila melanogaster. Compared to Control populations maintained on standard diet, Selected populations maintained for over 230 generations on a nutrient-poor larval diet evolved major changes in adult gene expression and metabolite abundance, in particular affecting amino acid and purine metabolism. The evolved differences in adult gene expression and metabolite abundance between Selected and Control populations were positively correlated with the corresponding differences previously reported for Selected versus Control larvae. This implies that genetic variants affect both stages similarly. Even when well fed, the metabolic profile of Selected flies resembled that of flies subject to starvation. Finally, Selected flies had lower reproductive output than Controls even when both were raised under the conditions under which the Selected populations evolved. These results imply that evolutionary adaptation to juvenile undernutrition has large pleiotropic consequences for adult metabolism, and that they are costly rather than adaptive for adult fitness. Thus, juvenile and adult metabolism do not appear to evolve independently from each other even in a holometabolous species where the two life stages are separated by a complete metamorphosis.


Enhanced skin permeation of cationic drug ketotifen through excised guinea pig dorsal skin by surfactants with different electric charges.
Using excised guinea pig dorsal skin, we examined the effects of three surfactants, anionic sodium dodecylsulfate (SDS), cationic n-dodecyltrimethylammonium bromide and non-ionic n-dodecyl-beta-D-maltoside, all of which commonly have an n-dodecyl group, on in vitro skin permeation of the cationic drug ketotifen. All these three surfactants increased the skin permeation of ketotifen. Among the surfactants tested, anionic SDS had the largest enhancement effects, and significantly increased the permeation at concentrations over 1 mM. The enhancement effect of the same anionic surfactant on the permeation of anionic salicylate was smaller and similar to that of cationic n-dodecyltrimethylammonium. The enhancement effects of SDS on ketotifen permeation were more marked than those of the cationic surfactant but differed from previous findings of their effects on other drugs permeation. Analysis of the retention of ketotifen in the skin suggested that SDS-induced increase in the transfer of hydrophilic ketotifen to the skin is the main reason for the marked increase in skin permeation.


Effect of implant design on the initial biomechanical stability of two self-tapping dental implants.
This study evaluated the effect of two self-tapping implants on implant stability in immediate implantation. Two types of self-tapping implants, straight flute (STF) and spiral flute (SPF) designs, were studied. Two synthetic bone blocks with varying densities (0.32 g/cm3 and 0.16 g/cm3) were chosen to simulate the bone quality of the anterior maxilla. Insertion torque values were measured by a torque testing machine during implant insertion. Four biomechanical tests were performed: resonance frequency analysis was conducted using the Osstell device, and the strengths of screw push-in, lateral bending, and pull-out were evaluated using an MTS machine. The strength for each design feature was obtained by averaging the results of 10 trials. In total, 40 specimens were tested for each bone density. Statistical difference was determined by one-way analysis of variance followed by Bonferroni post hoc multiple tests between groups. The STF and SPF groups exhibited similar insertion torque values (p = 0.525 in low-density bone, and p = 0.99 in high-density bone). A significant difference (p < 0.001) was observed in the push-in test between the two groups when low-density bone was tested. The SPF group exhibited a significantly higher lateral bending force (p = 0.001) and a higher stiffness (p < 0.001) than the STF group in high-density bone. The SPF design attained higher (p < 0.001) ISQ numbers than the STF design, but all numbers were below 60. Implant stability can be influenced by the apical fixture design of self-tapping implants in immediate implantation.


[Bronchodilators--effective also in negative broncholysis test. Decreased dyspnea decreased in any case].
According to GOLD (Global Initiative for Chronic Obstructive Lung Disease), bronchodilator drugs are indicated for all degrees of severity of COPD. In stage 1, short-acting substances are administered as required; in stage 2, permanent treatment with short- or long-acting bronchodilators is applied. Patients who fail to respond to broncholysis testing can also benefit from such treatment over the long term. Combination treatment with beta 2 sympathomimetics and anticholinergic drugs makes good sense, since additive effects can develop. Since COPD patients often have CAD in addition, consideration must be given to cardiac side effects--in particular in the case of theophylline. The sole life-prolonging treatment for COPD continues to be the long-term administration of oxygen; the only measure capable of stopping progressive deterioration of lung function is cessation of smoking.


Carbon-Bonding Metal Catalysis (CBMC): A Supramolecular Complex Directs Structural-Isomer Selection in Gold-Catalyzed Reactions.
Carbon is a primary element to constitute organic molecules, while metal catalysis is a basic tool in organic synthesis. The establishment of a link between the ubiquitous carbon bonding and metal catalysis is thus a fundamentally important problem. However, there is yet no experimental example to introduce the role of carbon bonding in a metal catalysis process. Herein, we merged the topics of carbon bonding and metal catalysis together and demonstrated that a supramolecular carbon-bonding metal complex can not only give rise to catalytic activity but, more remarkably, direct structural-isomer selection events in gold-catalyzed reactions. The experimental results unveil the fact that the imposing of weak carbon-bonding interactions on a gold complex can alter the carbene as well as the Lewis acid property of these catalysts. These results illustrate a non-negligible role of weak carbon-bonding interactions in the modulation of metal catalysis. As such, carbon-bonding metal catalysis is suggested to be used as a routine tool not only in the development of reactions but more frequently in analyzing reaction processes in metal catalysis.


Alterations of visual contrast sensitivity in Parkinson's disease.
Contrast sensitivity functions were determined in a population of 18 patients suffering from Parkinson's disease, and compared with the data obtained in an age-matched group of healthy controls. The controls were more sensitive at all spatial frequencies tested than the patients. The statistical comparisons were highly significant, indicating general differences between the PD patients and the controls not related to individual spatial frequency channels. When comparing the sensitivity loss between low and high spatial frequencies no significant differences were found suggesting that the decrease in contrast sensitivity is a global effect. We controlled for effects of age and cerebral atrophy, and our findings cannot be accounted for by these factors. In addition, the amount of contrast sensitivity loss was not correlated with the severity of the disease. These global functional alterations appear to be related to the reduction of dopamine at various sites of the visual system.


[Effects of inpatient preventive and rehabilitative measures for mothers and their children - a controlled comparison trial].
Rehabilitation measures have for a long time been evaluated primarily by means of pre-post comparisons of the group examined or by comparing different intervention programmes. This study was aimed at examining the current status and medium-term changes of mothers' health and psychosocial situation, comparing mothers with or without approval and participation in an inpatient mother-child rehabilitation measure. The sample consisted of 477 women. The first survey (T1) took place at the time of application; the postal follow-up survey (T2) was carried out 6 months later. At this point 353 mothers had completed their measure (IG=intervention group); 75 women had not yet started (WG=waiting group), and 49 applications had finally been rejected (AG=rejected group). This study examines how far the 3 groups differ with respect to their health parameters at the time of application as well as 6 months later. The outcome variables, containing health complaints, somatic and mental illnesses, psychosocial factors (e. g., single parenthood), or restrictions of daily activities and the range of participation in family and job affairs, were selected following the assessment guidelines of the Medical Review Board of the Statutory Health Insurance Funds (MDS). Duration and severity of symptoms were measured with Likert scales (indirect measurement of change), the mental health status with SCL-K-9. At the time of application the 3 groups of mothers did not differ significantly with respect to their complaints and symptoms, but some minor tendencies became apparent. Mothers, who were admitted for the intervention (IG and WG) showed more complaints and symptoms than mothers, whose applications had been rejected (AG). 6 months later, highly significant changes and clinically important improvements were observed for IG mothers: large effects were achieved regarding the number of illnesses, health complaints, the degree of daily activities and participation as well as mental health of participants. Minor effects were obtained with respect to psychosocial strains. In contrast the number of illnesses and health complaints among mothers, who had not (yet) participated, increased within half a year. Similarly, psychological strains were greater than before: in the waiting group they had risen significantly, in the rejected group values resembled those of the intervention group before intervention. Inpatient measures do not only cause short-term improvements of mothers' health status, they also lead to clinically relevant reductions of health impairments and complaints in the medium term. In contrast, the health status of mothers, who miss the intervention, deteriorates. This also applies to mothers, who were rejected. Therefore, it is highly recommended to take the self-reporting forms into account when identifying mothers for the intervention, and also to start the intervention early after the approval.


Male Nurses in Israel: Barriers, Motivation, and How They Are Perceived by Nursing Students.
The current worldwide nursing shortage remains a challenge for the nursing profession. Encouraging men to become nurses and, thereby, increasing the number of practitioners are crucial factors in facing this challenge. The historiography of nursing presents nursing as "women's work," based on the assumption that it is inherently appropriate for women only. Although men were employed as nurses even before nursing was recognized as a profession, male nurses were always a minority in the field. Over the years, the proportion of male nurses has increased, but they still comprise only 5 to 10% of the nursing workforce in the western world. This study examined men's motives for a career choice of nursing, how male nurses are perceived, and the barriers that they face. The study was conducted among 336 nursing students studying in a co-educational program in various academic tracks at a public, nonsectarian university in the south of Israel. Participants completed the following questionnaires in one study session: sociodemographic questionnaire; Attitudes Towards Men in Nursing Scale; motives for career choice questionnaire; and the questionnaire of the perceptions of the professional status of nursing. Study findings revealed that men tended to choose nursing because of financial constraints significantly more frequently than women (P=.001). Among the participants, there was no significant between-sex difference in the perception nursing as women's work (P=.002) or in perception of male nurses as homosexuals. Results of the study showed that the status of the nursing profession is considered low, and the low status deters men from choosing nursing as a career. The motivation for men's career choice must be understood, and men must be empowered to improve their work conditions and financial remuneration in order to recruit men to the field and to improve the perception of the profession and its public status.


[Chemotherapy for CRPC].
Cabazitaxel, new chemotherapeutic agent for castration resistant prostate cancer (CRPC) treated after docetaxel, was developed. In addition, new hormonal drugs for CRPC, such as enzalutamide and abiraterone were also approved in Japan recently. Treatment strategy for CRPC using these drugs is still controversial, therefore we need much more clinical data of Japanese patients with CRPC. Management of this severe condition and future of prostate cancer were discussed.


Rh-catalysed [5 + 1] cycloaddition of allenylcyclopropanes and CO: reaction development and application to the formal synthesis of (-)-galanthamine.
A Rh-catalysed [5 + 1] cycloaddition of allenylcyclopropanes and CO has been developed to synthesize functionalized 2-methylidene-3,4-cyclohexenones. The scope of this methodology has been investigated, showing that various functional groups can be tolerated. Both di- and tri-substituted allenylcyclopropanes can be applied to this cycloaddition and the [5 + 1] cycloadducts with the E configuration were obtained as the major products. In addition, the present [5 + 1] cycloaddition reaction has been utilized as a key step in the formal synthesis of the natural product (-)-galanthamine.


Heparan Sulfate Proteoglycan Is an Important Attachment Factor for Cell Entry of Akabane and Schmallenberg Viruses.
Akabane virus (AKAV) and Schmallenberg virus (SBV) are members of the genus Orthobunyavirus, which are transmitted by arthropod vectors with a broad cellular tropism in vitro as well as in vivo Both AKAV and SBV cause arthrogryposis-hydranencephaly syndrome in ruminants. The main cellular receptor and attachment factor for entry of these orthobunyaviruses are unknown. Here, we found that AKAV and SBV infections were inhibited by the addition of heparin or enzymatic removal of cell surface heparan sulfates. To confirm this finding, we prepared heparan sulfate proteoglycan (HSPG)-knockout (KO) cells by using a clustered regularly interspaced short palindromic repeat (CRISPR)-Cas9 system and measured the quantities of binding of these viruses to cell surfaces. We observed a substantial reduction in AKAV and SBV binding to cells, limiting the infections by these viruses. These data demonstrate that HSPGs are important cellular attachment factors for AKAV and SBV, at least in vitro, to promote virus replication in susceptible cells.IMPORTANCE AKAV and SBV are the etiological agents of arthrogryposis-hydranencephaly syndrome in ruminants, which causes considerable economic losses in the livestock industry. Here, we identified heparan sulfate proteoglycan as a major cellular attachment factor for the entry of AKAV and SBV. Moreover, we found that heparin is a strong inhibitor of AKAV and SBV infections. Revealing the molecular mechanisms of virus-host interactions is critical in order to understand virus biology and develop novel live attenuated vaccines.


Structure and Dynamics of the Rhomboid Protease GlpG in Liposomes Studied by Solid-State NMR.
Rhomboid proteases are intramembrane proteases that hydrolyze substrate peptide bonds within the lipid bilayer and are important for a wide range of biological processes. The bacterial intramembrane protease GlpG is one of the model systems for structural investigations of the rhomboid family. Two different models of substrate gating have been proposed, based on crystal structures of GlpG in detergent micelles. Here, we present a detailed investigation of enzymatically active GlpG in a native-like lipid environment using solid-state NMR spectroscopy. Proton-detected experiments confirm the presence of water molecules in the catalytic cavity. A secondary chemical shift analysis indicates a previously unobserved kink in the central part of the gating helix TM5. Dynamics measurements revealed a dynamic hotspot of GlpG at the N-terminal part of TM5 and the adjacent loop L4, indicating that this region is important for gating. In addition, relaxation dispersion experiments suggest that TM5 is in conformational exchange between an open and a closed conformation.


Longitudinal Metagenomic Analysis of Hospital Air Identifies Clinically Relevant Microbes.
We describe the sampling of sixty-three uncultured hospital air samples collected over a six-month period and analysis using shotgun metagenomic sequencing. Our primary goals were to determine the longitudinal metagenomic variability of this environment, identify and characterize genomes of potential pathogens and determine whether they are atypical to the hospital airborne metagenome. Air samples were collected from eight locations which included patient wards, the main lobby and outside. The resulting DNA libraries produced 972 million sequences representing 51 gigabases. Hierarchical clustering of samples by the most abundant 50 microbial orders generated three major nodes which primarily clustered by type of location. Because the indoor locations were longitudinally consistent, episodic relative increases in microbial genomic signatures related to the opportunistic pathogens Aspergillus, Penicillium and Stenotrophomonas were identified as outliers at specific locations. Further analysis of microbial reads specific for Stenotrophomonas maltophilia indicated homology to a sequenced multi-drug resistant clinical strain and we observed broad sequence coverage of resistance genes. We demonstrate that a shotgun metagenomic sequencing approach can be used to characterize the resistance determinants of pathogen genomes that are uncharacteristic for an otherwise consistent hospital air microbial metagenomic profile.


Level of surgical experience is associated with change in hip center of rotation following cementless total hip arthroplasty: A radiographic assessment.
After total hip arthroplasty (THA), restoration of hip center of rotation (COR) is essential to ensure stability of the prosthetic hip and longevity of the prosthesis. Our aim was to determine whether, and how, the COR changed postoperatively compared to the native COR following implantation of a cementless acetabular component in anatomical position and to compare the accuracy of cup placement between two surgeons with different levels of surgical experience. We evaluated 145 patients (145 hips) who underwent unilateral primary THA, who had no distorted acetabulum on the affected hip and a normal contralateral hip. Hip reconstruction was radiologically and clinically assessed at a minimum 2-year follow-up. Perioperative change in COR, initial cup position, offset, leg-length discrepancy (LLD), radiographic cup orientation, Harris Hip Score (HHS), component loosening, and dislocations were compared between the highly experienced surgeon and less-experienced surgeon groups. The COR was significantly displaced in the superior and medial directions postoperatively. Significant differences were identified in the vertical COR change, initial cup position, LLD, cup inclination, and cups within safe zones, but not in the horizontal COR change, offset parameters, cup anteversion, or HHS. There were no radiographic evidence of component loosening in both groups, but three dislocations (7%) only in the group operated on by the less-experienced surgeon (p = 0.027). We found that the postoperative COR tended to be displaced in the superior and medial directions, and that the level of surgical experience strongly affected the accuracy and consistency of cup placement, particularly in COR position and cup inclination.


Clinical Study on the Treatment of Complex Anal Fistula by Phased Chinese Herbal Sitz Bath Based on "Fuzheng Quxie" Theory.
To evaluate the clinical efficacy of a phased Chinese herbal sitz bath for the treatment of complex anal fistula based on the traditional Chinese medicine theory of "Fuzheng Quxie." We enrolled 200 patients with complex anal fistula who were hospitalized and treated surgically in the Department of Anorectal Medicine of Suining Central Hospital from June 2021 to December 2022. The patients were randomly divided into 2 groups of 100 patients each: sitz bath group 1 (postoperation, the patients used a sitz bath with a self-formulated decoction of "Qingre Zaoshi Zhitong" formula, once in the morning and once in the evening) and sitz bath group 2 (postoperation, the patients used the same decoction as the sitz bath group 1, and when the incision began to heal, the patients then used a sitz bath with a self-formulated decoction of "Yiqi Fuzheng" formula, once in the morning and once in the evening). Both groups were treated for 6 weeks. The assessments included wound symptom score, incision healing time, clinical efficacy after treatment, and serum-related growth factor concentrations and anorectal dynamics indicators before and after treatment. After treatment, sitz bath group 2 had less wound pain, edema, exudate, and granulation growth scores than sitz bath group 1 (P < .05). Postoperative incision healing time was faster in sitz bath group 2 22.96 (1.96) d than in sitz bath group 1 26.19 (2.62) d (t = 9.872, P < .001). The total effective rate after treatment was higher in sitz bath group 2 96% (96/100) than in sitz bath group 1 88% (88/100) (χ2 = 4.4, P = .04). Serum transforming growth factor β, epidermal growth factor, and basic fibroblast growth factor concentrations were higher in both groups after treatment than before treatment, and the concentrations of the growth factors after treatment were higher for sitz bath group 2 than for sitz bath group 1 (P < .001). Anal rest pressure and anal maximum contraction pressure were higher in both groups after treatment than before treatment, and the pressures after treatment were higher for sitz bath group 2 than for sitz bath group 1 (P < .05). Based on the theory of Fuzheng Quxie, the effectiveness and superiority of a phased Chinese herbal sitz bath for the treatment of complex anal fistula were remarkable, which could promote the improvement of wound symptoms and anal function. This may be related to increases in growth factor concentrations and anorectal dynamics indicators.


Effect of contiguity and figure-ground organization on the area rule of lightness.
In a simple two-dimensional (2D) display composed of two uniform surfaces with different luminances, the lightness of the darker surface varies as a function of its relative area while its luminance is held constant (Gilchrist & Radonjić, 2009; Li & Gilchrist, 1999). This phenomenon is known as the area rule of lightness, and although it is extensively studied in the literature, the underlying principles are still largely unknown. Here, using computer-generated stimuli, we investigated the effects of contiguity and figure-ground organization on the area rule of lightness. Stimuli were 2D disks composed of radial sectors with high (25 cd/m2) or low (8 cd/m2) luminance. On each trial, observers judged the lightness of the sectors by adjusting the luminance of a matching patch. Four conditions were tested. In the contiguous condition, there were one dark and one light sector; in the noncontiguous condition, both the light and dark surfaces were split into four equal radial sectors. Figure and ground conditions were generated by adding small contextual elements to the stimulus. We found that the area rule applied under all conditions; however, the functional form of the effect showed marked differences across conditions. Taken together, our results show that both high-level (e.g., perceptual grouping, figure-ground organization) and low-level (e.g., spatial-summation) mechanisms play a role in the area rule of lightness.


Angiotensin II responses in AT1A receptor-deficient mice: a role for AT1B receptors in blood pressure regulation.
Most of the classic functions of the renin-angiotensin system are mediated by type 1 (AT1) angiotensin receptors, of which two subtypes, AT1A and AT1B, have been identified. However, distinct functions for these two AT1 receptors have been difficult to separate. We examined the pressor effects of angiotensin II in Agtr1A -/- mice, which lack AT1A receptors. In enalapril-pretreated Agtr1A -/- mice, angiotensin II caused significant and dose-proportional increases in mean arterial pressure. This pressor response was not blocked by pretreatment with sympatholytic agents but was completely inhibited by the AT1-receptor antagonists, losartan and candesartan, suggesting that it is directly mediated by AT1B receptors. Chronic treatment of Agtr1A -/- mice with losartan reduced systolic blood pressure from 80 +/- 5 to 72 +/- 4 mmHg (P < 0.04), suggesting a role for AT1B receptors in chronic blood pressure regulation. These studies provide the first demonstration of in vivo pressor effects mediated by AT1B receptors and demonstrate that, when AT1A receptors are absent, the AT1B receptor contributes to the regulation of resting blood pressure.


Physical activity and health-related quality of life in multiple myeloma survivors: the PROFILES registry.
To investigate whether physical activity (PA) is associated with health-related quality of life (HRQOL) outcomes in multiple myeloma (MM) survivors up to 11 years after diagnosis. We used data from the Patient Reported Outcomes Following Initial treatment and Long term Evaluation of Survivorship registry. We included 175 MM survivors diagnosed between 1999 and 2009 as registered by the Netherlands Cancer Registry. Sixty-four per cent (n=112/175) of patients who received the questionnaires, completed the European Organisation for Research and Treatment of Cancer (EORTC) QLQ-C30 and the EORTC QLQ-MY20. Patients were classified into two groups: physically active and not physically active patients. Univariable and multivariable linear regression models were used to evaluate associations between PA and HRQOL outcomes. Physically active patients reported a statistically significant higher global health status/HRQOL (p=0.001), lower fatigue (p=0.002) and fewer side effects of treatments (p=0.001), than not physically active patients. PA was not associated with psychological symptoms (ie, anxiety and depressive symptoms) (anxiety: p=0.139; depressive symptoms: p=0.073). Exploratory analyses performed on the other scales of the EORTC QLQ-C30 indicated statistically significant better outcomes in several functional and symptom subscales for physically active patients. These findings might contribute to a better understanding of the relationship between PA and disease specific HRQOL aspects in MM survivors. Prospective studies are warranted to further elucidate on the beneficial effects of PA on HRQOL outcomes of MM survivors.


Stability of φ^{4}-vector model: Four-loop ε expansion study.
In most papers, φ^{4}-field theory with the vector (d-component) field φ_{α} is considered as a particular case of the n-component field model for n=d and O(n) symmetry. However, in such a model the symmetry O(d) admits an addition to the action of a term proportional to the squared divergence of the field ∼h(∂_{α}φ_{α})^{2}. From the point of view of renormalization group analysis, it requires a separate consideration, because it may well change the nature of the critical behavior of the system. Therefore, this frequently neglected term in the action requires a detailed and accurate study on the issue of the existence of new fixed points and their stability. It is known that within the lower order of perturbation theory the only infrared stable fixed point with h=0 exists but the corresponding positive value of stability exponent ω_{h} is tiny. This led us to analyze this constant in higher orders of perturbation theory by calculating the four-loop renormalization group contributions for ω_{h} in d=4-2ɛ within the minimal subtraction scheme, which should be enough to infer positivity or negativity of this exponent. The value turned out to be undoubtedly positive, although still small even in higher loops: 0.0156(3). These results cause the corresponding term to be neglected in the action when analyzing the critical behavior of the O(n)-symmetric model. At the same time, the small value of ω_{h} shows that the corresponding corrections to the critical scaling are significant in a wide range.


Paramecium aurelia species complex in the catchment area of the River Raba mountain course (the Carpathians).
In the water bodies of the Beskid Wyspowy Mts three species of the Paramecium aurelia complex have been found, i.e. P. biaurelia, P. tetraurelia and P. novaurelia. P. tetraurelia, which in Europe is a very rare species, was for the first time recorded in this region of the Carpathians.


Comparative study of 5% permethrin cream and 1% lindane lotion for the treatment of scabies.
A multicenter, randomized, investigator-blind controlled trial was conducted to compare the safety and efficacy of a single, whole-body application of 5% permethrin cream with that of 1% lindane lotion for the treatment of scabies in 467 patients. At 14 +/- 3 days after treatment, the mean active lesion count decreased from pretreatment levels of 85 (range, 4 to 600) in both treatment groups to 14 (range, 0 to 133) in the permethrin group and to 15 lesions (range, 0 to 500) in the lindane group. At 28 +/- 7 days after treatment, complete resolution had occurred in 181 (91%) of 199 patients treated with permethrin and in 176 (86%) of 205 patients given lindane. Pruritus due to scabies persisted at 28 +/- 7 days in 14% of the permethrin group and in 25% of the lindane group. The most frequent adverse effects were new or increased pruritus and mild, transient burning or stinging; the latter was slightly more frequent following permethrin treatment and appeared to be related to severity of infestation. Because of a lower potential for neurologic toxicity, permethrin may be preferable to lindane for the treatment of scabies particularly in children.


EVI/WLS function is regulated by ubiquitylation and is linked to ER-associated degradation by ERLIN2.
WNT signalling is important for development in all metazoans and is associated with various human diseases. The ubiquitin-proteasome system (UPS) and regulatory endoplasmic reticulum-associated degradation (ERAD) have been implicated in the production of WNT proteins. Here, we investigated how the WNT secretory factor EVI (also known as WLS) is ubiquitylated, recognised by ERAD components and subsequently removed from the secretory pathway. We performed a focused immunoblot-based RNAi screen for factors that influence EVI/WLS protein stability. We identified the VCP-binding proteins FAF2 and UBXN4 as novel interaction partners of EVI/WLS and showed that ERLIN2 links EVI/WLS to the ubiquitylation machinery. Interestingly, we also found that EVI/WLS is ubiquitylated and degraded in cells irrespective of their level of WNT production. This K11, K48 and K63-linked ubiquitylation is mediated by the E2 ubiquitin-conjugating enzymes UBE2J2, UBE2K and UBE2N, but is independent of the E3 ubiquitin ligases HRD1 (also known as SYVN1) and GP78 (also known as AMFR). Taken together, our study identifies factors that link the UPS to the WNT secretory pathway and provides mechanistic details of the fate of an endogenous substrate of regulatory ERAD in mammalian cells. This article has an associated First Person interview with the first author of the paper.


Modulation of osteoblast function using poly(D,L-lactic acid) surfaces modified with alkylation derivative of chitosan.
Poly(D,L-lactic acid) (PDLLA) was modified with alkylated chitosan (N-butyl chitosan and N-cetyl chitosan), and the effects of modified films on the functions of rat osteoblasts were investigated. The characteristics of surfaces (both modified and control) were examined by water contact angle measurement and electron spectroscopy for chemical analysis (ESCA). Cell morphologies on these surfaces were taken using scanning electron microscopy (SEM). Cell attachment and proliferation were used to assess cell behavior on modified surface and control. MTT assay was used to determined cell viability, and alkaline phosphatase (ALP) activity was taken to evaluate differentiated cell function. Compared with the untreated films, no significant difference in cell attachment of osteoblasts was found on the modified films at a period of 8 h (p > 0.05). However, cell proliferation of N-butyl chitosan rather than N-cetyl chitosan modified PDLLA films was significantly higher than that found on control one (p < 0.05) at the end of the 4th and 7th days. The cell viability of osteoblasts on N-butyl chitosan modified PDLLA films were found higher than that on control (p < 0.05). These results suggested that N-butyl chitosan contributed greater than N-cetyl chitosan when used to modify PDLLA films for improving its biocompatibility.


Stress-induced hyperthermia and basal body temperature are mediated by different 5-HT(1A) receptor populations: a study in SERT knockout rats.
Disturbances in the serotonergic system are implicated in many central nervous system disorders. The serotonin transporter (SERT) regulates the serotonin homeostasis in the synapse. We recently developed a rat which lacks the serotonin transporter (SERT(-/-)). It is likely that adaptive changes take place at the level of pre- and postsynaptic 5-HT receptors. Because autonomic responses are often used to measure 5-HT(1A) receptor function, we analysed these responses by examining the effects of a 5-HT(1A) receptor agonist and antagonist under in vivo conditions in the SERT(-/-) rat. Moreover, we studied the effect of a mild stressor on the body temperature (stress-induced hyperthermia) because of the known involvement of 5-HT(1A) receptors in this phenomenon. Results show that core body temperature did not differ between genotypes under basal, non-stressed conditions. Compared to SERT(+/+) rats, stress-induced hyperthermia was reduced in SERT(-/-) rats. The 5-HT(1A) receptor agonist [R(+)-N-(2[4-(2,3-dihydro-2-2-hydroxy-methyl-1,4-benzodioxin-5-yl)-1-piperazininyl]ethyl)-4-fluorobenzoamide HCl (flesinoxan) reduced stress-induced hyperthermia in both genotypes. The flesinoxan-induced hypothermia in SERT(+/+) rats was blocked by the 5-HT(1A) receptor antagonist [N-(2-[4-(2-methoxyphenyl)-1-piperazinyl]ethyl)-N-(2-pyridinyl) cyclohexane carboxamide 3HCl (WAY100635). Moreover, WAY100635-induced hyperthermia in SERT(-/-), but not in SERT(+/+) rats. In SERT(-/-) rats, WAY100635 completely blocked the flesinoxan-induced reduction of stress-induced hyperthermia. Interestingly, flesinoxan-induced hypothermia was absent in SERT(-/-) rats. It is concluded that the SERT knockout rat reveals that 5-HT(1A) receptors modulating stress-induced hyperthermia belong to a population of receptors that differs from that involved in hypothermia.


Collinearity between a 30-centimorgan segment of Arabidopsis thaliana chromosome 4 and duplicated regions within the Brassica napus genome.
Arabidopsis thaliana (the model dicotyledonous plant) is closely related to Brassica crop species. Genome collinearity, or conservation of marker order, between Brassica napus (oilseed rape) and A. thaliana was assessed over a 7.5-Mbp region of the long arm of A. thaliana chromosome 4, equivalent to 30 cM. Estimates of copy number indicated that sequences present in a single copy in the haploid genome of A. thaliana (n = 5) were present in 2-8 copies in the haploid genome of B. napus (n = 19), while sequences present in multiple copies in A. thaliana were present in over 10 copies in B. napus. Genetic mapping in B. napus of DNA markers derived from a segment of A. thaliana chromosome 4 revealed duplicated homologous segments in the B. napus genome. Physical mapping in A. thaliana of homologues of Brassica clones derived from these regions confirmed the identity of six duplicated segments with substantial homology to the 7.5-Mbp region of chromosome 4 in A. thaliana. These six duplicated Brassica regions (on average 22 cM in length) are collinear, except that two of the six copies contain the same large internal inversion. These results have encouraging implications for the feasibility of shuttling between the physical map of A. thaliana and genetic maps of Brassica species, for identifying candidate genes and for map based gene cloning in Brassica crops.


Referenced Kendrick Mass Defect Annotation and Class-Based Filtering of Imaging MS Lipidomics Experiments.
Because of their diverse functionalities in cells, lipids are of primary importance when characterizing molecular profiles of physiological and disease states. Imaging mass spectrometry (IMS) provides the spatial distributions of lipid populations in tissues. Referenced Kendrick mass defect (RKMD) analysis is an effective mass spectrometry (MS) data analysis tool for classification and annotation of lipids. Herein, we extend the capabilities of RKMD analysis and demonstrate an integrated method for lipid annotation and chemical structure-based filtering for IMS datasets. Annotation of lipid features with lipid molecular class, radyl carbon chain length, and degree of unsaturation allows image reconstruction and visualization based on each structural characteristic. We show a proof-of-concept application of the method to a computationally generated IMS dataset and validate that the RKMD method is highly specific for lipid components in the presence of confounding background ions. Moreover, we demonstrate an application of the RKMD-based annotation and filtering to matrix-assisted laser desorption/ionization (MALDI) IMS lipidomic data from human kidney tissue analysis.


HIV and pneumococcal disease.
To describe the impact of highly active antiretroviral therapy on the burden of pneumococcal disease and advances in our understanding of the impact of HIV on this disease. Although highly active antiretroviral therapy has reduced the burden of pneumococcal disease among HIV-infected adults, these infections remain far more common than in HIV uninfected adults. HIV-infected adults who smoke or have comorbidities are at particular risk. In the absence of highly active antiretroviral therapy, pneumococcal meningitis has emerged in Africa as a major disease burden with a high mortality among HIV-infected children and adults. Conjugate pneumococcal vaccine protects HIV-infected infants from pneumococcal pneumonia. In the United States, where conjugate vaccine is given to children, herd immunity has reduced the burden of invasive pneumococcal disease among HIV-infected adults. The pneumococcus remains a significant cause of morbidity and mortality among HIV-infected children and adults, both in developed and in developing countries.


[Pulmonary to systemic flow ratios in patients with ventricular septal defect: estimation by transmitral flow velocity].
Using pulsed Doppler echocardiography, left ventricular inflow flow volume (LVIV) and outflow flow volume (LVOV) were noninvasively determined, and the ratio of pulmonary to systemic flow (Qp/Qs) was evaluated as the ratio of LVIV to LVOV (LVIV/LVOV). Thirty patients with ventricular septal defect (VSD) were studied, and 47 cardiac patients without aortic or mitral valve disease or intracardiac shunt served as controls. LVOV was derived from the left ventricular ejection flow velocity and the outflow tract diameter immediately proximal to the aortic valve ring. LVIV was derived from the transmitral flow velocity and the M-mode tracing of mitral valve motion. Doppler-determined cardiac outputs (COin and COout) were calculated as the products of LVIV or LVOV as and heart rates. Cardiac outputs were also determined by the dye dilution method (COdye) references for comparison with Doppler-determined cardiac outputs. There were good correlations between COdye and COin (y = 1.18x-243, r = 0.85, p less than 0.005, SEE = 1026 ml/min) and between COdye and COout (y = 1.16x-323, r = 0.90, p less than 0.005, SEE = 639 ml/min). LVIV and LVOV correlated well in the controls (y = 0.95x + 5.3, r = 0.94, p less than 0.005, SEE = 6.6 ml). LVIV/LVOV was 0.97 +/- 0.1 (mean +/- SD) in the controls; whereas LVIV/LVOV (1.86 +/- 0.90) was significantly higher in patients with VSD (p less than 0.01) and this ratio correlated well with Qp/Qs by an oximetry (r = 0.98, SEE = 0.20, n = 14), including patients associated with pulmonary regurgitation. These findings indicate that our method permits determination of LVIV with a high degree of accuracy and that the Doppler-determined LVIV/LVOV is clinically useful to evaluate accurately the magnitude of shunt flows in patients with VSD.


In vitro teratology.
The purpose of this conference was to reevaluate the need for and use of in vitro teratology assays; to examine the validation process for in vitro tests; and to discuss progress in the validation of in vitro teratology screens. Participants enthusiastically supported further development of short-term in vivo and in vitro systems both as prescreens for developmental toxicity and as experimental systems to explore mechanisms of action of toxicants. The group strongly endorsed the development of an updated reference list ("gold standard") of known developmental toxicants and nontoxicants as essential to further progress in developing and validating prescreening efforts. Independently, an expert group should further evaluate the performance characteristics for a validated prescreen. The limits of usefulness of prescreens for product development, regulatory use, and mechanistic investigations need to be clearly defined. Finally, too few in vitro teratology prescreens have been evaluated under multiple-laboratory conditions with common, agreed-upon test agents to draw firm conclusions regarding the merit and reproducibility of in vitro teratology prescreens. There was general agreement regarding the need to move several of the assays further along the validation pathway, at least using a short list of reference compounds.


Effects of childhood chronic illness on families.
This study examined the intellectual ability, academic performance, behavior, and social perceptions of children with a seizure disorder and their nonafflicted siblings. These data suggest the children are at risk for developmental lag and related problems. Impact of the illness on the family differentiated the sibling pair on many tasks and on parents' feelings towards their children. Although parents reported providing a normal environment for their children, the children reported their perceptions of the home environment to be further from the (population norm) than their parents. These results provide evidence for the whole family being affected by the chronic illness thus supporting social work's inherent holistic view. It also underscores the need for a secondary preventive approach with all families containing a child with a handicapping condition.


Visual Feedback Modulates Aftereffects and Electrophysiological Markers of Prism Adaptation.
Prism adaptation (PA) is both a model for visuomotor learning and a promising treatment for visuospatial neglect after stroke. The task involves reaching for targets while prism glasses horizontally displace the visual field. Adaptation is hypothesized to occur through two processes: strategic recalibration, a rapid self-correction of pointing errors; and spatial realignment, a more gradual adjustment of visuomotor reference frames that produce prism aftereffects (i.e., reaching errors upon glasses removal in the direction opposite to the visual shift). While aftereffects can ameliorate neglect, not all patients respond to PA, and the neural mechanisms underlying successful adaptation are unclear. We investigated the feedback-related negativity (FRN) and the P300 event-related potential (ERP) components as candidate markers of strategic recalibration and spatial realignment, respectively. Healthy young adults wore prism glasses and performed memory-guided reaching toward vertical-line targets. ERPs were recorded in response to three different between-subject error feedback conditions at screen-touch: view of hand and target (Experiment 1), view of hand only (Experiment 2), or view of lines to mark target and hand position (view of hand occluded; Experiment 3). Conditions involving a direct view of the hand-produced stronger aftereffects than indirect hand feedback, and also evoked a P300 that decreased in amplitude as adaptation proceeded. Conversely, the FRN was only seen in conditions involving target feedback, even when aftereffects were smaller. Since conditions producing stronger aftereffects were associated with a phase-sensitive P300, this component may index a "context-updating" realignment process critical for strong aftereffects, whereas the FRN may reflect an error monitoring process related to strategic recalibration.


Calibration-free absolute frequency response measurement of directly modulated lasers based on additional modulation.
A calibration-free electrical method is proposed for measuring the absolute frequency response of directly modulated semiconductor lasers based on additional modulation. The method achieves the electrical domain measurement of the modulation index of directly modulated lasers without the need for correcting the responsivity fluctuation in the photodetection. Moreover, it doubles measuring frequency range by setting a specific frequency relationship between the direct and additional modulation. Both the absolute and relative frequency response of semiconductor lasers are experimentally measured from the electrical spectrum of the twice-modulated optical signal, and the measured results are compared to those obtained with conventional methods to check the consistency. The proposed method provides calibration-free and accurate measurement for high-speed semiconductor lasers with high-resolution electrical spectrum analysis.


C5-Alkyl-2-methylurea-Substituted Pyridines as a New Class of Glucokinase Activators.
Glucokinase (GK) activators represent a class of type 2 diabetes therapeutics actively pursued due to the central role that GK plays in regulating glucose homeostasis. Herein we report a novel C5-alkyl-2-methylurea-substituted pyridine series of GK activators derived from our previously reported thiazolylamino pyridine series. Our efforts in optimizing potency, enzyme kinetic properties, and metabolic stability led to the identification of compound 26 (AM-9514). This analogue showed a favorable combination of in vitro potency, enzyme kinetic properties, acceptable pharmacokinetic profiles in preclinical species, and robust efficacy in a rodent PD model.


Effects of three month nasal continuous positive airway pressure treatment on electrocardiographic, echocardiographic and overnight polysomnographic parameters in newly diagnosed moderate/severe obstructive sleep apnea patients.
The objective of the study was to determine the effects of nasal continuous positive airway pressure (nCPAP) therapy on left ventricular (LV) function and electrocardiographic parameters in newly diagnosed moderate/severe obstructive sleep apnea (OSA) patients without cardiovascular comorbidities and medical treatments. We examined 44 patients who underwent overnight polysomnography together with 24-hour Holter electrocardiography, cardiopulmonary exercise testing including heart rate recovery at 1 minute (HRR-1), echocardiography, surface electrocardiography, and those who were diagnosed with moderate/severe OSA apnea--hypopnea index ≥ 15. After 3 months of nCPAP treatment, the above-mentioned examinations were repeated. Forty-four patients completed the treatment period. Twelve weeks on effective nCPAP induced a significant increase in the mitral E/A ratio (P = 0.001), as well as reductions in isovolumic relaxation time (P = 0.001) and mitral deceleration time (DT) (P = 0.002). There were no significant differences in LV ejection fraction, LV mass index, and pulsed wave Doppler parameters. Mean heart rate was 79.2 ± 12.5 pulses/minute, maximum P-wave duration 117.5 ± 8.6 msec, P-wave dispersion (PWd) 54.6 ± 10.2 msec, corrected QT interval (QTc) 436.5 ± 40.5 msec, and QT dispersion (QTd) 46.3 ± 7.1 msec, which significantly decreased to 70.4 ± 9.6 pulses/minute (P < 0.001), 111.5 ± 8.7 msec (P < 0.001), 51.6 ± 8.9 msec (P < 0.001), 418.4 ± 31.2 msec (P < 0.001), and 33.8 ± 3.4 msec (P < 0.001), respectively. Exercise capacity at baseline determined as 10.5 ± 2.2 metabolic equivalents (METS) and HRR-1 (20.6 ± 11.7 bpm) significantly increased (12.1 ± 1.5 METS and 27.4 ± 8.6 bpm). There was no significant difference in aortic root parameters. Three-month nCPAP therapy significantly increased LV shortening fraction, with no effect on systolic function or aortic root diameters and a positive effect on heart rate, PWd, HRR-1, QTc and QTd time following nCPAP therapy.


Japanese haemodialysis anaemia management practices and outcomes (1999-2006): results from the DOPPS.
Japanese haemodialysis (HD) patients not only have a very low mortality and hospitalization risk but also low haemoglobin (Hb) levels. Internationally, anaemia is associated with mortality, hospitalization and health-related quality of life (QoL) measures of HD patients. Longitudinal data collected from 1999 to 2006 from 60 to 64 representative Japanese dialysis units participating in the Dialysis Outcomes and Practice Patterns Study (DOPPS) were used to describe anaemia management practices and outcomes for Japanese HD patients. From 1999 to 2006, patient mean Hb increased from 9.7 g/dl to 10.4 g/dl, and the percentage of facilities with median Hb >or=10 g/dl increased from 27% to 75%. Hb was measured in the supine position for 90% of patients, resulting in substantially lower reported Hb values than those seen in other countries. As of 2006, erythropoietin (Epo) was prescribed to 83% of HD patients; mean Epo dose was 5231 units/week; intravenous (IV) iron use was 33% and median IV iron dose was 160 mg/month. Many patient- and facility-level factors were significantly related to higher Hb. A consistent overall pattern of lower mortality risk with higher baseline Hb levels was seen (RR = 0.89 per 1 g/dl higher Hb, P = 0.003). Facilities with median Hb >or=10.4 displayed a lower mortality risk (RR = 0.77, P = 0.03) versus facility median Hb <10.4 g/dl. Lower Hb levels were not significantly related to hospitalization risk, but were associated with lower QoL scores. These results provide detailed information on anaemia management practices in Japan and the relationships of anaemia control with outcomes, with implications of anaemia management worldwide.


The von Hippel-Lindau gene and protein in tumorigenesis and angiogenesis: a potential target for therapeutic designs.
The von Hippel-Lindau (VHL) protein is able to suppress tumor growth and to down-regulate many angiogenic factors, and is ubiquitously detected in adult and fetal tissues. This makes VHL an excellent target for therapeutic intervention. Observation of VHL alterations in sporadic tumors has been increasing as a result of examination of abnormalities other than intragenic mutations. These abnormalities include loss of chromosome 3p25, changes in the promoter, down-regulation of transcript, and changes in protein level. This article also presents the finding of differential expression of two common VHL proteins among rat tissues, suggesting tissue- and development-dependent functional difference between these two isoforms. Molecular pathways linking VHL to angiogenesis have been extensively characterized and mechanisms have been proposed to explain how altered VHL leads to tumorigenesis. VHL functions in the presence of oxygen and/or oxygen species. Two strategies are proposed here for anti-tumor and anti-angiogenic treatments of VHL-deficient tumors and those without detectable VHL intragenic mutations. One is to restore wild-type VHL function in VHL-deficient tumors and the other is to enhance wild-type VHL expression and activity in tumors under hypoxic conditions.


Effects of climatic, socio-economic and behavioural factors on the transmission of hookworm (Necator americanus) on two low-country plantations in Sri Lanka.
The climatic, socio-economic and behavioural factors influencing hookworm (Necator americanus) infection in Sri Lanka were explored between February 2000 and June 2001. In February 2000, a single stool sample was collected from each of the 477 subjects investigated, who were aged 2-74 years (median = 13 years) and lived on the 'lowcountry' Maliboda and Ayr plantations. The 'baseline' prevalence (28.5%) and intensity of hookworm infection (0- 4828.5 eggs/g faeces, with a mean of 128.4 eggs/g) were then determined by examining these initial samples, as Kato-Katz smears. Subsequently, each participant was treated with a single, 500-mg dose of mebendazole and then followed-up, at monthly intervals, for the next 15 months. Whenever a subject was found smear-positive for hookworm eggs at one of the monthly follow-ups, he or she was treated again with mebendazole. This approach allowed the monthly incidence of hookworm infection to be determined for each subject, assuming that subjects became smear-positive approximately 6 weeks post-infection. During the study period, rainfall and mean temperature were recorded daily and then converted to monthly values so that the relationship between the incidence of infection over each month and the rainfall and mean temperature over the same period could be explored. In addition, potentially relevant data on the socio-economic status and behaviour of each subject were collected, in questionnaire-based interviews with the adult subjects and the caregivers of the children investigated. Odds ratios (OR) and their 95% confidence intervals (CI) were then calculated for each factor that might increase the risk of hookworm infection. The monthly incidence of hookworm infection showed three peaks -- in September 2000 (21.3%), January 2001 (20.8%) and May 2001 (17.5%) -- at Maliboda, and two peaks -- in September 2000 (25.0%) and February 2001 (29.2%) -- at Ayr. With the data for all subjects combined, incidence showed a statistically significant correlation with mean temperature (r = -0.468; P = 0.018). The results of a multivariate analysis also revealed that mean temperature was significant (beta = -5.296; P = 0.01) in hookworm incidence. Bathing and washing with water from rock-pools formed by waterfalls (OR = 1.33; CI = 1.35-4.01), the use of wells (OR = 2.35; CI = 1.29-4.30), and a lack of toilets (OR = 1.60; CI = 1.01-2.53) each appeared to increase the risk of hookworm infection significantly. Those living on the two study plantations, and perhaps many other similar plantations in Sri Lanka, would clearly benefit from improved access to the public water supply (especially to pipe-borne water) and toilets.


Anaphylaxis in an emergency department: a 2-year study in a tertiary-care hospital.
The aim of this study was to estimate the incidence of anaphylaxis in the emergency department of a tertiary-care hospital, describe the clinical features and the management of the patients and determine those with mild manifestations. A retrospective study was conducted from 2005 to 2006 using anaphylaxis-related ICD-10 terms. Two different sets of criteria for the diagnosis of anaphylaxis were applied, first the criteria previously accepted by emergency practice, followed by the recent criteria set forth at the 2005 international meeting. Sixty-four patients fulfilled the previous criteria with an average incidence of 52.5 per 100,000 patients per year with a shift towards females in 2006. The most common presentations were cutaneous, followed by respiratory symptoms. Food allergy was the most common cause, especially prawn. After applying the recent criteria, 13 patients (20.4%) were excluded, which reduced the incidence to 42.2 per 100,000 patients per year. Long term follow up is suggested for the possible or mild cases that were re-categorized.


Role of facilitators in delivering high quality practice education.
Investment in the professional development of the nursing workforce is essential to enhance patient safety, quality of care, personal development and lifelong learning. When resources are scarce, however, education spending is often the first cost saving and this can reduce quality of care. This article explores the role of education facilitators, who are central to strategic representation of practice education issues, delivery of training and development, determining training needs, and the development of relevant education programmes. In addition, they provide support for mentors, practice educators and clinicians.


Effect of medial opening wedge high tibial osteotomy on intraarticular knee and ankle contact pressures.
High tibial osteotomy (HTO) is a commonly used surgical technique for treating moderate osteoarthritis (OA) of the medial compartment of the knee by shifting the center of force towards the lateral compartment. Previous studies have documented the effects of HTO on the biomechanics of the knee. However, the effects of the procedure on the contact pressures within the ankle joint have not been as well described. Seven cadavers underwent an HTO procedure with sequential 5° valgus realignment of the leg up to 15° of correction. An axial force of up to 550 N was applied and the intraarticular pressure was recorded. Minor valgus realignment of the proximal tibia does not significantly alter the biomechanics of the ankle. However, moderate-to-large changes in proximal tibial alignment result in significantly decreased tibiotalar contact surface area and in changes in intraarticular ankle pressures. These findings are clinically relevant, as they provide a biomechanical rationale for the diagnosis and treatment of ankle symptoms in the setting of lower limb malalignment or after alignment correction procedures.


Methotrexate-induced pulmonary injury: serial CT findings.
We describe serial computed tomographic (CT) findings of methotrexate (MTX)-induced pulmonary injury. The cases of 8 patients (3 men and 5 women; mean age 58.6 years, range 16 to 75 years) of clinically diagnosed MTX-induced pulmonary injury were reviewed. Six patients had rheumatoid arthritis, 1 had lupus erythematosus profundus, and 1 had juvenile rheumatoid arthritis. CT findings on admission and at follow-up were evaluated. The most common CT features were diffuse and patchy bilateral ground-glass opacity with (n = 3) or without reticulation (n = 4) and consolidation (n = 1). These opacities showed no predilection for any particular lung zone in 6 patients but did show dependent predilection in 1 patient and upper lobe predilection in 1. Diffuse centrilobular ill-defined nodules were noted in 1 patient, which disappeared on follow-up. During the average post-treatment follow-up period of 31.0 days (range 3 to 76 days), the opacities quickly improved after treatment in 6 patients; however, in 2 patients with pre-existing interstitial pneumonitis the opacities were refractory. CT features of MTX-induced pulmonary injury were variable and included diffuse parenchymal opacification, reticular opacities, and centrilobular nodules. These opacities usually responded quickly to treatment; however, those patients with lung fibrosis at presentation may have worse prognosis.


Biosafety and biodegradation studies of AZ31B magnesium alloy carotid artery stent in vitro and in vivo.
Biosafety of AZ31B magnesium (Mg) alloy and the effect of its degradation products on tissues, organs, and whole systems are highly needed to be evaluated before clinical application. This study serves a wide variety of safety evaluations of biodegradable AZ31B alloy on nerve cells. As a result of this in vitro study, the maximum aluminum (Al) ion and Mg ion concentrations in the medium were estimated to be 22 μmol/L and 2.75 mmol/L, respectively, during degradation. In addition, the corresponding cell mortality was observed to be 36% and lower than 5% according to the resistance curves of the cell to Mg and Al ions. Furthermore, the maximum Al ion and Mg ion concentrations in serum and cerebrospinal fluid were detected to be 26.1 μmol/L and 1.2 mmol/L, respectively, for 5 months implantation. Combining the result of in vivo dialysis with the result of ion tolerance assay experiments, the actual death rate of nerve cells is estimated between 4 and 10% in vivo, which is lower than the result of in vitro cytotoxicity evaluation. Moreover, no psychomotor disability during clinical studies is observed. Consequently, stent made of AZ31B alloy with surface treatment is feasible for carotid artery stenosis, and it is safe in terms of cell viability on the nervous system.


Molecular cloning of hydroxynitrile lyase from Sorghum bicolor (L.). Homologies to serine carboxypeptidases.
The heterotetrameric enzyme hydroxynitrile lyase (HNL) from sorghum (EC 18.104.22.168) is involved in the catabolism of the cyanogenic glycoside dhurrin. We have isolated a cDNA clone comprising about 90% of the COOH terminal sequence of a precursor which encodes both subunit of HNL from Sorghum bicolor L. (SbHNL). Hence the subunits of SbHNL must be the result of post-translational processing. The deduced amino acid sequence of HNL shares significant sequence homology with members of the serine carboxypeptidase family. In particular, HNL from sorghum shares the catalytical triad Asp. His, and Ser with these enzymes which evolved in 3 groups of enzymes (carboxypeptidase, chymotrypsin, and subtilisin) by convergent evolution. Moreover, like serine carboxypeptidases, HNL from sorghum consists of two pairs of glycosylated cysteine linked A and B chains forming a heterotetramer of a molecular weight of 105,000 (carboxypeptidases 120,000). Thus, HNL from sorghum closely resembles to serine carboxypeptidases but differs from all other HNLs described so far. Western blotting experiments revealed cross reaction between carboxypeptidase from wheat and anti SbHNL antisera. Therefore, convergent evolution of HNLs from various ancestoral enzymes is conceivable. Hybridization of SbHNL cDNA to northern blots of total RNAs isolated from various organs of young sorghum seedlings shows the same expression pattern of HNL as found by means of western blotting or enzyme assays. Using PCR and Southern blot analysis, we demonstrated that the gene of SbHNL is free of introns. Further sequence analysis of cDNA clones and genomic DNA revealed a stretch of 23 adenine residues in the 3'-untranslated part of the gene. Both, intronless organisation of the gene and a genomic stretch of oligo A suggests that SbHNL may have evolved by a reverse transcription event.


A simple matrine derivative for the facile syntheses of mesoporous zeolites ITQ-37 and ITQ-43.
ITQ-37 and ITQ-43 possessing mesoporous structures have aroused great interest because of their promising applications involving large molecules. However, it is challenging to synthesize these zeolites. Herein, MAS-ITQ-37 and MAS-ITQ-43 with enhanced Si/Ge ratios of 1.7 and 3.0 have been synthesized by using a simple matrine-derived organic structure-directing agent.


Subjective visual vertical in vestibular disorders measured with the bucket test.
The 'bucket test' may indicate that patients with known vestibular disorders have spatial orientation deficits but due to the low receiver operating characteristics (ROC) values it is not useful for screening people for vestibular impairments. 1) to verify that patients with unilateral peripheral vestibular weakness (UW) differ from normal subjects on the bucket test, 2) to determine if patients with unilateral benign paroxysmal positional vertigo (BPPV) differ from normal subjects, 3) to determine the sensitivity and specificity of the test. Patients with UW (n = 25) and BPPV (n = 25) were compared to normals (n = 50). Subjects looked into a clean bucket with a vertical line on the bottom, which rested on a table. It was rotated, clockwise or counterclockwise, for three trials per direction until the subject indicated that the line was vertical. The dependent measure was the mean absolute value of the deviations from the true vertical. Some, but not all, patients' responses differed from normal subjects but responses also differed by age and sex. ROC values were all weak, i.e. < 0.8. No good cut-off points differentiated controls from patients. Thus, although the bucket test is useful for describing spatial deficits in patients this test is not useful for screening people for possible vestibular impairments.


Paediatric burn injuries requiring hospitalization in Fars, Southern Iran.
The aim of this study was to determine the demographic characteristics of burnt children in need of hospitalization, causes of burns and associated complications in Fars province, Iran. This is a retrospective cross-sectional study. Files of all children under 15 years of age who were hospitalized in the only burn referral centre of Fars province were evaluated. Data regarding age, gender, location (urban, rural), burn surface area (BSA), cause of burn, length of hospital stay and complications were extracted from patients' files. Data were analyzed using SPSS, version 22 and the Chi-square test. A p-value of less than 0.05 was considered as statistically significant. A total of 122 patients (54.9% males) were studied. Children from rural areas were hospitalized 1.4 times more often than urban children. Overall, 31.2% of admissions occurred in winter. Burning with hot liquids (scalding) was the most common cause of burns (56.6%, n = 69). Mean BSA was 12.29 ± 21.18% and mean length of hospital stay was 7.59 ± 12.78 days. Burn complications were seen in 19 cases (15.6%). One child died due to inhalational thermal injury. This study showed that burns mostly occur in boys, in the winter and in rural areas of Fars province. Furthermore, scald burns are the most common type of burn injury. Since a significant number of children suffer from permanent complications following burn injuries, special planning is needed to prevent this type of injury.


Cochlear implantation in children: the problem of head growth.
Successful implantation in children requires that provision be made for later head growth. The timing and magnitude of this growth was determined by measurements made from high-resolution computerized tomographic images of developing temporal bones. The temporal bone scans of 103 children of varying ages were studied. The development of the petrous and squamous portion of the temporal bone was at its greatest in the first 2 years of life. The order of magnitude of growth varied with the different axes measured.


[Integrated and automated data analysis for neuronal activation studies using positron emission tomography: methodology and applications].
A data analysis method was developed for neuronal activation studies using [15O]water positron emission tomography (PET). The method consists of several procedures including intra-subject head motion correction (co-registration), detection of the mid-sagittal plane of the brain, detection of the intercommissural (AC-PC) line, linear scaling and non-linear warping for anatomical standardization, pixel-by-piexl statistical analysis, and data display. All steps are performed in three dimensions and are fully automated. Each step was validated using a brain phantom, computer simulations, and data from human subjects, demonstrating accuracy and reliability of the procedure. The method was applied to human neuronal activation studies using vibratory and visual stimulations. The method detected significant blood flow increases in the primary sensory cortices as well as in other regions such as the secondary sensory cortex and cerebellum. The proposed method should enhance application of PET neuronal activation studies to the investigation of higher-order human brain functions.


Takotsubo Cardiomyopathy (Broken-Heart Syndrome): A Short Review.
Takotsubo cardiomyopathy, also called "broken heart" syndrome or apical ballooning syndrome, is a reversible cardiomyopathy characterized by left ventricular dysfunction and ballooning of the left ventricular apex on imaging during systole. It predominantly occurs in post-menopausal women and is commonly associated with emotional or physical stress. Patients commonly present with chest pain and electrocardiographic evidence of ST segment elevation or T-wave-mimicking acute coronary syndrome, but with an absence of angiographic evidence of obstructive coronary disease. The exact cause is unknown, but potential contributors include catecholamine excess and sympathetic nervous system hyperactivity. There is no consensus on pharmacological treatment of takotsubo cardiomyopathy. Based on the suspected pathophysiology of the disease, adrenergic blockade using beta-blocker therapy is employed. Near complete resolution of left ventricular wall motion dyskinesis occurs in the majority of takotsubo cardiomyopathy patients within a month. Although the prognosis is generally favorable, there are reports of complications during the acute phase, including cardiogenic shock, pulmonary edema, ventricular tachycardia, apical thrombus formation, and death. This review article will briefly discuss the epidemiology, etiology, clinical features, diagnostic evaluation, and treatment of this condition.


Identification of [(naphthalene-1-carbonyl)-amino]-acetic acid derivatives as nonnucleoside inhibitors of HCV NS5B RNA dependent RNA polymerase.
A novel series of HCV NS5B RNA dependent RNA polymerase inhibitors containing a naphthalene carboxamide scaffold were identified by high throughput screening. Optimization of substituents by parallel synthesis and the iterative design towards understanding structure-activity relationship to improve potency are described. Tetra substituted naphthalene 31 displayed potent activity with IC(50) of 120 nM against HCV NS5B enzyme and was selective over a panel of polymerases.


Bioaugmentaton in Technosols created in abandoned pyritic tailings can contribute to enhance soil C sequestration and plant colonization.
Creation of Technosols in combination with phytostabilization may be a sustainable strategy to minimize the environmental and human health hazards derived from mine tailings. Bioaugmentation can facilitate plant establishment and growth for efficient phytostabilization. In order to assess if bioaugmentation can increase soil quality and fertility, decrease metal(loid) mobility and accelerate plant colonization, a one-year field experiment was designed with creation of Technosols in two tailings ponds with different pH (acidic (AT) and neutral (NT)), with addition of marble waste (MaW) and two organic materials (pig manure (PM) and sewage sludge (SS), without or with inoculation of effective microorganisms (EM) at three rates. Results showed that MaW was the main factor responsible for maintaining pH ~7 in AT and ~8 in NT, decreasing salinity, and decreasing the soluble fraction of metals (70-99%). The soluble fraction of As decreased ~45% in AT, related to increases in pH up to neutrality, while increased ~90% in NT with PM due to pH>8 and higher content of organic compounds. The addition of PM and SS significantly increased soil organic C (SOC), nutrient contents and microbial biomass and activity in both tailings, being PM more effective. However, a positive priming effect was observed in NT with SS addition likely due to higher C/N ratio and lack of nutrients. There was a significant effect of EM rate on inorganic C, SOC, N, K and microbial biomass and activity, with higher values as rate increased. Vegetation richness and density directly increased with increasing EM rate. Multivariate analyses showed that the most important properties contributing to increase richness and plant density were microbial biomass and N. Thus, bioaugmentation contributed to soil C sequestration (as organic and inorganic C) and soil fertility, related to high soil microbial biomass and activity, which facilitated an effective colonization of vegetation.


Pathologically Proven Spontaneous Remission of IgG4-related Retroperitoneal Fibrosis.
Some forms of idiopathic retroperitoneal fibrosis (RF) have recently been considered to be a part of the spectrum of immunoglobulin G4 (IgG4)-related disease. This case report is the first description of a spontaneous remission in a patient with pathologically proven IgG4-related RF. Although the pathogenesis and long-term disease behavior of IgG4-related RF remains unknown, we believe that an initial assessment consisting of only careful monitoring might be one important strategy, especially in asymptomatic IgG4-related RF patients without nephropathy, while carefully monitoring these patients for the risk of recurrence.


Use of quality of life measures in clinical trials.
To be useful in clinical trials, quality of life (QOL) measures must meet basic criteria related to psychometric properties (reliability, validity, and sensitivity), length and acceptability to patients, and availability of parallel child and proxy versions. This paper examines the adequacy of several current measures, with special reference to childhood cancer. We identified 10 such measures of varying length, age-appropriateness, and content. We also discerned a number of obstacles to inclusion of QOL measures in trials. These include clinician bias, financial cost, and time required to inform families and to obtain informed consent from parents. We noted some methodological difficulties associated with measuring QOL over time. These include allowing for maturation and accounting for response shift, or children's propensity to change their internal standards for assessing QOL.


[Antegrade puncture of the superficial femoral artery: a pilot project].
Antegrade puncture of the femoral artery is often the first step in angioplasty or thrombolysis of the lower limb. Because of presumed complications, mostly from anecdotal literature, the common femoral artery is the preferred site of entry. From there, selective catheterization of the superficial femoral artery can be performed. This two-step procedure is sometimes long and difficult for both the patient and the radiologist. To improve the ease and the rapidity of this procedure, the author performed direct antegrade puncture of the superficial femoral artery during 25 consecutive antegrade approaches. The only local complication was hematoma, which developed in two patients but disappeared spontaneously. Aggravating factors were the size of the catheters used and fibrinolytic therapy in one of the affected patients. The results of this pilot study suggest strongly that direct antegrade puncture of the superficial femoral artery is safer than might be expected from the literature, which deals primarily with retrograde puncture. This approach must now be systematically compared with the "classic" common femoral artery approach.


Ultrastructural evidence for association of vascular dendritic cells with T-lymphocytes and with B-cells in human atherosclerosis.
It has been previously demonstrated that dendritic cells are involved in human atherogenesis and immunohistochemical analysis indicated that S100+/CD1a+ vascular dendritic cells co-localize with lymphocytes in atherosclerotic lesions. Despite the high frequency of co-localization of vascular dendritic cells and lymphocytes in cell-rich areas, different type intimal cells were often intermingled and very closely apposed, which precluded determining whether vascular dendritic cells and lymphocytes specifically contacted each other or were simply densely packed together. Therefore, we undertook the present electronmicroscopical examination to see if vascular dendritic cells form direct contacts with lymphocytes in human atherosclerotic lesions. Serial sectioning of lymphocyte-rich areas of the human atherosclerotic aortas showed that vascular dendritic cells exhibiting a well developed tubulovesicular apparatus were located amongst T-lymphocytes and that through their processes, vascular dendritic cells formed multiple contacts with T-lymphocytes. In these contacts, vascular dendritic cell processes exhibited hypertrophied cisterns of the tubulovesicular system, which suggests the activation of vascular dendritic cells. From our observations we speculate that vascular dendritic cells may be responsible for the survival and activation of T-lymphocytes since a large number of T-lymphocytes which did not form contacts with vascular dendritic cells were found to have undergone destruction in atherosclerotic lesions. B-cells (plasma cells) were rarely seen in atherosclerotic lesions but vascular dendritic cells were found to form direct contacts with them. That vascular dendritic cells contact with both the T- and B-cells suggests that these vascular dendritic cells differ from other dendritic cells, subtypes of which are associated with T-cells (Langerhans cells, interdigitating cells) or with B-cells (follicular dendritic cells).


Direct indicators of social distancing effectiveness in COVID-19 outbreak stages: a correlational analysis of case contacts and population mobility.
The effectiveness of social distancing during the coronavirus disease 2019 (COVID-19) pandemic has been evaluated using the magnitude of changes in population mobility. This study aimed to investigate a direct indicator-namely, the number of close contacts per patient with confirmed COVID-19. From week 7, 2020 to week 43, 2021, population movement changes were calculated from the data of two Korean telecommunication companies and Google in accordance with social distancing stringency levels. Data on confirmed cases and their close contacts among residents of Gyeonggi Province, Republic of Korea were combined at each stage. Pearson correlation analysis was conducted to compare the movement data with the change in the number of contacts for each confirmed case calculated by stratification according to age group. The reference value of the population movement data was set using the value before mid-February 2020, considering each data's characteristics. In the age group of 18 or younger, the number of close contacts per confirmed case decreased or increased when the stringency level was strengthened or relaxed, respectively. In adults, the correlation was relatively low, with no correlation between the change in the number of close contacts per confirmed case and the change in population movement after the commencement of vaccination for adults. The effectiveness of governmental social distancing policies against COVID-19 can be evaluated using the number of close contacts per confirmed case as a direct indicator, especially for each age group. Such an analysis can facilitate policy changes for specific groups.


PP-1β and PP-2Aα modulate cAMP response element-binding protein (CREB) functions in aging control and stress response through de-regulation of αB-crystallin gene and p300-p53 signaling axis.
The function of the transcription factor, cAMP response element-binding protein (CREB), is activated through S133 phosphorylation by PKA and others. Regarding its inactivation, it is not well defined. cAMP response element-binding protein plays an essential role in promoting cell proliferation, neuronal survival and the synaptic plasticity associated with long-term memory. Our recent studies have shown that CREB is an important player in mediating stress response. Here, we have demonstrated that CREB regulates aging process through suppression of αB-crystallin and activation of the p300-p53-Bak/Bax signaling axis. First, we determined that two specific protein phosphatases, PP-1β and PP-2Aα, can inactivate CREB through S133 dephosphorylation. Subsequently, we demonstrated that cells expressing the S133A-CREB, a mutant mimicking constant dephosphorylation at S133, suppress CREB functions in aging control and stress response. Mechanistically, S133A-CREB not only significantly suppresses CREB control of αB-crystallin gene, but also represses CREB-mediated activation of p53 acetylation and downstream Bak/Bax genes. cAMP response element-binding protein suppression of αB-crystallin and its activation of p53 acetylation are major molecular events observed in human cataractous lenses of different age groups. Together, our results demonstrate that PP-1β and PP-2Aα modulate CREB functions in aging control and stress response through de-regulation of αB-crystallin gene and p300-p53-Bax/Bak signaling axis, which regulates human cataractogenesis in the aging lens.


Localization and analyses of small drug molecules in rat brain tissue sections.
Traditional detection of drugs in tissue requires tissue homogenization, which precludes the mapping and localization of drugs. The use of autoradiography could compensate for such shortcoming. However, it requires expensive custom-synthesized radioactive drugs. Recent improvement in sample preparation for matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) and MALDI-MS/MS provides an alternative approach for in situ drug detection. In this work, rat brains were collected after intracranial injection of chlorisondamine or intraperitoneal injection of cocaine and snap frozen. MALDI matrixes were applied directly to 14-mum brain cryosections and spectra acquired. The identity of the drugs was further confirmed by MS/MS. Careful matrix selection and tissue preparation allows for the successful detection of drugs and the mapping of their relative abundance across various regions of the brain. This new method is simple, safe, accurate, fast, cost-effective, and low in sample consumption and shows potential for imaging, pharmacokinetics, and toxicology applications.


Network Pharmacology-Based Approach to Investigate the Active Ingredients and Therapeutic Mechanisms of Jingu Tongxiao Pill against Osteoarthritis.
This study aimed to investigate the active ingredients and therapeutic mechanisms of Jingu Tongxiao Pill (JGTXP), a commonly used Chinese patent medicine, in treating osteoarthritis (OA) via network pharmacology analysis combined with experimental validation. First, we administered JGTXP to rat plasma and identified the candidate active compounds. Next, target prediction, protein-protein interaction, compound-target network construction, gene ontology (GO), and Kyoto Encyclopedia of Genes and Genomes pathway enrichment analyses were conducted for JGTXP. Lastly, the network-derived key targets and pathways were validated in vitro and in vivo. Finally, we identified 106 compounds in JGTXP and 24 absorbed compounds in the rat plasma. Network analysis revealed that JGTXP interferes with OA mainly via regulating the inflammatory response, collagen catabolic process, and osteoclast differentiation, and the nuclear factor kappa B (NF-κB) signaling pathway plays a pivotal role in these processes. Experimentally, JGTXP exerted potential protective effects on articular cartilage and inhibited expression of inflammatory mediators and collagen catabolism-related proteins, including interleukin 1 beta (IL-1β), interleukin 6, tumor necrosis factor alpha (TNF-α), and matrix metalloproteinase (MMP) 3 and MMP13, in a papain-induced OA rat model. Consistently, mRNA expression levels of these factors and nitric oxide release were suppressed by JGTXP in an LPS-induced RAW 264.7 inflammation model. The reporter gene assay showed that JGTXP could reduce the transcriptional activity of NF-κB. Consecutive western blot analysis demonstrated that nuclear NF-κB p65, inducible nitric oxide synthase (iNOS), and cyclooxygenase 2 (COX-2) expression were inhibited while cytoplasmic NF-κB p65 was upregulated by JGTXP. Using a combination of chemical profiling, network pharmacology analysis, and experimental validation, we preliminarily clarified the active ingredients of JGTXP intervention for OA and demonstrated that JGTXP ameliorates OA, at least partially, by regulating the NF-κB signaling pathway.


Phylogenetic placement of the anamorphic tribe Ustilaginoideae (Hypocreales, Ascomycota).
Tribe Ustilaginoideae (Hypocreales, Ascomycetes) is made up of three anamorph genera, Munkia, Neomunkia and Ustilaginoidea. Species of Munkia and Neomunkia develop on the culms of bamboo (Chusquea spp.) and have a neotropical distribution while species of Ustilaginoidea infect the florets of various grasses and are pantropical in distribution. In this study we evaluated the phylogeny of the tribe and assessed hypotheses regarding its affinity to clavicipitalean teleomorphic groups. To support phylogenetic analyses, morphology of representatives of several key species of Ustilaginoideae was examined also. Phylogenetic analyses using sequences of the large subunit of the ribosomal RNA gene suggest that members of Ustilaginoideae are distinct from teleomorphic genera of Clavicipitaceae and that Ustilaginoideae should be recognized as a monophyletic group within Hypocreales. However, phylogenetic analyses did not resolve the placement of Ustilaginoideae in Clavicipitaceae or Hypocreaceae, suggesting that it might be a distinct lineage within Hypocreales. This evaluation supported the monophyly of tribes Balansieae and Clavicipeae in the family Clavicipitaceae.


Intracellular product recycling in high succinic acid producing yeast at low pH.
The metabolic engineering of Saccharomyces cerevisiae for the production of succinic acid has progressed dramatically, and a series of high-producing hosts are available. At low cultivation pH and high titers, the product transport can become bidirectional, i.e. the acid is reentering the cell and is again exported or even catabolized. Here, a quantitative approach for the identification of product recycling fluxes is developed. The metabolic flux distributions at two time-points of the fermentation process were analyzed. 13C labeled succinic acid was added to the extracellular space and intracellular enrichments were measured and subsequently used for the estimation of metabolic fluxes. The labeling was introduced by a labeling switch experiment, leading to an immediate labeling of about 85% of the acid while keeping the total acid concentration constant. Within 100 s significant labeling enrichment of the TCA cycle intermediates fumarate, iso-citrate and α-ketoglutarate was observed, while no labeling was detected for malate and citrate. These findings suggest that succinic acid is rapidly exchanged over the cellular membrane and enters the oxidative TCA cycle. Remarkably, in the oxidative direction malate 13C enrichment was not detected, indicating that there is no flux going through this metabolite pool. Using flux modeling and thermodynamic assumptions on compartmentation it was concluded that malate must be predominantly cytosolic while fumarate and iso-citrate were more dominant in the mitochondria. Adding labeled product without changing the extracellular environment allowed to quantify intracellular metabolic fluxes under high producing conditions and identify product degradation cycles. In the specific case of succinic acid production, compartmentation was found to play a major role, i.e. the presence of metabolic activity in two different cellular compartments lead to intracellular product degradation reducing the yield. We also observed that the flux from glucose to succinic acid branches at two points in metabolism: (1) At the level of pyruvate, and (2) at cytosolic malate which was not expected.


Does birth weight predict childhood diet in the Avon longitudinal study of parents and children?
Low birth weight predicts cardiovascular disease in adulthood, and one possible explanation is that children with lower birth weight consume more fat than those born heavier. Therefore, the objective of this study was to investigate associations between birth weight and childhood diet, and in particular, to test the hypothesis that birth weight is inversely related to total and saturated fat intake. Prospective cohort study. South west England. A subgroup of children enrolled in the Avon longitudinal study of parents and children, with data on birth weight and also diet at ages 8, 18, 43 months, and 7 years (1152, 998, 848, and 771 children respectively). Associations between birth weight and diet increased in strength from age 8 to 43 months, but had diminished by age 7 years. Fat, saturated fat, and protein intakes were inversely, and carbohydrate intake was positively associated with birth weight at 43 months of age, after adjusting for age, sex, and energy intake. After adjustment for other confounders, all associations were weakened, although there was still a suggestion of a relation with saturated fat (-0.48 (95% CI -0.97, 0.02) g/day per 500 g increase in birth weight. Similar patterns were seen in boys and girls separately, and when the sample was restricted to those with complete data at all ages. A small inverse association was found between birth weight and saturated fat intake in children at 43 months of age but this was not present at 7 years of age. This study therefore provides little evidence that birth weight modifies subsequent childhood diet.


Development of dendritic trees of rabbit retinal alpha ganglion cells: relation to differential retinal growth.
To provide a quantitative description of the postnatal development of dendritic trees in alpha ganglion cells of the rabbit retina, these cells were stained either by intracellular injection of Lucifer yellow or by application of the lipophilic dye DiI. This was done at three developmental stages: postnatal day (P) 8/9, P 16/17, and in adults. For different retinal locations we quantified the alpha cell dendritic field area, the number of dendritic branch points, and the average dendritic length between branch points. According to the alpha cell location, the data were collected in three groups representing the retinal center, midperiphery, and far periphery, respectively. The data were then correlated with the postnatal retinal expansion which is known to differ among the above topographic regions of the retinae (Reichenbach et al., 1993). Our results show that the growth of alpha ganglion cell dendrites is not proportional to, but significantly exceeds, that of the local retinal tissue. Between P 8/9 and adulthood, the area of central alpha cells increases almost six-fold from 26,000 to 144,000 microns 2 (retinal expansion: 2.2-fold), and that of peripheral cells more than 15-fold from 35,000 to 556,000 microns 2 (retinal expansion: four-fold). During this period, the coverage factor of alpha cell dendritic fields increases about three-fold, and reaches adult levels of about 3 (retinal center) and 2.2 (periphery), respectively. The number of dendritic branch points remains nearly constant, and the distance between them increases by a factor close to the square root of the factor by which the dendritic field area grows. Thus, it appears that, from the second postnatal week on, dendritic trees of rabbit alpha ganglion cells increase by intense "interstitial growth," rather than by outgrowth of (new) dendritic branches. This growth pattern is different from that of some other rabbit retinal ganglion cell types, and of alpha ganglion cells of the cat retina, whose dendritic trees expand at a rate equal to or less than that of the surrounding retinal tissue. The consequences for synaptic contacts with bipolar and amacrine cells are discussed; they suggest a high degree of synaptic plasticity during normal postnatal retinal growth.


Use of a marginal structural model to determine the effect of aspirin on cardiovascular mortality in the Physicians' Health Study.
The 1982-1988 aspirin component of the Physicians' Health Study, a randomized trial of aspirin and beta-carotene in primary prevention of cardiovascular disease and cancer among 22,071 US male physicians, was terminated early primarily because of a statistically extreme 44% reduction in first myocardial infarction, with inadequate precision and no apparent effect on the primary endpoint, cardiovascular death. Because of the demonstrated efficacy of aspirin in secondary prevention of cardiovascular death, nonfatal cardiovascular events may simultaneously be time-dependent confounders and intermediate variables. Aspirin use is strongly influenced by these as well as other diseases, side effects, and cardiovascular risk factors. The authors used a marginal structural model with time-dependent inverse probability weights to estimate the underlying causal effect of aspirin on cardiovascular mortality. Although intention-to-treat analyses found no effect (rate ratio = 1.00, 95% confidence interval (CI): 0.72, 1.38), the estimated causal rate ratio was altered to 0.75 but remained nonsignificant (95% CI: 0.48, 1.16). As-treated analyses suggested a more modest effect of aspirin use (rate ratio = 0.90, 95% CI: 0.65, 1.25). Although the numbers of cardiovascular deaths were insufficient to evaluate this endpoint definitively, use of such methods holds much potential for controlling time-varying confounders affected by previous exposure.