Up-regulation of stem cell markers by P21-activated kinase 1 contributes to 5-fluorouracil resistance of colorectal cancer.
Cancer stem cells (CSC) are tumorigenic and resistant to chemotherapy. In colorectal cancer (CRC), CSCs have been identified by the expression of specific markers, including CD44, Bmi1 and Nanog. Although p21-activated kinase 1 (PAK1), acting downstream of Ras, stimulates Wnt/β-catenin signaling and is known to play an important role in CRC development and progression, the role of PAK1 in the expression of CSC markers has not previously been investigated. The effect of PAK1 over-expression, knockdown or inhibition on the expression or alteration (in the case of CD44) of CSC markers in human CRC cell lines was measured by immunofluorescence and Western blotting. The effect of PAK1 modulation on tumorigenesis, and on resistance to treatment with 5-fluorouracil (5-FU), was measured by sphere formation in vitro and by growth of xenografted tumors in vivo. The results show that PAK1 activity correlated with the expression of CSC markers and the CD44 isoform profile, and with tumor growth both in vitro and in vivo. Furthermore PAK overexpression partially overcame the inhibition of CRC growth by 5-FU, and PAK inhibition was synergistic with 5-FU treatment. Our findings lay the foundation for a combination therapy in which PAK1 inhibitors targeting CSCs may be combined with conventional 5-FU-based chemotherapy for the treatment of CRC.


Conformationally constrained [p-(omega-aminoalkyl)phenacetyl]-L-seryl-L-lysyl dipeptide amides as potent peptidomimetic inhibitors of Candida albicans and human myristoyl-CoA:protein N-myristoyl transferase.
MyristoylCoA:protein N-myristoyltransferase (NMT) covalently attaches the 14-carbon saturated fatty acid myristate, via an amide bond, to the N-terminal glycine residues of a variety of cellular proteins. Genetic studies have shown that NMT is essential for the viability of the principal fungal pathogens which cause systemic infection in immunosuppressed humans and hence is a target for development of fungicidal drugs. We have generated a class of potent peptidomimetic inhibitors of the NMT from one such fungal pathogen, Candida albicans. The N-terminal tetrapeptide from a substrate analog inhibitor, ALYASKL-NH2, was replaced with an omega-aminoalkanoyl moiety having an optimal 11-carbon chain for inhibition (11-aminoundecanoyl-SKL-NH2, 3a, IC50 = 1.2 +/- 0.14 microM). A series of replacements for the C-terminal Leu established that residues containing a lipophilic side chain were most effective, with cyclohexylalanine having the greatest potency (3g, IC50 = 0.36 +/- 0.06 microM). Removal of the carboxamide moiety led to a metabolically stable dipeptide inhibitor containing an N-(cyclohexylethyl)lysinamide (17e, IC50 = 0.11 +/- 0.03 microM). Partial rigidification of the flexible aminoundecanoyl chain produced the dipeptide p-(omega-aminohexyl)phenacetyl-L-seryl-L-lysyl-N-(cyclohexyleth yl)amide (26b, IC50 = 0.11 +/- 0.04 microM). Subsequent incorporation of an alpha-methyl substituent into 26b provided the dipeptide analog [2-[p-(omega-aminohexyl)phenyl]propionyl]-L-seryl-L-lysyl-N-(cyclohex ylethyl)amide, a very potent inhibitor (48, IC50 = 0.043 +/- 0.006 microM), which retained the three essential elements required for recognition by the acyl transferase's peptide binding site.


Pathogenesis of Trypanosoma brucei infection in deer mice (Peromyscus maniculatus). Light and electron microscopic study of testicular lesions.
The results of light and electron microscopic studies demonstrated that Trypanosoma brucei infection produced marked testicular degeneration in deer mice (Peromyscus maniculatus) with highly significant (p less than 0.001) decreases in testicular weight, seminiferous tubular diameter, and epithelial thickness. Lesions in seminiferous tubules consisted of necrosis of spermatids, spermatocytes, and sometimes the spermatogonia. Sertoli's cells had marked vacuolation and accumulation of phagocytic material. Orchitis was present in most infected mice, particularly those with severe seminiferous tubular degeneration. There was marked folding of the basal laminae of the seminiferous tubule and myoid layer, and a consistent increase in the layers of the basal laminae of the seminiferous tubule. Trypanosomes were present in the intertubular tissues of seminiferous tubules, and they occasionally crossed the myoid layer but never the basal lamina of seminiferous tubules. Inflammatory and other cells in the interstitium consisted of lymphocytes (35.2%), Leydig's cells (25.1%), macrophages (18.1%), plasma cells (12.3%), neutrophils (5.6%), eosinophils (3.2%), and mast cells (0.5%). Inflammatory cells including lymphocytes (51.6%), macrophages (43.9%), plasma cells (3.0%), and eosinophils (1.5%) occasionally breached the myoid layer and occupied the space between it and the basal lamina of the seminiferous tubule. Leydig's cells usually existed in clusters, had decreased mitochondrial size and secretory granules, and folding of the nuclear membrane.


Associative symmetry versus independent associations in the memory for object-location associations.
The formation of associations between objects and locations is a vital aspect of episodic memory. More specifically, remembering the location where one experienced an object and, vice versa, the object one encountered at a specific location are both important elements for the memory of an event. Whether episodic associations are holistic representations of individual components or whether there are unidirectional, separately modifiable connections between them has been investigated nearly exclusively using verbal stimuli. A preliminary conclusion concerning this controversy is that verbal associations are, at least, highly correlated (M. J. Kahana, 2002). This theoretical debate, which in the past has undergone a major empirical effort, is still of relevance for the concurrent global matching models of associative memory (S. E. Clark & S. D. Gronlund, 1996). The authors used variations of a novel object-location learning paradigm to complement the accumulated evidence regarding the nature of episodic associations.


Providing Psychotherapy in an Urban, Underserved Community during the COVID-19 Pandemic.
Psychologists housed in two family medicine residency clinics located in underserved communities quickly transitioned their mode of psychotherapy from in-person to completely virtual at the beginning of the COVID-19 pandemic. We share numerous successes and advantages of a telehealth model in serving our community and describe challenges we have encountered.


Investigation of Medium Chain Fatty Acid Feed Supplementation for Reducing Salmonella Typhimurium Colonization in Turkey Poults.
Studies indicate that persistent Salmonella colonization occurs in poultry that are infected early in life, leading to both food safety and public health concerns. Development of improved preharvest Salmonella management strategies is needed to reduce poultry product contamination. The objective of this study was to evaluate the efficacy of a product containing medium chain fatty acids (MCFA) for reducing early Salmonella colonization in turkey poults. Day-of-hatch turkeys were provided a standard starter diet supplemented with MCFA at 0 (negative and positive controls), 1.5, 3, 4.5, or 6 lbs/ton of feed. Positive control and MCFA treated birds were also crop-gavaged with 108 colony forming units (CFU) of bioluminescent Salmonella Typhimurium. Gastrointestinal tissue samples were collected at 3 days postinoculation for bioluminescence imaging (Meckel's diverticulum to the cloaca) and selective enumeration (cecal contents). Quantification of bioluminescence indicated that the 4.5 and 6 lbs/ton MCFA groups had significantly less colonization than the positive control group (p = 0.0412 and p < 0.0001, respectively). Similarly, significantly lower numbers (1-log10 CFU/g reduction) of Salmonella were observed in the ceca of the 6 lbs/ton MCFA group compared to the positive control group (p = 0.0153). These findings indicate that incorporation of MCFA in turkey diets can significantly reduce early Salmonella colonization. In addition, this study highlights the utility of bioluminescence imaging as a screening methodology for assessing the efficacy of treatments that may reduce Salmonella in poultry.


Trial of labor after cesarean delivery for pregnancies complicated by gestational diabetes mellitus.
To assess rates of, and factors associated with, trial of labor and vaginal delivery among pregnancies complicated by both gestational diabetes (GDM) and a previous cesarean delivery. In a retrospective study of singleton deliveries at a university hospital in Israel between 2009 and 2015, women with one previous cesarean delivery and GDM were matched to those without GDM by past cesarean delivery indication: non-favorable (non-progressive labor or failed induction) or favorable (all other indications). Computerized files were reviewed, and maternal, obstetric, and neonatal outcomes were compared. In total, 109 women with GDM were matched to 109 control women. Trial of labor was attempted by 55 (50.5%) women in the control group and 11 (10.1%) in the GDM group (P<0.001). Overall, 42 (38.5%) control women and 5 (4.6%) women with GDM delivered vaginally (P<0.001). Multivariable logistic regression identified lack of GDM as the most significant factor associated with trial of vaginal delivery, followed by a prior vaginal delivery. Among women attempting a trial of labor, past vaginal delivery, but not GDM status, was associated with successful vaginal delivery. GDM significantly diminished the likelihood of a trial of vaginal delivery after cesarean delivery and its subsequent success.


Development of a Rubredoxin-Type Center Embedded in a de Dovo-Designed Three-Helix Bundle.
Protein design is a powerful tool for interrogating the basic requirements for the function of a metal site in a way that allows for the selective incorporation of elements that are important for function. Rubredoxins are small electron transfer proteins with a reduction potential centered near 0 mV (vs normal hydrogen electrode). All previous attempts to design a rubredoxin site have focused on incorporating the canonical CXXC motifs in addition to reproducing the peptide fold or using flexible loop regions to define the morphology of the site. We have produced a rubredoxin site in an utterly different fold, a three-helix bundle. The spectra of this construct mimic the ultraviolet-visible, Mössbauer, electron paramagnetic resonance, and magnetic circular dichroism spectra of native rubredoxin. Furthermore, the measured reduction potential suggests that this rubredoxin analogue could function similarly. Thus, we have shown that an α-helical scaffold sustains a rubredoxin site that can cycle with the desired potential between the Fe(II) and Fe(III) states and reproduces the spectroscopic characteristics of this electron transport protein without requiring the classic rubredoxin protein fold.


Atom-counting in High Resolution Electron Microscopy:TEM or STEM - That's the question.
In this work, a recently developed quantitative approach based on the principles of detection theory is used in order to determine the possibilities and limitations of High Resolution Scanning Transmission Electron Microscopy (HR STEM) and HR TEM for atom-counting. So far, HR STEM has been shown to be an appropriate imaging mode to count the number of atoms in a projected atomic column. Recently, it has been demonstrated that HR TEM, when using negative spherical aberration imaging, is suitable for atom-counting as well. The capabilities of both imaging techniques are investigated and compared using the probability of error as a criterion. It is shown that for the same incoming electron dose, HR STEM outperforms HR TEM under common practice standards, i.e. when the decision is based on the probability function of the peak intensities in HR TEM and of the scattering cross-sections in HR STEM. If the atom-counting decision is based on the joint probability function of the image pixel values, the dependence of all image pixel intensities as a function of thickness should be known accurately. Under this assumption, the probability of error may decrease significantly for atom-counting in HR TEM and may, in theory, become lower as compared to HR STEM under the predicted optimal experimental settings. However, the commonly used standard for atom-counting in HR STEM leads to a high performance and has been shown to work in practice.


Does Iodine Intake Modify the Effect of Maternal Dysglycemia on Birth Weight in Mild-to-Moderate Iodine-Deficient Populations? A Mother-Newborn Prospective Cohort Study.
It is unclear how maternal glycemic status and maternal iodine status influence birth weight among individuals with mild-to-moderate iodine deficiency (ID). We studied the association between birth weight and both maternal glucose levels and iodine intake among pregnant women with mild-to-moderate ID. Glucose values were assessed using a glucose challenge test (GCT) and non-fasting glucose levels that were determined before delivery; individuals' iodine statuses were assessed using an iodine food frequency questionnaire; and serum thyroglobulin (Tg) and urinary iodine concentrations (UIC) were used to assess each group's iodine status. Thyroid antibodies and free thyroxine (FT4) levels were measured. Obstetric and anthropometric data were also collected. Large-for-gestational age (LGA) status was predicted using a Cox proportional hazards model with multiple confounders. Tg > 13 g/L was independently associated with LGA (adjusted hazard ratio = 3.4, 95% CI: 1.4-10.2, p = 0.001). Estimated iodine intake correlated with FT4 among participants who reported consuming iodine-containing supplements (ICS) after adjusting for confounders (β = 0.4, 95% CI: 0.0002-0.0008, p = 0.001). Newborn weight percentiles were inversely correlated with maternal FT4 values (β = -0.2 95% CI:-0.08--56.49, p = 0.049). We conclude that in mild-to-moderate ID regions, insufficient maternal iodine status may increase LGA risk. Iodine status and ICS intake may modify the effect that maternal dysglycemia has on offspring weight.


Invasive group A streptococcal infection concurrent with 2009 H1N1 influenza.
We describe 10 patients with 2009 H1N1 influenza and concurrent invasive group A streptococcal infection with marked associated morbidity and mortality. Seven patients required intensive care, 8 required mechanical ventilation, and 7 died. Five of the patients, including 4 of the fatalities, were previously healthy.


Health workers' strikes in low-income countries: the available evidence.
To analyse the characteristics, frequency, drivers, outcomes and stakeholders of health workers' strikes in low-income countries. We reviewed the published and grey literature from online sources for the years 2009 to 2018. We used four search strategies: (i) exploration of main health and social sciences databases; (ii) use of specialized websites on human resources for health and development; (iii) customized Google search; and (iv) consultation with experts to validate findings. To analyse individual strike episodes, pre-existing conditions and influencing actors, we developed a conceptual framework from the literature. We identified 116 records reporting on 70 unique health workers' strikes in 23 low-income countries during the period, accounting for 875 days of strike. Year 2018 had the highest number of events (17), corresponding to 170 work days lost. Strikes involving more than one professional category was the frequent strike modality (32 events), followed by strikes by physicians only (22 events). The most commonly reported cause was complaints about remuneration (63 events), followed by protest against the sector's governance or policies (25 events) and safety of working conditions (10 events). Positive resolution was achieved more often when collective bargaining institutions and higher levels of government were involved in the negotiations. In low-income countries, some common features appear to exist in health sector strikes' occurrence and actors involved in such events. Future research should focus on both individual events and regional patterns, to form an evidence base for mechanisms to prevent and resolve strikes.


An ambiguous presentation of cardiac calcified amorphous tumor in a 37-year-old male.
Calcified amorphous tumors are extremely rare benign cardiac lesions that can emerge in any chamber, and have many clinical obscurities. We herein report a case of a mid-aged previously healthy male with a complaint of recurrent chest pain. Echocardiogram revealed the presence of right atrial mass that was confirmed by transesophageal echocardiography. The patient underwent total resection of the mass with attached atrial wall. Pathological studies confirmed the diagnosis of calcified amorphous tumor. Five-year follow-up was uneventful with total symptomatic relief. In conclusion, we recommend considering the diagnosis of calcified amorphous tumors in any patient with intracardiac mass.


Site-selective differences in cytochrome P450 isoform activities. Comparison of expression in rat and rhesus monkey lung and induction in rats.
The distribution of pulmonary cytochrome P450 (P450 or CYP) isoforms has been investigated primarily in immunohistochemical studies, which are neither quantitative nor reflective of the functions of these enzymes. Studies of enzyme activities have been performed using whole-lung homogenates or isolated cells, but there is little information on the regioselective expression of P450 monooxygenases. The aims of this study were to compare the activities of P450 monooxygenases in different lung subcompartments in two commonly studied animal models, i.e. rats and monkeys, and to explore the possibility that inducing agents would result in activity up-regulation that is highly site-selective, using rats as a model. Microdissection techniques were used to separate the airways from blood vessels and lung parenchyma. In rats, CYP1A1 (ethoxyresorufin) and CYP2B (pentoxyresorufin) dealkylase activities were highest in the parenchyma, whereas CYP2E1 (p-nitrophenol) hydroxylase activity was highest in the airways. P450 reductase activities were similar in airways and parenchyma and were lower in trachea. In monkeys, no significant site-selective differences in CYP1A1 and CYP2B1 activities were found. In contrast, CYP2E1 activity was higher in the distal bronchioles and parenchyma than in the proximal airways. P450 reductase activities were similar in microsomes prepared from all subcompartments of monkey lung. Induction of rat CYP1A1 activity by beta-naphthoflavone (administered ip) was much greater in the airways and lung parenchyma ( approximately 30-fold) than in the liver ( approximately 10-fold) or trachea ( approximately 2.5-fold). Oral administration of phenobarbital or acetone increased CYP2B and CYP2E1 activities in rat liver but had no significant effect on P450 activities in subcompartments of rat lung. These findings support the conclusion that there are regiospecific and species-specific differences in the activities of P450 isoforms and that the inducibility of rat pulmonary P450s is dependent on the isoform and lung region.


A study protocol of the photo-supported conversations about the well-being intervention (Be Well™) for people with stress related disorders.
Stress-related illnesses constitute a huge problem in society. The primary care services in Sweden form the first line of care whose role is to coordinate interventions for reducing symptoms, as well as health-promoting interventions. There is lack of knowledge concerning health-promoting interventions for these illnesses. The aim of this study is to evaluate whether photo-supported conversations about well-being (Be Well™) as an intervention, in addition to care as usual within the primary care services, improves health and well-being for patients with stress-related illnesses. The intervention will be compared to a control group, who receive care as usual. A further aim is to conduct a process evaluation. This ongoing project has a quasi-experimental design, using quantitative and qualitative methods, and includes patients from primary care centres in two Swedish counties. Seventy patients, 20-67 years, with stress-related illnesses will be recruited. They constitute an intervention group, which receive the intervention together with care as usual, and a control group, which receive care as usual. The intervention, photo-supported conversations about well-being, involves 12 sessions. Care as usual entails medication, occupational therapy, physiotherapy and/or psychotherapy. Data collection is carried out at baseline, and outcomes are assessed directly after the intervention, as well as six months after completion of the intervention. The outcomes are evaluated based on factors related to health, well-being and everyday occupations. Furthermore, data concerning experiences of well-being and perceptions of the intervention will be collected in interviews. The therapists will also be interviewed about their experiences of performing the intervention. Data will be analysed with non-parametric statistics, and qualitative methodology. The project is based on the concept that focusing on well-being despite living with stress-related illness may positively impact health and well-being as well as activity-related aspects, and that photo-supported conversations about well-being can contribute a complement to other treatment and rehabilitation. A strength is the use of a wide range of methods: such as quantitative measures, photographs, and qualitative interviews with participants and therapists. The results will thus provide knowledge about potential effects of this health-promoting intervention. Trial registration Clinical Trials.gov: NCT04832295; retrospectively registered 2nd April 2021 https://clinicaltrials.gov/ct2/show/NCT04832295.


Assessment of a novel scoring method based on solvent accessible surface area descriptors.
A novel scoring algorithm based on unique solvent accessible surface area (SASA) descriptors was comparatively evaluated for its database enrichment potential against the virtual screening (VS) methods GOLD and Glide. Several protein test cases, including adenosine deaminase and estrogen receptor alpha, were used for the evaluation. The structure-based VS method GOLD was used to generate the protein-ligand docking poses. These docking poses were then postprocessed with a protein-ligand interaction fingerprint metric. Next, the SASA descriptors were computed for each ligand and its respective protein in their bound/unbound states; a Bayesian model was learned with SASA descriptors and subsequently used to score the remaining ligands in the screening databases. Early database enrichments using SASA descriptors were found comparable or superior to those of GOLD and Glide. Moreover, SASA descriptors display an outstanding robustness to produce satisfactory early enrichments for a large variety of target classes. Based on these encouraging results, these novel topological descriptors constitute a valuable in silico tool in hit finding practices.


Successful management of discovered pH dependence in vancomycin recovery studies: novel HPLC method for microdialysis and plasma samples.
Vancomycin is a glycopeptide antibiotic approved for the treatment of serious infections or patients allergic to beta-lactams. A rapid HPLC assay using UV detection for the determination in microdialysate and human plasma was developed. After sample preparation, using methanol and trichloroacetic acid for plasma and water for microdialysate, 20 microL were injected and separated on a RP(18) column. Overall, the assay exhibited good precision and accuracy. The diffusion properties of vancomycin investigated in in vitro microdialysis experiments revealed an unfavourable concentration dependence avertable by keeping a constant pH using phosphate buffer as perfusate. The mean relative recoveries were 27.8% [coefficient of variation (CV) 11.1%] and 33.2% (CV 8.3%) for retrodialysis and recovery experiments, respectively. Following characterization of vancomycin in in vitro microdialysis, the developed setting is suitable for application in (pre-)clinical studies.


Altered intrinsic functional connectivity in the latent period of epileptogenesis in a temporal lobe epilepsy model.
The latent period, a seizure-free phase, is the duration between brain injury and the onset of spontaneous recurrent seizures (SRSs) during epileptogenesis. The latent period is thought to involve several progressive pathophysiological events that lead to the evolution of the chronic epilepsy phase. Hence, it is vital to investigate the changes in the latent period during epileptogenesis in order to better understand temporal lobe epilepsy (TLE), and to achieve early diagnosis and appropriate management of the condition. Accordingly, recent studies with patients with TLE using resting-state functional magnetic resonance imaging (rs-fMRI) have reported that alterations of resting-state functional connectivity (rsFC) during the chronic period are associated with some clinical manifestations, including learning and memory impairments, emotional instability, and social behavior deficits, in addition to repetitive seizure episodes. In contrast, the changes in the intrinsic rsFC during epileptogenesis, particularly during the latent period, remain unclear. In this study, we investigated the alterations in intrinsic rsFC during the latent and chronic periods in a pilocarpine-induced TLE mouse model using intrinsic optical signal imaging (IOSI). This technique can monitor the changes in the local hemoglobin concentration according to neuronal activity and can help investigate large-scale brain intrinsic networks. After seeding on the anatomical regions of interest (ROIs) and calculating the correlation coefficients between each ROI, we established and compared functional correlation matrices and functional connectivity maps during the latent and chronic periods of epilepsy. We found a decrease in the interhemispheric rsFC at the frontal and temporal regions during both the latent and chronic periods. Furthermore, a significant decrease in the interhemispheric rsFC was observed in the somatosensory area during the chronic period. Changes in network configurations during epileptogenesis were examined by graph theoretical network analysis. Interestingly, increase in the power of low frequency oscillations was observed during the latent period. These results suggest that, even if there are no apparent ictal seizure events during the latent period, there are ongoing changes in the rsFC in the epileptic brain. Furthermore, these results suggest that the pathophysiology of epilepsy may be related to widespread altered intrinsic functional connectivity. These findings can help enhance our understanding of epileptogenesis, and accordingly, changes in intrinsic functional connectivity can serve as an early diagnosis.


Axotomy induces the expression of vasopressin receptors in cranial and spinal motor nuclei in the adult rat.
8-L-Arginine vasopressin ([Arg8]VP) receptors are expressed transiently in the rat facial nucleus during the perinatal period. Electrophysiological studies suggest that at least part of these receptors is located on facial motoneurones. In the present study we report that, in the adult rat, unilateral section of a facial nerve results in a massive and transient reexpression of [Arg8]VP receptors in the deeferented facial nucleus. Data were obtained by quantitative film autoradiography. During the first 2 postoperative weeks, binding of an iodinated ligand selective for V1a-type receptors increased about 10-fold. Maximal levels of binding were maintained for 1-2 weeks and then started to decrease. Binding was not strictly restricted to the facial nucleus but included the neuropile between motoneuronal pools and the perifacial area, which may indicate a dendritic localization of [Arg8]VP receptors. To investigate whether other motor nuclei also react to axotomy by up-regulating [Arg8]VP receptors, we sectioned either a hypoglossal nerve or a sciatic nerve. Two weeks after surgery, the hypoglossal nucleus or sciatic motoneuronal pools ipsilateral to the lesion were intensely labeled with the iodinated ligand. In contrast, nerve section had no effect on oxytocin binding sites in facial, hypoglossal, or sciatic motor nuclei. The results suggest that [Arg8]VP receptor expression in motor nuclei may depend upon neuromuscular contacts and, thus, that [Arg8]VP may be involved in the establishment of neuromuscular connections during development and in their reestablishment after nerve injury.


Effect of iron on cholesterol 7α-hydroxylase expression in alcohol-induced hepatic steatosis in mice.
Both iron and lipids are involved in the progression of alcoholic fatty liver disease (AFLD), but the interaction between iron and lipids in AFLD is unclear. Here, we tested the hypothesis that iron regulates the expression of genes involved in lipid metabolism through iron regulatory proteins (IRPs), which interact with the iron-responsive elements (IREs) in the untranslated regions (UTRs) of genes, resulting in lipid accumulation. Using "RNA structure software", we predicted the mRNA secondary structures of more than 100 genes involved in lipid metabolism to investigate whether the IRE structure exists in novel mRNAs. Cholesterol 7α-hydroxylase (Cyp7a1) has an IRE-like stem-loop, a noncanonical IRE structure, in its 3'-UTR. Cyp7a1 expression can be regulated by in vivo and in vitro iron treatment. In addition, the noncanonical IRE motif can efficiently bind both to IRP1 and IRP2. The results indicate that hepatic iron overloading in AFLD mice decreased Cyp7a1 expression and resulted in cholesterol accumulation, providing a new mechanism of iron-regulated gene transcription and translation through the interaction between iron and a noncanonical IRE structure in Cyp7a1 mRNA. This finding has significant implications in studying a proposed mechanism for the regulation of cholesterol homeostasis by an Fe/IRP/noncanonical IRE axis.


Opinion disparity in hypergraphs with community structure.
The division of a social group into subgroups with opposing opinions, which we refer to as opinion disparity, is a prevalent phenomenon in society. This phenomenon has been modeled by including mechanisms such as opinion homophily, bounded confidence interactions, and social reinforcement mechanisms. In this paper, we study a complementary mechanism for the formation of opinion disparity based on higher-order interactions, i.e., simultaneous interactions between multiple agents. We present an extension of the planted partition model for uniform hypergraphs as a simple model of community structure, and we consider the hypergraph Susceptible-Infected-Susceptible (SIS) model on a hypergraph with two communities where the binary ideology can spread via links (pairwise interactions) and triangles (three-way interactions). We approximate this contagion process with a mean-field model and find that for strong enough community structure, the two communities can hold very different average opinions. We determine the regimes of structural and infectious parameters for which this opinion disparity can exist, and we find that the existence of these disparities is much more sensitive to the triangle community structure than to the link community structure. We show that the existence and type of opinion disparities are extremely sensitive to differences in the sizes of the two communities.


Health information for young people where and when they most want it: a case study of www.teenagehealthfreak.org.
The Internet is an exciting resource for providing immediately available, evidence-based, health information for young people in an age-appropriate form on a 24 hours/day, 7 days/week basis. www.teenagehealthfreak.org is a United Kingdom-based Web site designed to take advantage of this. The content of the site, which is the leading teenage health Web site on a Google search, contains both the diary of a hypochondriac 15-year-old boy and a virtual doctor's surgery. It also allows for young people to e-mail health-related questions and receive relevant answers from a health expert. Analysis of the content of these e-mails indicates the unmet health needs and concerns of young people. Future developments of the site include linking the site www.youthhealthtalk.org, a Web site that contains videotaped interviews with young people who have a variety other health concerns.


Boosting Electrocatalytic Nitrate-to-Ammonia via Tuning of N-Intermediate Adsorption on a Zn-Cu Catalyst.
The renewable-energy-powered electroreduction of nitrate (NO3 - ) to ammonia (NH3 ) has garnered significant interest as an eco-friendly and promising substitute for the Haber-Bosch process. However, the sluggish kinetics hinders its application at a large scale. Herein, we first calculated the N-containing species (*NO3 and *NO2 ) binding energy and the free energy of the hydrogen evolution reaction over Cu with different metal dopants, and it was shown that Zn was a promising candidate. Based on the theoretical study, we designed and synthesized Zn-doped Cu nanosheets, and the as-prepared catalysts demonstrated excellent performance in NO3 - -to-NH3 . The maximum Faradaic efficiency (FE) of NH3 could reach 98.4 % with an outstanding yield rate of 5.8 mol g-1 h-1 , which is among the best results up to date. The catalyst also had excellent cycling stability. Meanwhile, it also presented a FE exceeding 90 % across a wide potential range and NO3 - concentration range. Detailed experimental and theoretical studies revealed that the Zn doping could modulate intermediates adsorption strength, enhance NO2 - conversion, change the *NO adsorption configuration to a bridge adsorption, and decrease the energy barrier, leading to the excellent catalytic performance for NO3 - -to-NH3 .


Lack of clinical significance of the ImmuKnow(TM)-Cylex assay for the detection of cellular immune function in patients with renal cell carcinoma.
This study aimed to explore the clinical value of the CD4(+) T cell ATP levels in patients with renal cell carcinoma through the application of the ImmuKnow(TM)-Cylex(®) assay. We recruited 104 patients with renal cancer who had undergone surgery at Fuzhou General Hospital from March 2009 to June 2012, and were subsequently treated by dendritic cell and cytokine-induced killer cell bio-therapy or interferon-α therapy. The changes in CD4(+) T cell ATP levels were detected at the perioperative period and at 10 days, 1 month, 3 months, and 1 year after the surgery using the ImmuKnow assay. In addition, the differences in ATP levels in different therapy groups were compared and the prognosis conditions were analyzed. Our results demonstrated that no significant difference in the ATP levels occurred at different time points; furthermore, there were no obviously different ATP levels between the different therapy groups, and the ATP levels were found to have no clinical significance for the assessment of renal cancer prognosis. Overall, this study suggested that CD4(+) T cell ATP levels as detected by the ImmuKnow assay have no obvious clinical value in patients with renal cancer.


Reversible Regulation of Polyubiquitin Gene UBC via Modified Inducible CRISPR/Cas9 System.
Ubiquitin is required under both normal and stress conditions. Under stress conditions, upregulation of the polyubiquitin gene UBC is essential to meet the requirement of increased ubiquitin levels to confer stress resistance. However, UBC upregulation is usually observed only under stress conditions and not under normal conditions. Therefore, it has not been possible to upregulate UBC under normal conditions to study the effect of excess ubiquitin on cellular machinery. Recently, the CRISPR/Cas9 system has been widely used in biological research as a useful tool to study gene disruption effects. In this study, using an inducible CRISPR/Cas9 variant, a dCas9-VP64 fusion protein, combined with a single guide RNA (sgRNA) containing MS2 aptamer loops and MS2-p65-HSF1, we developed a system to increase the ubiquitin pool via upregulation of UBC. Although it is challenging to upregulate the expression of a gene that is already expressed at high levels, the significance of our system is that UBC upregulation can be induced in an efficient, reversible manner that is compatible with cellular processes, even under normal conditions. This system can be used to study ubiquitin pool dynamics and it will be a useful tool in identifying the role of ubiquitin under normal and stress conditions.


Exploiting multicompartment effects in triple-echo steady-state T2 mapping for fat fraction quantification.
To investigate and exploit the effect of intravoxel off-resonance compartments in the triple-echo steady-state (TESS) sequence without fat suppression for T2 mapping and to leverage the results for fat fraction quantification. In multicompartment tissue, where at least one compartment is excited off-resonance, the total signal exhibits periodic modulations as a function of echo time (TE). Simulated multicompartment TESS signals were synthesized at various TEs. Fat emulsion phantoms were prepared and scanned at the same TE combinations using TESS. In vivo knee data were obtained with TESS to validate the simulations. The multicompartment effect was exploited for fat fraction quantification in the stomach by acquiring TESS signals at two TE combinations. Simulated and measured multicompartment signal intensities were in good agreement. Multicompartment effects caused erroneous T2 offsets, even at low water-fat ratios. The choice of TE caused T2 variations of as much as 28% in cartilage. The feasibility of fat fraction quantification to monitor the decrease of fat content in the stomach during digestion is demonstrated. Intravoxel off-resonance compartments are a confounding factor for T2 quantification using TESS, causing errors that are dependent on the TE. At the same time, off-resonance effects may allow for efficient fat fraction mapping using steady-state imaging. Magn Reson Med 79:423-429, 2018. © 2017 International Society for Magnetic Resonance in Medicine.


The formative years: medical ethics comes of age.
When the Journal of Medical Ethics first appeared in April 1975, the prospects of success seemed uncertain. There were no scholars specialising in the field, the readership could not be guaranteed, and the medical profession itself seemed, at the very least, ambivalent about a subject thought by many to be the province of doctors alone, to be acquired through an apprenticeship model, and certainly not taught or examined in any formal sense. However, change was afoot, fresh scandals created an awareness that outside help was needed to think through the new challenges facing the profession, and the success of the medical groups revealed a clear way forward through multidisciplinary and critically reflective discussion of the host of emerging ethical and legal issues. In this article the formative years of the journal are recaptured, with a claim that the core principles on which it was founded must endure if it is to continue to 'do good medical ethics' over the next 40 years.


Polymerase chain reaction based detection of bacterial 16S rRNA gene in the cerebrospinal fluid in the diagnosis of bacterial central nervous system infection in the course of external cerebrospinal fluid drainage. Comparison with standard diagnostics currently used in clinical practice.
External drainage of cerebrospinal fluid (CSF) is a commonly used neurosurgical procedure. Complications of the procedure comprise central nervous system (CNS) bacterial infections, the frequency of which is estimated at around 6-10%. Detection of these infections is ineffective in many cases. The aim of the study was to evaluate the usefulness of a polymerase chain reaction (PCR)-based detection of bacterial 16S rRNA gene (16S rDNA) in the CSF. The study group consisted of 50 patients. Clinical signs of CNS infection were monitored and routine laboratory and microbiological tests were performed. The results of standard methods were compared with the bacterial 16S rDNA detection. Using cultures, CNS infection was diagnosed in 8 patients, colonization of the drainage catheter in 6 patients, and sample contamination in 7 patients. In the group of the remaining 29 patients, no positive CSF culture was obtained and 13 of these patients also had all negative results for 16S rDNA detection. For the remaining 16 patients of this group, CNS infection, colonization of the catheter and sample contamination were diagnosed via PCR alone. Routine biochemical CSF tests and blood inflammatory parameters had a supporting value. Routine hospital tests do not provide rapid and efficient detection of the external drainage related bacterial CNS infection. It is justified to use several diagnostic methods simultaneously. The16S rDNA determination in CSF can increase the probability of detection of possible pathogens.


Cisplatin-Induced Renal Salt Wasting Requiring over 12 Liters of 3% Saline Replacement.
Cisplatin is known to induce Fanconi syndrome and renal salt wasting (RSW). RSW typically only requires transient normal saline (NS) support. We report a severe RSW case that required 12 liters of 3% saline. A 57-year-old woman with limited stage small cell cancer was admitted for cisplatin (80 mg/m2) and etoposide (100 mg/m2) therapy. Patient's serum sodium (SNa) decreased from 138 to 133 and 125 mEq/L within 24 and 48 hours of cisplatin therapy, respectively. A diagnosis of syndrome of inappropriate antidiuretic hormone secretion (SIADH) was initially made. Despite free water restriction, patient's SNa continued to decrease in association with acute onset of headaches, nausea, and dizziness. Three percent saline (3%S) infusion with rates up to 1400 mL/day was required to correct and maintain SNa at 135 mEq/L. Studies to evaluate Fanconi syndrome revealed hypophosphatemia and glucosuria in the absence of serum hyperglycemia. The natriuresis slowed down by 2.5 weeks, but 3%S support was continued for a total volume of 12 liters over 3.5 weeks. Attempts of questionable benefits to slow down glomerular filtration included the administration of ibuprofen and benazepril. To our knowledge, this is the most severe case of RSW ever reported with cisplatin.


Side effects of mandibular advancement splints for the treatment of snoring and obstructive sleep apnea: a systematic review.
Occlusal side effects or development of pain and/or functional impairment of the temporomandibular complex are potential reasons for poor compliance or abandonment of mandibular advancement splints treatment for snoring and obstructive sleep apnea. This study aimed at providing a comprehensive review evaluating the craniofacial side effects of oral appliance therapy for snoring and obstructive sleep apnea. An electronic search was systematically conducted in PubMed and Virtual Health Library from their inception until October 2016. Only Randomized Controlled Trials whose primary aim was to measure objectively identified side effects on craniofacial complex of a custom-made oral appliance for treating primary snoring or obstructive sleep apnea were included. Studied patients should be aged 20 or older. The risk of bias in the trials was assessed in accordance with the recommendations of The Cochrane Risk of Bias criteria. A total of 62 full-text articles were assessed for eligibility. After the review process, only 6 met all the inclusion criteria. All studies were rated as having a high risk of bias. The most uniformly reported mandibular advancement splint side effects were predominantly of dental nature and included a decrease in overjet and overbite. The risk of developing pain and function impairment of the temporomandibular complex appeared limited with long-term mandibular advancement splint use. The limited available evidence suggests that mandibular advancement splint therapy for snoring and obstructive sleep apnea results in changes in craniofacial morphology that are predominantly dental in nature, specially on a long-term basis. Considering the chronic nature of obstructive sleep apnea and that oral appliance use might be a lifelong treatment, a thorough customized follow-up should therefore be undertaken to detect possible side effects on craniofacial complex. It is also important to provide adequate information to the patients regarding these possible changes, especially to those in whom larger occlusal changes are to be expected or in whom they are unfavorable. Long-term assessments of adverse effects of oral appliance therapy, with larger study samples and recruitment of homogenous patient population are still required.


Left ventricle changes early after breath-holding in deep water in elite apnea divers.
To study by ultrasounds cardiac morphology and function early after breath-hold diving in deep water in elite athletes. Fifteen healthy male divers (age 28 +/- 3 years) were studied using Doppler-echocardiography, immediately before (basal condition, BC) and two minutes after breath-hold diving (40 meters, acute post-apnea condition, APAC). Each subject performed a series of three consecutive breath-hold dives (20-30 and 40 m depth). End-diastolic left ventricular (LV) diameter (EDD) and end-diastolic LV volume (EDV) increased significantly (p < 0.01). Stroke volume (SV), cardiac index (CI), septal and posterior systolic wall-thickening (SWT) also significantly increased after diving (p < 0.01). No wall motion abnormalities were detected, and wall motion score index was unchanged between BC and APAC. Doppler mitral E wave increased significantly (p < 0.01), whereas the A wave was unchanged. Systemic vascular resistance (SVR) decreased significantly after diving (p < 0.05). In the factor analysis, filtering out the absolute values smaller than 0.7 in the loading matrix, it resulted that factor I consists of EDV, posterior SWT, SV and CI, factor II of diastolic blood pressure, waves A and E and factor III of heart rate and SVR. Systo-diastolic functions were improved in the early period after deep breath-hold diving due to favorable changes in loading conditions relative to pre-diving, namely the recruitment of left ventricular preload reserve and the reduction in afterload.


Efficient transformation of rice (Oryza sativa L.) mediated by Agrobacterium and sequence analysis of the boundaries of the T-DNA.
A large number of morphologically normal, fertile, transgenic rice plants were obtained by co-cultivation of rice tissues with Agrobacterium tumefaciens. The efficiency of transformation was similar to that obtained by the methods used routinely for transformation of dicotyledons with the bacterium. Stable integration, expression and inheritance of transgenes were demonstrated by molecular and genetic analysis of transformants in the R0, R1 and R2 generations. Sequence analysis revealed that the boundaries of the T-DNA in transgenic rice plants were essentially identical to those in transgenic dicotyledons. Calli induced from scutella were very good starting materials. A strain of A. tumefaciens that carried a so-called 'super-binary' vector gave especially high frequencies of transformation of various cultivars of japonica rice that included Koshihikari, which normally shows poor responses in tissue culture.


Predictors of spontaneous remission and recovery among women with untreated perinatal depression in India and Pakistan.
Mothers with perinatal depression can show different symptom trajectories and may spontaneously remit from depression, however, the latter is poorly understood. This is the first study which sought to investigate predictors of spontaneous remission and longer-term recovery among untreated women with perinatal depression. We analysed data from two randomised controlled trials in women with perinatal depression in India and Pakistan. Analyses were restricted to women in the control groups who did not receive active treatment. Generalised estimating equations and logistic regressions were used to estimate odds ratios (ORs) and 95% confidence intervals (CIs), adjusting for within-person correlation. In multivariable analyses, remission was associated with a husband who is not working (adjusted OR, aOR = 2.04, 95% CI 1.02-4.11), lower Patient Health Questionnaire-9 score at baseline (aOR = 0.43, 95% CI 0.20-0.90 for score of ≥20 vs. 10-14) and better social support at baseline (aOR = 2.37, 95% CI 1.32-4.27 for high vs. low social support). Women with low baseline severity may remit from perinatal depression with adequate social support from family and friends. These factors are important contributors to the management of perinatal depression and the prevention of clinical worsening, and should be considered when designing low-threshold psychological interventions.


Postoperative pain control by single doses of piroxicam administered sublingually and aspirin.
The objective of this study is to evaluate and compare the analgesic efficacy of Piroxicam Fast Dissolving Dosage Formulation (FDDF) administered sublingually either preoperatively or postoperatively with that of aspirin and placebo. Hundred patients, undergoing surgical removal of mandibular third molars were given sublingually either piroxicam FDDF 40 mg or placebo or aspirin 500 mg according to a double blind experimental model. Pain scores and pain relief was recorded hourly for six hours following the operation. Rescue analgesics used during the trial and overall assessment of the drug were recorded at the end of six hours. Scores were evaluated statistically by student t test. Piroxicam FDDF was significantly (p < 0.05) more effective than aspirin and placebo for all variables, and less rescue analgesics were required in patients receiving piroxicam preoperatively. It was concluded that single doses of piroxicam FDDF administered sublingually either preoperatively or postoperatively appeared to be effective analgesic for the control of pain after third molar surgery with no side effects.


Spectral Narrowing Accompanies Enhanced Spatial Resolution in Saturated Coherent Anti-Stokes Raman Scattering (CARS): Comparisons of Experiment and Theory.
We demonstrate theoretically and confirm experimentally the mechanism by which spectral narrowing accompanies enhanced spatial resolution in a saturated coherent anti-Stokes Raman scattering (CARS) signal that is demodulated at the third harmonic (3f) of the pump modulation frequency (f). Under these modulation conditions, theory predicts a narrowing of the full width at half-maximum (FWHM) of the CARS spectrum by a factor of 2.0 with respect to that of the spectrum obtained by demodulation at the fundamental frequency. Theory also predicts an improvement of spatial resolution by a factor of 1.7. Experimentally, narrowing of the FWHM of the CARS spectrum of 1,4-bis((E)-2-methylstyryl) benzene (MSB) crystals by a factor of 2.5 is observed upon saturation. Further experimental confirmation is provided from investigating diamond particles, for which spectral narrowing was enhanced by a factor of 2.8 and spatial resolution was enhanced by a factor of 2. Details of the mechanism and execution of the saturated CARS experiment are elucidated and limits to its applicability are suggested, one of which is the conclusion that the saturation approach is not suitable for extraction of harmonics beyond 3f. In this work, we have developed a more comprehensive understanding of the correlation between the observed experimental results and experimental factors than has been previously reported.


DPC-423 Bristol-Myers Squibb.
DPC-423 is a biphenylamine-containing amide which is under development by Bristol-Myers Squibb (formerly DuPont Pharmaceuticals) as a Factor Xa inhibitor for the potential treatment of thrombotic disorders [372092]. As of August 2000, DPC-423 was in phase I trials [380880]. DPC-423 was discovered as a result of SAR modifications of DuPont's SN-429, including replacement of the benzamidine moiety with a less basic benzylamine [398701]. Its 2-aminomethylphenyl analog DPC-602 is also under investigation for thrombotic disorders [402714].


Antibacterial activities and antioxidant capacity of Aloe vera.
The aim of this study was to identify, quantify, and compare the phytochemical contents, antioxidant capacities, and antibacterial activities of Aloe vera lyophilized leaf gel (LGE) and 95% ethanol leaf gel extracts (ELGE) using GC-MS and spectrophotometric methods. Analytically, 95% ethanol is less effective than ethyl acetate/diethyl ether or hexane (in the case of fatty acids) extractions in separating phytochemicals for characterization purposes. However, although fewer compounds are extracted in the ELGE, they are approximately 345 times more concentrated as compared to the LGE, hence justifying ELGE use in biological efficacy studies in vivo. Individual phytochemicals identified included various phenolic acids/polyphenols, phytosterols, fatty acids, indoles, alkanes, pyrimidines, alkaloids, organic acids, aldehydes, dicarboxylic acids, ketones, and alcohols. Due to the presence of the antioxidant polyphenols, indoles, and alkaloids, the A. vera leaf gel shows antioxidant capacity as confirmed by ORAC and FRAP analyses. Both analytical methods used show the non-flavonoid polyphenols to contribute to the majority of the total polyphenol content. Three different solvents such as aqueous, ethanol, and acetone were used to extract the bioactive compounds from the leaves of A. vera to screen the antibacterial activity selected human clinical pathogens by agar diffusion method. The maximum antibacterial activities were observed in acetone extracts (12 ± 0.45, 20 ± 0.35, 20 ± 0.57, and 15 ± 0.38 nm) other than aqueous and ethanol extracts. Due to its phytochemical composition, A. vera leaf gel may show promise in alleviating symptoms associated with/or prevention of cardiovascular diseases, cancer, neurodegeneration, and diabetes.


Greater than the Sum: Applying Daily-Dose Equivalents to Antipsychotic Prescription Claims to Study Real-World Effects.
Traditional methods to standardize exposures in pharmacoepidemiologic studies, like defined daily-doses, may be inadequate to capture drug class effects when there are many in-class medications, formulations, and administration routes. Antipsychotic medications are one example of a drug class with these complexities. Direct dose conversion methods are pharmacologically-based but often overlooked, potentially for lack of real-world guidance and examples of their implementation. The purpose of this article is to describe a method to implement dose conversion, using an example study that quantifies antipsychotic use among a cohort of older adults with dementia. We identified 45,442 older adults (aged ≥66 years) with dementia initiating antipsychotic therapy between January 1, 2009 and December 31, 2012 in Ontario, Canada using linked administrative healthcare databases. We developed and applied a data cleaning and dose conversion algorithm to quantify antipsychotic exposure in chlorpromazine dose equivalents at initiation, month 6, and month 12 of therapy. Results were stratified by route of administration. At initiation, 14% of patients received multiple antipsychotic prescriptions simultaneously. Patients initiating regular injectable and multiple administration routes received the highest median chlorpromazine equivalent daily-doses. Data cleaning changed 3, 16, 36, and 42% of total equivalent daily-doses in patients initiating oral, regular injectable, long-acting injectable, and multiple administration routes, respectively. Dose conversion of prescription claims data was a feasible method to quantify and present antipsychotic drug exposures. Dose conversion methods can be considered for drug effects studies of antipsychotic therapies and other medication classes with complex use.


Insights into the emerging cardiometabolic prevention and management of diabetes mellitus.
Cardiovascular disease (CVD) and Type 2 diabetes mellitus (DM2), once conceived as different entities, share common origins and pathways. Increased activity of the renin-angiotensin-aldosterone-system, insulin resistance, chronic low-grade inflammation and oxidative stress collectively contribute to endothelial dysfunction and atherosclerosis, which manifest clinically as CVD. Nowadays, it is possible to identify and intervene in high-risk populations even before the clinical diagnosis of DM2. The control of dietary patterns and increased physical activity is completely feasible, as well as the management of hypertension and dyslipidaemia. Pharmacological interventions targeted at blocking renin-angiotensin-aldosterone-system and sensitising to insulin have a role in the prevention of DM2 and CVD, and are avidly explored worldwide. In the near future, ongoing trials should provide data that will allow us to better treat patients with the cardiometabolic syndrome and diabetes in order to reduce CVD morbidity and mortality.


[Activity and distribution of plasma platelet-activating factor acetylhydrolase in polycystic ovary syndrome patients with insulin resistance].
To investigate the relationship between activity of plasma platelet-activating factor acetylhydrolase (PAF-AH) and insulin resistance (IR) in patients with polycystic ovary syndrome (PCOS). From Oct. 2006 to Jan. 2008, 105 PCOS patients undergoing treatment in Department of Obstetrics and Gynecology of Second Hospital affiliated to West China University were enrolled in the study, among 53 cases with homeostatic model assessment IR (HOMA-IR) exceed or equal 2.77 as IR-PCOS group and 52 cases with HOMA-IR less than 2.77 as non IR-PCOS group. In the mean time, 53 infertile women due to fallopian tube or husband factors were chosen as control group. Plasma PAF-AH activity, high-density lipoprotein-associated PAF-AH (H-PAF-AH) activity, low-density lipoprotein-associated PAF-AH (L-PAF-AH) activity, the ratio of L-PAF-AH to H-PAF-AH activity were measured and compared among three groups. The method of Pearson correlation analysis and stepwise multiple regression analysis were used to study the relationship between the ratio of L-PAF-AH to H-PAF-AH activities and hormonal and metabolic parameters. (1) Plasma PAF-AH activity [(0.055 ± 0.012) mmol x L(-1) x min(-1)], L-PAF-AH activity [(0.052 ± 0.012) mmol x L(-1) x min(-1)], and the ratio of L-PAF-AH to H-PAF-AH activity (23 ± 6) in the IR-PCOS group were significantly higher than those at control group [(0.050 ± 0.009) mmol x L(-1) x min(-1), (0.047 ± 0.009) mmol x L(-1) x min(-1) and (18 ± 4)] and non-IR-PCOS group [(0.050 ± 0.0012) mmol x L(-1) x min(-1), (0.048 ± 0.012) mmol x L(-1) x min(-1) and (18 ± 5), P < 0.05]. The ratio of L-PAF-AH to H-PAF-AH activities at IR-PCOS group was also significantly higher than those at Non IR-PCOS and control group after correction for body mass index (P < 0.01). (2) Pearson correlation analysis showed that the ratio of L-PAF-AH to H-PAF-AH in PCOS patients was positively correlated with atherogenic index, age, body mass index, waist-to-hip ratio, HOMA-IR, triglyceride levels, fasting insulin levels, fasting glucose levels, low density lipoprotein cholesterol and total cholesterol (r = 0.644, 0.247, 0.296, 0.212, 0.356, 0.587, 0.377, 0.375, 0.292 and 0.199, respectively, P < 0.05), and negatively correlated with high density lipoprotein cholesterol (r = -0.510, P < 0.05). The multiple stepwise regression analysis showed that atherogenic index and triglyceride were significant variables for the ratio of L-PAF-AH to H-PAF-AH activity (standardized coefficient, beta = 0.515 and 0.201, respectively, P < 0.05). The elevated L-PAF-AH activity may be associated with IR in PCOS patients, and the increased ratio of L-PAF-AH to H-PAF-AH activities may be a potential marker of inflammation in the patients.


Iron deficiency and hemolytic anemia reversed by ventricular septal myectomy.
Hemolytic anemia has been reported to occur in the setting of aortic stenosis and prosthetic heart valves, but much more rarely in association with obstructive hypertrophic cardiomyopathy (HC). Of the few descriptions of hemolytic anemia secondary to HC, all but one case involved bacterial endocarditis contributing to left ventricular outflow tract obstruction. We present the case of a 67-year-old man with recurrent hemolytic anemia and HC, without infective endocarditis. Attempts at iron repletion and augmentation of beta-blocker therapy proved his anemia to be refractory to medical management. Ventricular septal myectomy led to the resolution of hemolysis, anemia, and its coexisting symptoms.


The effect of the addition of carbon nanotubes in the hydrothermal synthesis and in the thermal phase stability of nanozirconia.
The yttria partially stabilized zirconia is a very attractive material for orthopaedic applications. It exhibits excellent biocompatibility, high fracture toughness, high strength and low wear rates. But case studies show that delayed failure can occur in vivo due to crack propagation. Carbon nanotubes could avoid the slow crack propagation and enhance the toughness of the ceramic material used for prostheses fabrication. In this work, X-ray diffraction has been used to study the influence of the addition of MWCNT on the hydrothermal synthesis of tetragonal zirconia nanoparticles and on the phase stability of the CNT-nanozirconia nanocomposite with the temperature. First, the influence of the processing variables on the hydrothermal synthesis has been studied. The theoretical mathematical models that relate the percentage of tetragonal zirconia nanocrystals and the relative crystallinity with the processing variables in the range of analyzed values have been obtained. The values that give the maximum percentage and crystallinity of tetragonal phase in the studied range have been established. No significant differences were observed in the crystalline phases obtained when adding MWCNT during the synthesis. Nanozirconia partially coated MWCNT synthesized under the optimized parameters were added to commercially available nanozirconia particles and their influence in the phase stability of the zirconia with the temperature was studied by XRD. It was concluded that the addition of the carbon nanotubes delays both the monoclinic phase decomposition and the grain growth.


Generation and release of nitrotyrosine O-sulfate by HepG2 human hepatoma cells upon SIN-1 stimulation: identification of SULT1A3 as the enzyme responsible.
In addition to serving as a biomarker of oxidative/nitrative stress, elevated levels of nitrotyrosine have been shown to cause DNA damage or trigger apoptosis. Whether the body is equipped with mechanisms for protecting against the potentially harmful nitrotyrosine remains unknown. The present study was designed to investigate the possibility that sulfation serves as a pathway for the metabolism/regulation of nitrotyrosine. Using metabolic labelling, nitrotyrosine O-[35S]sulfate was found to be produced and released into the medium of HepG2 human hepatoma cells labelled with [35S]sulfate in the presence of nitrotyrosine. To identify the enzyme(s) responsible for nitrotyrosine sulfation, a systematic study of all eleven known human cytosolic SULTs (sulfotransferases) was performed. Of the 11 enzymes tested, only SULT1A3 displayed sulfating activity toward nitrotyrosine. The pH-dependence and kinetic constants of SULT1A3 with nitrotyrosine or dopamine as substrate were determined. To examine whether the sulfation of nitrotyrosine occurs in the context of cellular physiology, HepG2 cells labelled with [35S]sulfate were treated with SIN-1 (morpholinosydnonimine), a peroxynitrite generator. Increments of nitrotyrosine O-[35S]sulfate were detected in the medium of HepG2 cells treated with higher concentrations of SIN-1. To gain insight into the physiological relevance of nitrotyrosine sulfation, a time-course study was performed using [3H]tyrosine-labelled HepG2 cells treated with SIN-1. The findings confirm that the bulk of free [3H]nitrotyrosine inside the cells was present in the unconjugated form. The proportion of sulfated [3H]nitrotyrosine increased dramatically in the medium over time, implying that sulfation may play a significant role in the metabolism of free nitrotyrosine.


Costing the nursing service of patient education.
Patient Classification Systems (PCS) are one method of determining nursing service cost of patient education. A teaching classification study compares the value that different PCSs place on this aspect of nursing care. Through this comparison, information obtained helps nurses objectively update existing patient classification systems and provides guidelines for developing a system to include patient education.


Cerebrospinal fluid proteome profile in multiple sclerosis.
Cerebrospinal fluid (CSF) proteins may provide important information about the pathomechanisms present in multiple sclerosis (MS). Although diagnostic criteria for early MS are available, there is still a need for biomarkers, predicting disease subtype and progression to improve individually tailored treatment. Using the two-dimensional difference gel electrophoresis (2-D-DIGE) technology for comparative analysis, we compared CSF samples from patients with MS of the relapse-remitting type (RRMS, n = 12) and from patients with clinically isolated syndrome (CIS, n = 12) suggestive of a first demyelinating attack with neurologically normal controls. Protein spots that showed more than two-fold difference between patients and controls were selected for further analysis with MALDI-TOF mass spectrometry. Immunoblot analysis was performed to confirm the validity of individual candidate proteins. In RRMS, we identified 1 up-regulated and 10 down-regulated proteins. In CIS, 2 up-regulated and 11 down-regulated proteins were identified. One of these proteins (Apolipoprotein A1) was confirmed by immunoblot. Though the pathophysiological role of these proteins still remains to be elucidated in detail and further validation is needed, these findings may have a relevant impact on the identification of disease-specific markers.


Anhydrous octyl-glucoside phase transition from lamellar to isotropic induced by electric and magnetic fields.
A static deuterium nuclear magnetic resonance (2HNMR) technique (magnetic field, B = 7.05 T) was employed to monitor the thermotropic lamellar phase of the anhydrous 1:1 mixture sample of octyl-b-D-glucoside (βOG) and that of partially deuterium labelled at the alpha position on the chain, i.e.,βOG-d2 In the absence of an electric field, the 2H NMR spectrum of the mixture gives a typical quadrupolar doublet representing the aligned lamellar phase. Upon heating to beyond the clearing temperature at 112 °C, this splitting converts to a single line expected for an isotropic phase. Simultaneous application of magnetic and electric fields (E = 0.4 MV/m) at 85 °C in the lamellar phase, whose direction was set to be parallel or perpendicular to the magnetic field, resulted in the change of the doublet into a single line and this recovers to the initial doublet with time for both experimental geometries. This implies E- and B-field-induced phase transitions from the lamellar to an isotropic phase and a recovery to the lamellar phase again with time. Moreover, these phase transformations are accompanied by a transient current. A similar observation was made in a computational study when an electric field was applied to a water cluster system. Increasing the field strength distorts the water cluster and weakens its hydrogen bonds leading to a structural breakdown beyond a threshold field-strength. Therefore, we suggest the observed field-induced transition is likely due to a structure change of the βOG lamellar assembly caused by the field effect and not due to Joule heating.


Towards the total synthesis of chondrochloren A: synthesis of the (Z)-enamide fragment.
The stereoselective synthesis of the (Z)-enamide fragment of chondrochloren (1) is described. A Buchwald-type coupling between amide 3 and (Z)-bromide 4 was used to generate the required fragment. The employed amide 3 comprising three chiral centers was obtained through a seven-step sequence starting from ᴅ-ribonic acid-1,4-lactone. The (Z)-vinyl bromide 4 is accessible in four steps from 4-hydroxybenzaldehyde. The pivotal cross coupling between both fragments was achieved after extensive experimentation with copper(I) iodide, K2CO3 and N,N'-dimethylethane-1,2-diamine.


ProSAT+: visualizing sequence annotations on 3D structure.
PRO: tein S: tructure A: nnotation T: ool-plus (ProSAT(+)) is a new web server for mapping protein sequence annotations onto a protein structure and visualizing them simultaneously with the structure. ProSAT(+) incorporates many of the features of the preceding ProSAT and ProSAT2 tools but also provides new options for the visualization and sharing of protein annotations. Data are extracted from the UniProt KnowledgeBase, the RCSB PDB and the PDBe SIFTS resource, and visualization is performed using JSmol. User-defined sequence annotations can be added directly to the URL, thus enabling visualization and easy data sharing. ProSAT(+) is available at http://prosat.h-its.org.


Dynamic regulation of lysine acetylation: the balance between acetyltransferase and deacetylase activities.
Reversible posttranslational modification of proteins is a critically important process in physiological regulation in all tissues, including the kidney. Lysine acetylation occurs in all organisms, including prokaryotes, and is regulated by a balance between the lysine acetyltransferases (adding an acetyl group to the ε-amino group of a lysine) and deacetylases (removing it). The kidney is an organ rich with acetylated lysines, which map to >2,000 unique histone and nonhistone proteins. However, the functional significance of these modifications remains to be discovered. Here, we have compiled gene lists of the acetyltransferases and deacetylases in the mammalian genomes and mapped their mRNA expression along the renal tubule. These lists will be useful for generating targeted approaches to test the physiological or pathophysiological significance of lysine acetylation changes in the kidney.


Virtual View Generation Based on 3D-Dense-Attentive GAN Networks.
A binocular vision system is a common perception component of an intelligent vehicle. Benefiting from the biomimetic structure, the system is simple and effective. Which are extremely snesitive on external factors, especially missing vision signals. In this paper, a virtual view-generation algorithm based on generative adversarial networks (GAN) is proposed to enhance the robustness of binocular vision systems. The proposed model consists of two parts: generative network and discriminator network. To improve the quality of a virtual view, a generative network structure based on 3D convolutional neural networks (3D-CNN) and attentive mechanisms is introduced to extract the time-series features from image sequences. To avoid gradient vanish during training, the dense block structure is utilized to improve the discriminator network. Meanwhile, three kinds of image features, including image edge, depth map and optical flow are extracted to constrain the supervised training of model. The final results on KITTI and Cityscapes datasets demonstrate that our algorithm outperforms conventional methods, and the missing vision signal can be replaced by a generated virtual view.


Post-traumatic stress disorder, depression, anxiety and quality of life in patients with traffic-related injuries.
This paper reports a study to investigate and follow-up relationships between post-traumatic stress disorder, anxiety, depression and quality of life in patients after traffic-related injuries. Worldwide, traffic accidents kill 1.2 million people and injure 50 million people per year. Accidental injuries are fourth in the top five causes of death in Taiwan. For survivors, traffic accidents not only cause physical impairments, but also psychological trauma, such as post-traumatic stress disorder, depression and anxiety, all of which affect the quality of life. An exploratory, correlational design was used, and participants were recruited consecutively. Data were collected at 1 and 6 weeks post-injury for 64 patients from two major medical centres in Taiwan. Instruments were the New Injury Severity Scale, Post-traumatic Stress Disorder Reaction Index, Beck Depression Inventory, State Anxiety Inventory and Medical Outcomes Study Questionnaire. The data were collected in 2002. Statistically significant improvements occurred in depression, anxiety and the quality of life between week 1 and week 6 (P<0.05); high levels of post-traumatic stress disorder symptoms at week 1 (87.5%) and at week 6 (82.8%) showed no statistically significant improvement. There was a positive correlation between post-traumatic stress disorder and depression (r=0.70, P<0.001) and between post-traumatic stress disorder and anxiety (r=0.57, P<0.001), and a negative correlation between post-traumatic stress disorder and quality of life (r=-0.47, P<0.001). Depression was the most important variable to predict post-traumatic stress disorder at week 6, with depression levels at week 6 being a more powerful predictor than those at week 1. Regression analysis revealed that depression (19%) at week 1, depression at week 6 (45%), anxiety (3.8%) at week 6 and post-traumatic stress disorder (5.8%) explained a statistically significant amount of the variance at week 6. The findings suggest that traffic accidents have an impact on people's psychosocial wellbeing. Healthcare professionals need to implement interventions to decrease post-traumatic stress disorder, depression and anxiety to increase the quality of life for patients following traffic injuries.


Biodegradation of di-n-butyl phthalate by bacterial consortium LV-1 enriched from river sludge.
A stable bacterial consortium (LV-1) capable of degrading di-n-butyl phthalate (DBP) was enriched from river sludge. Community analysis revealed that the main families of LV-1 are Brucellaceae (62.78%) and Sinobacteraceae (14.83%), and the main genera of LV-1 are Brucella spp. (62.78%) and Sinobacter spp. (14.83%). The optimal pH and temperature for LV-1 to degrade DBP were pH 6.0 and 30°C, respectively. Inoculum size influenced the degradation ratio when the incubation time was < 24 h. The initial concentration of DBP also influenced the degradation rates of DBP by LV-1, and the degradation rates ranged from 69.0-775.0 mg/l/d in the first 24 h. Degradation of DBP was best fitted by first-order kinetics when the initial concentration was < 300 mg/l. In addition, Cd2+, Cr6+, and Zn2+ inhibited DBP degradation by LV-1 at all considered concentrations, but low concentrations of Pb2+, Cu2+, and Mn2+ enhanced DBP degradation. The main intermediates (mono-ethyl phthalate [MEP], mono-butyl phthalate [MBP], and phthalic acid [PA]) were identified in the DBP degradation process, thus a new biochemical pathway of DBP degradation is proposed. Furthermore, LV-1 also degraded other phthalates with shorter ester chains (DMP, DEP, and PA).


Intermediate Charcot-Marie-Tooth disease: an electrophysiological reappraisal and systematic review.
Charcot-Marie-Tooth disease (CMT) is the most frequent form of inherited neuropathy with great variety of phenotypes, inheritance patterns, and causative genes. According to median motor nerve conduction velocity (MNCV), CMT is divided into demyelinating (CMT1) with MNCV below 38 m/s, axonal (CMT2) with MNCV above 38 m/s, and intermediate CMT with MNCV between 25 and 45 m/s. In each category, transmission may be autosomal dominant, autosomal recessive, or X-linked. The nosology of intermediate CMT is controversial because of concerns about electrophysiological delimitation. A systematic computer-based literature search was conducted on PubMed, using the following MeSH: (1) intermediate Charcot-Marie-Tooth; (2) X-linked intermediate Charcot-Marie-Tooth; and (3) X-linked Charcot-Marie-Tooth and electrophysiology. We retrieved 225 articles reporting X-linked CMT or intermediate CMT with electrophysiological information. After eligibility, 156 papers were used for this review. In assessing median MNCV, compound muscle action potential (CMAP) amplitudes were taken into account. In cases with attenuated CMAP and wherever possible, proximal median MNCV was used for accurate definition of conduction slowing in the intermediate range. In the vast majority of males with X-linked CMT associated with GJB1 mutation (CMTX1), median MNCV was intermediate. CMT associated with DRP2 mutation is another well-documented X-linked intermediate disorder. Autosomal dominant intermediate CMT (DI-CMT) encompasses 11 different types; six of them with assigned phenotype MIM number and the remaining five being unnumbered. Based on available electrophysiological information, we wonder if DI-CMTA should be reclassified within CMT2. Autosomal recessive intermediate CMT (RI-CMT) covers four numbered MIM phenotypes though, in accordance with reported electrophysiology, two of them (RI-CMTB and RI-CMTD) should probably be reclassified within AR-CMT2. We conclude that intermediate CMT is a complex inherited syndrome, whose characterization requires a specific electrophysiological protocol comprising evaluation of upper limb proximal nerve trunks when distal CMAP amplitudes are reduced, and that an updated version of MIM phenotype numbering is needed.


Vancomycin derivatives that inhibit peptidoglycan biosynthesis without binding D-Ala-D-Ala.
Vancomycin is an important drug for the treatment of Gram-positive bacterial infections. Resistance to vancomycin has begun to appear, posing a serious public health threat. Vancomycin analogs containing modified carbohydrates are very active against resistant microorganisms. Results presented here show that these carbohydrate derivatives operate by a different mechanism than vancomycin; moreover, peptide binding is not required for activity. It is proposed that carbohydrate-modified vancomycin compounds are effective against resistant bacteria because they interact directly with bacterial proteins involved in the transglycosylation step of cell wall biosynthesis. These results suggest new strategies for designing glycopeptide antibiotics that overcome bacterial resistance.


Health promotion: the emerging frontier in nursing.
Health promotion is gaining recognition as a health care strategy. The major premise of this paper is that the nature of health promotion is emerging. Rising into view, health promotion offers many challenges and opportunities to all health care professionals. Many of the influences that health promotion will have on nursing have not yet been fully explored. What is health promotion? What concepts are inherent in health promotion? Are there models of health promotion that are specific to nursing? What will be nursing's role in health promotion? What changes are required by the nursing profession to enable nurses to emerge within this new frontier? These are central questions for nurses as health promotion professionals to ponder, and attempts are made in this paper to answer these questions.


Defective 2D silicon phosphide monolayers for the nitrogen reduction reaction: a DFT study.
Electroreduction of N2 is a highly promising route for NH3 production. The lack of efficient catalysts that can activate and then reduce N2 into NH3 limits this as a pragmatic application. In this work, a 2D layered group IV-V material, silicon phosphide (SiP), is evaluated as a suitable substrate for the electrochemical nitrogen reduction reaction (ENRR). To capture N2, one phosphorus (P) defect was introduced on the plane of SiP. DFT calculations found that the defective SiP monolayer (D1-SiP, which is defined by the P-defect on SiP) exhibits enormous prospects towards the ENRR because of enhanced electron conductivity, good activation on N2, lower limiting potential (UL = -0.87 V) through the enzymatic pathway, smooth charge transfer between the catalyst and the reaction species, and robust thermal stability. Importantly, D1-SiP demonstrates the suppressed activities on producing of H2 and N2H4 side-products. This research demonstrates the potential of 2D metal-free Si-based catalysts for nitrogen fixation and further enriches the study of group IV-V materials for the ENRR.


Effects of Valsalva Maneuver on Ocular Biometric Parameters: Optical Low-Coherence Reflectometry Biometer Study.
To evaluate the effects of the Valsalva Maneuver (VM) on ocular biometry and intraocular pressure (IOP). This prospective observational study included 55 eyes of 55 healthy volunteers. Axial length (AL), central corneal thickness (CCT), anterior chamber depth (ACD), lens thickness (LT), corneal curvature (K-steep, K-flat and K-mean), corneal diameter (WtW), and pupil diameter (PD) were measured with a LenStar LS 900® biometer at rest and during VM. The IOP was also measured at rest and during VM measurements were compared. The VM did not have any significant influence on AL, WtW, K-flat, and K-mean (p > 0.05), but it decreased K-steep significantly (p < 0.001). Moreover, CCT and ACD decreased significantly during VM (p < 0.001), but VM increased IOP, PD (p < 0.001), and LT significantly (p = 0.002). The VM might reversibly change in IOP and ocular biometry, so it should be considered during anterior segment examinations.


Structural organization of the 16S ribosomal RNA from E. coli. Topography and secondary structure.
Extensive studies in our laboratory using different ribonucleases resulted in valuable data on the topography of the E.coli 16S ribosomal RNA within the native 30S subunit, within partially unfolded 30S subunits, in the free state, and in association with individual ribosomal proteins. Such studies have precise details on the accessibility of certain residues and delineated highly accessible RNA regions. Furthermore, they provided evidence that the 16S rRNA is organized in its subunit into four distinct domains. A secondary structure model of the E.coli 16S rRNA has been derived from these topographical data. Additional information from comparative sequence analyses of the small ribosomal subunit RNAs from other species sequenced so far has been used.


Identification of unknown quaternary ammonium compounds in corneal epithelium and aqueous humor.
Bovine corneal epithelium and bovine aqueous humor are investigated for their content of quaternary ammonium compounds. In total, four compounds are found. Three of these are identified. For the fourth compound, a proposal for its structure is made on the basis of tandem mass spectrometry fragmentation spectra. The compounds investigated have m/z values of 146, 160, and 174. The compounds with m/z 146 are confirmed as acetylcholine (in corneal epithelium) and (3-carboxypropyl)-trimethylammonium (in both corneal epithelium and aqueous humor). The compound with m/z 174 is identified as butyrylcholine (in corneal epithelium). The compound with m/z 160 is probably acetyl-g-homocholine (in both corneal epithelium and aqueous humor). For both butyrylcholine and acetyl-g-homocholine, it is the first time the presence of these compounds in corneal epithelium or aqueous humor (or both) is described. Both acetylcholine and butyrylcholine are unstable compounds, which are probably susceptible to enzymatic degradation by acetylcholine-esterase and butytrylcholine- esterase, respectively.


Expression and Function of Hypoxia Inducible Factor-1α and Vascular Endothelial Growth Factor in Pulp Tissue of Teeth under Orthodontic Movement.
Orthodontic force may lead to cell damage, circulatory disturbances, and vascular changes of the dental pulp, which make a hypoxic environment in pulp. In order to maintain the homeostasis of dental pulp, hypoxia will inevitably induce the defensive reaction. However, this is a complex process and is regulated by numerous factors. In this study, we established an experimental animal model of orthodontic tooth movement to investigate the effects of mechanical force on the expression of VEGF and HIF-1α in dental pulp. Histological analysis of dental pulp and expressions of HIF-1α and VEGF proteins in dental pulp were examined. The results showed that inflammation and vascular changes happened in dental pulp tissue in different periods. Additionally, there were significant changes in the expression of HIF-1α and VEGF proteins under orthodontic force. After application of mechanical load, expression of HIF-1α and VEGF was markedly positive in 1, 3, 7 d, and 2 w groups, and then it weakened in 4 w group. These findings suggested that the expression of HIF-1α and VEGF was enhanced by mechanical force. HIF-1α and VEGF may play an important role in retaining the homeostasis of dental pulp during orthodontic tooth movement.


Influence of Parental Exposure to Risk Factors in the Occurrence of Oral Clefts.
Non-syndromic cleft lip and palate are the most frequent craniofacial abnormalities in humans. The genetic, environmental and behavioral factors involved in this malformation must be clarified in different parts of the globe in the view of implementing preventive measures. To analyze the influence of parental exposure to risk factors on the occurrence of oral clefts. A case-control study was conducted with 150 mothers of oral cleft children paired by the children's gender to 300 mothers of children without congenital anomalies from Mato Grosso, Brazil, for the study of the variables: gender and race/color of the children; parental educational level; age; number of pregnancies; prenatal care; obesity; stress; diabetes; hypertension; use of medications, alcohol and illicit drugs; smoking and exposure to ionizing radiation during the first trimester of pregnancy. The results were analyzed in relation to the chances possibility of each variable for the occurrence of oral cleft through the bivariate and multivariate analysis by applying the model of logistic regression. Passive smoking, obesity, exposure to ionizing radiation and use of antibiotics were associated with the presence of clefts. The use of folic acid and analgesics were identified as preventive factors. The father's low educational level was found as a risk factor, while the black race/color was a preventive factor; nevertheless these variables were not associated in the multivariate analysis. The results reinforce the need to follow up the pregnant women, especially in the first trimester of pregnancy, in order to control the identified risk factors. Considering the factors associated with occurrence of oral clefts and those associated with its prevention, it is possible to apply specific health promotion measures during pregnancy, which can result in the reduction of oral clefts' occurrence.


Structure and antigenic behaviour of kappa I-immunoglobulin light-chain amyloid proteins.
N-terminal amino acid sequence analysis of three amyloid fibril subunit proteins from three different patients revealed a primary structure homologous to KI immunoglobulin light chains. In double immunodiffusion an antiserum against one of these amyloid proteins reacted with all three amyloids showing one line of identity, while an antiserum against one of the other amyloid proteins reacted with only one of the nonhomologous amyloids, appearing as partial identity, These two antisera did not react with any amyloid of lambda I, lambda IV or lambda VI type. The results confirm previous experiments, in which it has been shown that some antisera against amyloid proteins of immunoglobulin origin give a reaction of identity with other amyloids in the same immunoglobulin subgroup, probably owing to subgroup specific antigenic determinants in the variable segments. It was found that the distribution of amyloid varied widely among the three patients. There does not seem to be any correlation between the type of manifestation of amyloidosis and the type of immunoglobulin light chain forming the amyloid fibril.


Isolated hepatic perfusion with tumor necrosis factor and melphalan for unresectable cancers confined to the liver.
To evaluate the efficacy and systemic and regional toxicities of hyperthermic isolated hepatic perfusion (IHP) using tumor necrosis factor (TNF) and melphalan for the treatment of unresectable primary or metastatic cancers confined to the liver. Thirty-four patients (18 men and 16 women; mean age, 49 years) underwent a 60-minute hyperthermic (39.5 degrees to 40.0 degrees C) IHP performed by laparotomy that used TNF 1.0 mg and melphalan 1.5 mg/kg. Perfusion inflow was through the gastroduodenal artery and outflow was from a cannula positioned in an isolated segment of retrohepatic inferior vena cava (IVC). Infrahepatic IVC and portal venous blood flow were shunted to the axillary vein using an external venoveno bypass circuit. Complete vascular isolation of the liver was confirmed by an I-131-labelled human serum albumin monitoring technique. There was no operative mortality. Seventy-five percent of patients had reversible grade III or IV (National Cancer Institute Common Toxicity Criteria) hepatic toxicity with one treatment-related mortality (3%) because of hepatic venoocclusive disease. In 33 assessable patients, the overall response rate was 75% (complete response, one patient [3%]; partial response, 26 patients [72%]). With a median potential follow-up of 15 months, the mean duration of response was 9 months (range, 2 to 30 months). IHP with TNF and melphalan results in significant regression of bulky hepatic cancers confined to the liver in the majority of patients. Based on these initial results, further refinement of this treatment technique is warranted; perhaps by the combination of IHP with other regional treatment strategies to provide long-term control of unresectable cancers confined to liver.


Role of activation-induced deaminase protein kinase A phosphorylation sites in Ig gene conversion and somatic hypermutation.
Activation-induced deaminase (AID) is thought to initiate somatic hypermutation (SHM), gene conversion (GCV), and class switch recombination (CSR) by the transcription-coupled deamination of cytosine residues in Ig genes. Phosphorylation of AID by protein kinase A (PKA) and subsequent interaction of AID with replication protein A (RPA) have been proposed to play important roles in allowing AID to deaminate DNA during transcription. Serine 38 (S38) of mouse AID is phosphorylated in vivo and lies in a consensus target site for PKA, and mutation of this residue interferes with CSR and SHM. In this study, we demonstrate that S38 in mouse and chicken AID is phosphorylated in chicken DT40 cells and is required for efficient GCV and SHM in these cells. Paradoxically, zebra fish AID, which lacks a serine at the position corresponding to S38, has previously been shown to be active for CSR and we demonstrate that it is active for GCV/SHM. Aspartate 44 (D44) of zebra fish AID has been proposed to compensate for the absence of the S38 phosphorylation site but we demonstrate that mutation of D44 has no effect on GCV/SHM. Some features of zebra fish AID other than D44 might compensate for the absence of S38. Alternatively, the zebra fish protein might function in a manner that is independent of PKA and RPA in DT40 cells, raising the possibility that, under some circumstances, AID mediates efficient Ig gene diversification without the assistance of RPA.


'Snake-like' appearance of nuclear chromatin in conjunctival epithelial cells from patients with keratoconjunctivitis sicca.
Millipore filter imprints of conjunctiva in patients with conjunctival symptoms revealed a 'snake-like' appearance of the chromatin in nuclei of epithelial cells. A total of 40 patients were investigated. Abnormal cells were found in 8/16 patients with long-term keratoconjunctivitis sicca and in 6/24 patients suspected for Sjøgrens syndrome. The abnormal cells were typically arranged in clusters among morphologically normal epithelial cells. The finding was significantly correlated to the severity of the disease in patients with keratoconjunctivitis sicca. In all patients the abnormal cells could only be detected in millipore imprints of the upper bulbar conjunctiva and not in scrapings from the same area. The significance of the findings are discussed.


Oral immunization with recombinant listeria monocytogenes controls virus load after vaginal challenge with feline immunodeficiency virus.
Recombinant Listeria monocytogenes has many attractive characteristics as a vaccine vector against human immunodeficiency virus (HIV). Wild-type and attenuated Listeria strains expressing HIV Gag have been shown to induce long-lived mucosal and systemic T-cell responses in mice. Using the feline immunodeficiency virus (FIV) model of HIV we evaluated recombinant L. monocytogenes in a challenge system. Five cats were immunized with recombinant L. monocytogenes that expresses the FIV Gag and delivers an FIV Env-expressing DNA vaccine (LMgag/pND14-Lc-env). Control cats were either sham immunized or immunized with wild-type L. monocytogenes (LM-wt). At 1 year after vaginal challenge, provirus could not be detected in any of the nine tissues evaluated from cats immunized with the recombinant bacteria but was detected in at least one tissue in 8 of 10 control animals. Virus was isolated from bone marrow of four of five LMgag/pND14-Lc-env-immunized cats by use of a stringent coculture system but required CD8(+) T-cell depletion, indicating CD8(+) T-cell suppression of virus replication. Control animals had an inverted CD4:CD8 ratio in mesenteric lymph node and were depleted of both CD4(+) and CD8(+) intestinal epithelial T cells, while LMgag/pND14-Lc-env-immunized animals showed no such abnormalities. Vaginal FIV-specific immunoglobulin A was present at high titer in three LMgag/pND14-Lc-env-immunized cats before challenge and in all five at 1 year postchallenge. This study demonstrates that recombinant L. monocytogenes conferred some control of viral load after vaginal challenge with FIV.


FpvA receptor involvement in pyoverdine biosynthesis in Pseudomonas aeruginosa.
Alignment of the Pseudomonas aeruginosa ferric pyoverdine receptor, FpvA, with similar ferric-siderophore receptors revealed that the mature protein carries an extension of ca. 70 amino acids at its N terminus, an extension shared by the ferric pseudobactin receptors of P. putida. Deletion of fpvA from the chromosome of P. aeruginosa reduced pyoverdine production in this organism, as a result of a decline in expression of genes (e.g., pvdD) associated with the biosynthesis of the pyoverdine peptide moiety. Wild-type fpvA restored pvd expression in the mutant, thereby complementing its pyoverdine deficiency, although a deletion derivative of fpvA encoding a receptor lacking the N terminus of the mature protein did not. The truncated receptor was, however, functional in pyoverdine-mediated iron uptake, as evidenced by its ability to promote pyoverdine-dependent growth in an iron-restricted medium. These data are consistent with the idea that the N-terminal extension plays a role in FpvA-mediated pyoverdine biosynthesis in P. aeruginosa.


The accuracy of the swallowing kinematic analysis at various movement velocities of the hyoid and epiglottis.
To evaluate the accuracy of the swallowing kinematic analysis. To evaluate the accuracy at various velocities of movement, we developed an instrumental model of linear and rotational movement, representing the physiologic movement of the hyoid and epiglottis, respectively. A still image of 8 objects was also used for measuring the length of the objects as a basic screening, and 18 movie files of the instrumental model, taken from videofluoroscopy with different velocities. The images and movie files were digitized and analyzed by an experienced examiner, who was blinded to the study. The Pearson correlation coefficients between the measured and instrumental reference values were over 0.99 (p<0.001) for all of the analyses. Bland-Altman plots showed narrow ranges of the 95% confidence interval of agreement between the measured and reference values as follows: 0.14 to 0.94 mm for distances in a still image, -0.14 to 1.09 mm/s for linear velocities, and -1.02 to 3.81 degree/s for angular velocities. Our findings demonstrate that the distance and velocity measurements obtained by swallowing kinematic analysis are highly valid in a wide range of movement velocity.


Brahma regulates the Hippo pathway activity through forming complex with Yki-Sd and regulating the transcription of Crumbs.
The Hippo signaling pathway restricts organ size by inactivating the Yorkie (Yki)/Yes-associated protein (YAP) family proteins. The oncogenic Yki/YAP transcriptional coactivator family promotes tissue growth by activating target gene transcription, but the regulation of Yki/YAP activation remains elusive. In mammalian cells, we identified Brg1, a major subunit of chromatin-remodeling SWI/SNF family proteins, which interacts with YAP. This finding led us to investigate the in vivo functional interaction of Yki and Brahma (Brm), the Drosophila homolog of Brg1. We found that Brm functions at the downstream of Hippo pathway and interacts with Yki and Scalloped (Sd) to promotes Yki-dependent transcription and tissue growth. Furthermore, we demonstrated that Brm is required for the Crumbs (Crb) dysregulation-induced Yki activation. Interestingly, we also found that crb is a downstream target of Yki-Brm complex. Brm physically binds to the promoter of crb and regulates its transcription through Yki. Together, we showed that Brm functions as a critical regulator of Hippo signaling during tissue growth and plays an important role in the feedback loop between Crb and Yki.