Lysosome-Targeted Biosensor for the Super-Resolution Imaging of Lysosome-Mitochondrion Interaction.
Background: The interaction between lysosomes and mitochondria includes not only mitophagy but also mitochondrion-lysosome contact (MLC) that enables the two organelles to exchange materials and information. In our study, we synthesised a biosensor with fluorescence characteristics that can image lysosomes for structured illumination microscopy and, in turn, examined morphological changes in mitochondria and the phenomenon of MLC under pathological conditions. Methods: After designing and synthesising the biosensor, dubbed CNN, we performed an assay with a Cell Counting Kit-8 to detect CNN's toxicity in relation to H9C2 cardiomyocytes. We next analysed the co-localisation of CNN and the commercial lysosomal probe LTG in cells, qualitatively analysed the imaging characteristics of CNN in different cells (i.e. H9C2, HeLa and HepG2 cells) via structured illumination microscopy and observed how CNN entered cells at different temperatures and levels of endocytosis. Last, we treated the H9C2 cells with mannitol or glucose to observe the morphological changes of mitochondria and their positions relative to lysosomes. Results: After we endocytosed CNN, a lysosome-targeted biosensor with a wide, stable pH response range, into cells in an energy-dependent manner. SIM also revealed that conditions in high glucose induced stress in lysosomes and changed the morphology of mitochondria from elongated strips to round spheres. Conclusion: CNN is a new tool for tracking lysosomes in living cells, both physiologically and pathologically, and showcases new options for the design of similar biosensors.


Design of symmetric-sweep spectral-spatial RF pulses for spectral editing.
Spectral-spatial RF (SSRF) pulses allow simultaneous selection in both frequency and spatial domains. These pulses are particularly important for clinical and research MR spectroscopy (MRS) applications for suppression of large water and lipid resonances. Also, the high bandwidth of the subpulses (5-10 kHz) greatly reduces the spatial-shift errors associated with different chemical shifts. However, the use of high-bandwidth subpulses along with enough spectral bandwidth to measure a typical range of metabolite frequencies (e.g., 300 Hz at 3 T) can require RF amplitudes beyond the limits of the RF amplifier of a typical scanner. In this article, a new method is described for designing nonlinear-phase 180 degrees SSRF pulses that can be used for spectral editing. The novel feature of the pulses is that the spectral profile develops as a symmetric sweep, from the outside edges of the spectral window towards the middle, so that coupled components are tipped simultaneously and over a short interval. Pulses were designed for lactate editing at 1.5 T and 3 T. The spectral and spatial spin-echo profiles of the new pulses were measured experimentally. Spectra acquired in phantom experiments showed a well-resolved, edited lactate doublet, with 91% to 93% editing efficiency.


Single port entry - are there any advantages?
Minimal-invasive, single-port laparoscopic surgery is a recent innovation that may improve surgical outcomes. In this chapter, we review published research on single-port surgery in gynaecology, and the different surgical instruments available. Challenges, advantages, indications and potential future rules of this new approach are also discussed. Sixty-five studies were available for review: 17 case reports, 32 case studies, 13 retrospective comparative studies, and three randomised-controlled trials (RCTs). The recent availability of advanced instruments has made single-port surgery safer and more feasible for most benign gynaecologic surgeries. Single-port surgery has many potential benefits, but comparative trials have found no differences between single-port surgery and conventional laparoscopy in postoperative complications, postoperative pain, hospital stay, and cosmetic results. Single-port surgery seems to provide another option in the area of minimal invasive surgery, and further development of this technique, along with robotics and natural orifice transgastric endoscopic surgery, will improve dissemination of this approach.


Oct-based interpretation of the vitreomacular interface and indications for pharmacologic vitreolysis.
To review the role of optical coherence tomography (OCT) in diagnosis and management of vitreomacular disease and the impact of OCT on potential uses of ocriplasmin, a new pharmacologic vitreolysis agent recently approved by the U.S. Food and Drug Administration for the treatment of symptomatic vitreomacular adhesion. Analysis of current literature regarding OCT in diagnosis and management of vitreomacular interface disease. Posterior vitreous detachment is typically a nonpathologic age-related event. Anomalous posterior vitreous detachment emerges when the vitreous cortex fails to cleanly detach from the macula, optic nerve, or other adherent sites. Focal vitreomacular adhesion is a nonpathologic anatomical designation associated with perifoveal posterior vitreous detachment but normal retinal morphology on OCT. Vitreomacular traction is a pathologic consequence of persistent vitreous attachment with structural disturbance of the macular retina visible on OCT. Full-thickness macular holes are foveal defects continuous through all retinal layers to the retinal pigment epithelium. Vitreomacular traction and macular hole with focal vitreomacular adhesion are indications for pharmacologic vitreolysis. Noninvasive high-resolution OCT imaging has transformed the understanding of vitreomacular interface disease. Careful evaluation of the vitreomacular interface with OCT has increased in importance with the introduction of ocriplasmin for vitreomacular adhesion associated with symptomatic anatomical retinal changes.


Biomimetic Dextran-Based Hydrogel Layers for Cell Micropatterning over Large Areas Using the FluidFM BOT Technology.
Micropatterning of living single cells and cell clusters over millimeter-centimeter scale areas is of high demand in the development of cell-based biosensors. Micropatterning methodologies require both a suitable biomimetic support and a printing technology. In this work, we present the micropatterning of living mammalian cells on carboxymethyl dextran (CMD) hydrogel layers using the FluidFM BOT technology. In contrast to the ultrathin (few nanometers thick in the dry state) CMD films generally used in label-free biosensor applications, we developed CMD layers with thicknesses of several tens of nanometers in order to provide support for the controlled adhesion of living cells. The fabrication method and detailed characterization of the CMD layers are also described. The antifouling ability of the CMD surfaces is demonstrated by in situ optical waveguide lightmode spectroscopy measurements using serum modeling proteins with different electrostatic properties and molecular weights. Cell micropatterning on the CMD surface was obtained by printing cell adhesion mediating cRGDfK peptide molecules (cyclo(Arg-Gly-Asp-d-Phe-Lys)) directly from aqueous solution using microchanneled cantilevers with subsequent incubation of the printed surfaces in the living cell culture. Uniquely, we present cell patterns with different geometries (spot, line, and grid arrays) covering both micrometer and millimeter-centimeter scale areas. The adhered patterns were analyzed by phase contrast microscopy and the adhesion process on the patterns was real-time monitored by digital holographic microscopy, enabling to quantify the survival and migration of cells on the printed cRGDfK arrays.


Motor and Cognitive Developmental Profiles in Children With Down Syndrome.
To investigate motor and cognitive developmental profiles and to evaluate the correlation between two developmental areas and assess the influencing factors of the developmental process in children with Down syndrome (DS). Seventy-eight children with DS participated in this study. The medical history was taken and motoric milestone achievements recorded. The Korean Wechsler Preschool and Primary Scale of Intelligence (K-WPPSI) and Bayley Scales of Infant Development-II (BSID-II) were administered. Subjects were divided into severe motor delay group (severe group) and typical motor delay group (typical group). Children with DS follow the same sequence of motor development and generally displayed double times of acquisition of developmental milestones compared with healthy children. Furthermore, having surgery for associated complications showed negative influence to the motor development. Almost of all children with DS showed moderate degree of intellectual disability and motor and cognitive development do not seem to correlate one another. Surgery of associated complications can be negatively related to motor development. However, early motor development did not have any significant effects on the achievement of later cognitive functioning.


Succession and replacement of bacterial populations in the caecum of egg laying hens over their whole life.
In this study we characterised the development of caecal microbiota in egg laying hens over their commercial production lifespan, from the day of hatching until 60 weeks of age. Using pyrosequencing of V3/V4 variable regions of 16S rRNA genes for microbiota characterisation, we were able to define 4 different stages of caecal microbiota development. The first stage lasted for the first week of life and was characterised by a high prevalence of Enterobacteriaceae (phylum Proteobacteria). The second stage lasted from week 2 to week 4 and was characterised by nearly an absolute dominance of Lachnospiraceae and Ruminococcaceae (both phylum Firmicutes). The third stage lasted from month 2 to month 6 and was characterised by the succession of Firmicutes at the expense of Bacteroidetes. The fourth stage was typical for adult hens in full egg production aged 7 months or more and was characterised by a constant ratio of Bacteroidetes and Firmicutes formed by equal numbers of the representatives of both phyla.


Screening for loneliness in representative population samples: Validation of a single-item measure.
Loneliness is a highly relevant public mental health issue. This work presents the validation of a single-item measure of loneliness and its subjective experience: "I am frequently alone/have few contacts". It can be used in large-scale population surveys where an economical assessment is of key importance. Data was drawn from two representative German population surveys conducted in early and late 2020 (combined N = 4,984; 52.9% women; age: M = 48.39 years (SD = 17.88)). We determined the prevalence of loneliness in men and women across different age groups. In order to test concurrent validity, bivariate correlation analyses and Chi-square tests were performed. Convergent and discriminant validity were tested by investigating intercorrelations of the single-item measure of loneliness with another loneliness measure, other mental health outcomes, and associations with sociodemographic characteristics. Based on the single-item measure, 23.4% of participants reported some degree of loneliness, 3.4% among them severe loneliness. Comparisons with the LS-S showed similar prevalence rates of loneliness. A moderately positive relationship between the two loneliness measures was found by bivariate correlation analysis (ρ = .57, p < .001), but results indicated only weak convergent validity. Construct validity was supported by associations with depressive symptoms, anxiety symptoms, satisfaction with life, household size, and partnership. Loneliness is frequently reported in the general population. The single-item measure of loneliness is suitable as a brief screening measure in population-based assessments.


What has preoperative radio(chemo)therapy brought to localized rectal cancer patients in terms of perioperative and long-term outcomes over the past decades? A systematic review and meta-analysis based on 41,121 patients.
We asked what preoperative radiotherapy/chemoradiotherapy (PRT/PCRT) has brought to patients in terms of perioperative and long-term outcomes over the past decades. A systematic review and meta-analysis was conducted using PubMed, Embase and Web of Science databases. All original comparative studies published in English that were related to PRT/PCRT and surgical resection and which analyzed survival, postoperative and quality of life outcomes were included. Data synthesis and statistical analysis were carried out using Stata software. Data from 106 comparative studies based on 80 different trials enrolling 41,121 patients were included in our study. Based on our overall analyses, PRT/PCRT significantly improved patients' local recurrence-free survival (LRFS), but neither overall survival (OS) nor metastasis-free survival (MFS) showed improvement. In addition, PRT significantly increased the postoperative morbidity and mortality but PCRT did not have a significant effect. Furthermore, PRT/PCRT significantly increased the risk of postoperative wound complications but not anastomotic leakage and bowel obstruction. Our comprehensive subgroup analyses further supported the aforementioned results. Meanwhile, long-term anorectal symptoms (impaired squeeze pressures, use of pads, incontinence and urgency) and erectile dysfunction were also significantly increased in patients after PRT/PCRT. The benefits of PRT/PCRT as applied over the last several decades have not been sufficient to improve OS. Metastases of primary tumor and postoperative adverse effects were the two primary obstacles for an improved OS. In fact, the greatest advantage of PRT/PCRT is still local tumor control and a significantly improved LRFS.


Structural and ultrastructural changes in the developmental process of premature infants' and children's corneas.
Seven corneas ranging in age from 20 weeks of gestation to 6 months after birth were studied and changes observed in five layers. The epithelium thickens through the addition of new cellular layers and the enlargement of cells. The way junctional complexes develop with age is described. The way the collagen fibrils in Bowman's layer is arranged changes over time. The layer arrangement and orientation of the stroma's collagen fibrils are described at different stages of development. The keratocytes progressively flatten out and gather together. The thickness of Descemet's membrane increases from 0.6 m at 20 weeks of gestation to 4 m at 6 months. During the prenatal period, the banded layer appears with cross-linking bridges. The nonbanded layer appears after birth. Several changes were found in the endothelium: flattening out of the initially cube-shaped cells and changes in the intercellular junctions that can be seen from 20 weeks of fetal life. At 20 and 25 weeks of gestation, two layers of cells were observed in the corneal midperiphery. The adult structure of the cornea was reached at about 6 months after birth.


Aryl hydrocarbon receptor affects circadian-regulated lipolysis through an E-Box-dependent mechanism.
An internal circadian clock regulates timing of systemic energy homeostasis. The central clock in the hypothalamic suprachiasmatic nucleus (SCN) directs local clocks in peripheral tissues such as liver, muscle, and adipose tissue to synchronize metabolism with food intake and rest/activity cycles. Aryl hydrocarbon receptor (AhR) interacts with the molecular circadian clockworks. Activation of AhR dampens rhythmic expression of core clock genes, which may lead to metabolic dysfunction. Given the importance of appropriately-timed adipose tissue function to regulation of energy homeostasis, this study focused on mechanisms by which AhR may influence clock-controlled adipose tissue activity. We hypothesized that AhR activation in adipose tissue would impair lipolysis by dampening adipose rhythms, leading to a decreased lipolysis rate during fasting, and subsequently, altered serum glucose concentrations. Levels of clock gene and lipolysis gene transcripts in mouse mesenchymal stem cells (BMSCs) differentiated into mature adipocytes were suppressed by the AhR agonist β-napthoflavone (BNF), in an AhR dependent manner. BNF altered rhythms of core clock gene and lipolysis gene transcripts in C57bl6/J mice. BNF reduced serum free fatty acids, glycerol and liver glycogen. Chromatin immunoprecipitation indicated that BNF increased binding of AhR to E-Box elements in clock gene and lipolysis gene promoters. These data establish a link between AhR activation and impaired lipolysis, specifically by altering adipose tissue rhythmicity. In response to the decreased available energy from impaired lipolysis, the body increases glycogenolysis, thereby degrading more glycogen to provide necessary energy.


Upgrading of the STP Uithoorn: treatment of nutrient rich wastewater from horticulture.
The STP Uithoorn will be upgraded to accommodate the treatment of wastewater from a growing population and to meet more stringent nutrient discharge limits in 2006. In 2003 a system choice and preliminary design was made for the upgrading. A special feature is the nutrient rich wastewater flow from the rapidly developing horticulture in the area. Since the future loads from horticulture are highly uncertain, flexibility of the STP after upgrading is an important issue. A three stage system was selected: improved physical-chemical primary treatment, secondary treatment by activated sludge, and tertiary treatment by denitrifying filters. In this way an important part of the existing infrastructure can be reused, and flexibility is assured by constructing the tertiary treatment in modules and by providing a wide range of operational control possibilities. In this paper the process of system choice and selection of type of tertiary treatment are described, as well as the optimisation of the existing treatment. In order to determine the feasibility of allowing a high loading rate on the existing secondary clarifiers, a two-dimensional hydraulic model of the clarification process was used.


Quantum Monte Carlo calculations of the dissociation energy of the water dimer.
We report diffusion quantum Monte Carlo (DMC) calculations of the equilibrium dissociation energy D(e) of the water dimer. The dissociation energy measured experimentally, D(0), can be estimated from D(e) by adding a correction for vibrational effects. Using the measured dissociation energy and the modern value of the vibrational energy Mas et al., [J. Chem. Phys. 113, 6687 (2000)] leads to D(e)=5.00+/-0.7 kcal mol(-1), although the result Curtiss et al., [J. Chem. Phys. 71, 2703 (1979)] D(e)=5.44+/-0.7 kcal mol(-1), which uses an earlier estimate of the vibrational energy, has been widely quoted. High-level coupled cluster calculations Klopper et al., [Phys. Chem. Chem. Phys. 2, 2227 (2000)] have yielded D(e)=5.02+/-0.05 kcal mol(-1). In an attempt to shed new light on this old problem, we have performed all-electron DMC calculations on the water monomer and dimer using Slater-Jastrow wave functions with both Hartree-Fock approximation (HF) and B3LYP density functional theory single-particle orbitals. We obtain equilibrium dissociation energies for the dimer of 5.02+/-0.18 kcal mol(-1) (HF orbitals) and 5.21+/-0.18 kcal mol(-1) (B3LYP orbitals), in good agreement with the coupled cluster results.


Vascular Mapping for Abdominal-Based Breast Reconstruction: A Comprehensive Review of Current and Upcoming Imaging Modalities.
Preoperative vascular imaging is a very common element of surgical planning for abdominal-based breast reconstruction (ABBR). Surgeons must tailor which flap is best suited for each respective patient based on the patient's health and vascular anatomy. The goal of this review is to give surgeons practical tools for choosing which imaging technology best suits their patient's needs for successful breast reconstruction. A review of literature was undertaken on Google scholar to assess preoperative imaging modalities used for ABBR. Search terms included breast reconstruction, deep inferior epigastric perforator (DIEP) flap, and abdominal imaging. Articles were included based on relevance and significance to ABBR. Advantages and disadvantages of each imaging modality were then classified according to clinically relevant utility. Overall, imaging technologies that produce 3-dimensional images were found to have greater resolution for identifying perforators and the pedicle network than 2- dimensional images. This paper addresses the strengths and weaknesses of the currently used imaging modalities described and also discusses new technologies that may be helpful in the future for planning of ABBR.


Bone marrow microenvironment of patients with systemic lupus erythematosus.
In an attempt to clarify the hematologic abnormality in systemic lupus erythematosus (SLE), we studied the bone marrow microenvironment of 8 women with this disease. The number of fibroblast colonies was similar to that of healthy control individuals. Culture supernatants of monocytes obtained from the patients with SLE showed a diminished production of the hemopoietic growth factor required to stimulate bone marrow fibroblasts. As the activity of the humoral factor released from SLE monocytes was diminished, such a reduction may play a role in the pathogenesis of the hematologic abnormality seen in SLE.


Enhanced emission in organic nanocrystals via asymmetrical design of spirocyclic aromatic hydrocarbons.
Two spirocyclic aromatic hydrocarbon derivatives were prepared to clarify the molecular geometry effects on the regulation of the crystalline morphologies and photophysical behaviors of organic nanocrystals. Due to the different structural symmetry of a spiro-center, distinguishing nanocrystal morphologies with unique crystallization-enhanced/quenched emission was achieved.


DNA replication-dependent induction of gene proximity by androgen.
The male hormone androgen, working through the androgen receptor (AR), plays a major role in physiological process and disease development. Previous studies of AR mainly focus on its transcriptional activity. Here, we found that androgen-induced TMPRSS2 and ERG gene proximity is mediated by AR control of DNA replication rather than gene transcription. We demonstrate that, in both AR transactivation-positive and -negative prostate cells, androgen regulates DNA replication and androgen-induced gene proximity relies on both DNA replication-licensing and actual DNA replication activity. Androgen stimulation advances DNA replication timing of certain genomic regions, which may potentially increase gene proximity through sharing the same replication factory at a similar time. Therefore, we have revealed novel mechanisms of AR biological function, which will stimulate new research directions.


An in vivo membrane density perturbation strategy for identification of liver sinusoidal surface proteome accessible from the vasculature.
Liver sinusoidal endothelial cells (LSEC), the predominant nonparenchyma cells in liver, play critical roles in many important physiological and pathological processes by virtue of their unique location at the blood-tissue interface. To uncover the protein composition of LSEC plasma membrane (PM) comprehensively and give implications for the tissue microenvironment heterogeneity, we have developed an in vivo modified membrane density perturbation method for purification of the PM fraction. The proteins were separated and identified by SDS-PAGE combined with LC-MS/MS (GeLC-MS/MS). A total of 837 nonredundant proteins were identified, including a number of proteins previously reported to be localized to the PM of LSEC, as well as others not described. A diversity of membrane proteins involved in signaling, traffic, transporting and adhesion functions were identified. Our results demonstrated that the in vivo membrane density perturbation was an effective strategy to purify LSEC PM.


Human papillomavirus associated with vaginal intraepithelial neoplasia in women exposed to diethylstilbestrol in utero.
Five out of 959 young women, exposed to diethylstilbestrol (DES) in utero, developed vaginal intraepithelial neoplasia while they were under follow-up in the Diethylstilbestrol-Adenosis Project at Baylor College of Medicine, Houston, Texas. We suggest that the development of the vaginal intraepithelial neoplasia at a younger age than usual may be caused by a higher susceptibility of the DES-exposed patient to factors associated with the development of intraepithelial neoplasia. A common finding in all five women was the detection of the deoxyribonucleic acid (DNA) of human papillomavirus types 6 or 16 in their lesions, using high-stringency in situ hybridization. The role of human papillomavirus and herpes simplex virus in the etiology of intraepithelial neoplasia is discussed. Close follow-up is recommended for DES-exposed patients, especially those who have risk factors known to be associated with genital neoplasia.


Mechanisms by Which Pleiotropic Amphiphilic n-3 PUFA Reduce Colon Cancer Risk.
Colorectal cancer is one of the major causes of cancer-related mortality in both men and women worldwide. Genetic susceptibility and diet are primary determinants of cancer risk and tumor behavior. Experimental, epidemiological, and clinical data substantiate the beneficial role of n-3 polyunsaturated fatty acids (PUFA) in preventing chronic inflammation and colon cancer. From a mechanistic perspective, n-3 PUFA are pleiotropic and multifaceted with respect to their molecular mechanisms of action. For example, this class of dietary lipid uniquely alters membrane structure/ cytoskeletal function, impacting membrane receptor function and downstream signaling cascades, including gene expression profiles and cell phenotype. In addition, n-3 PUFA can synergize with other potential anti-tumor agents, such as fermentable fiber and curcumin. With the rising prevalence of diet-induced obesity, there is also an urgent need to elucidate the link between chronic inflammation in adipose tissue and colon cancer risk in obesity. In this review, we will summarize recent developments linking n-3 PUFA intake, membrane alterations, epigenetic modulation, and effects on obesity-associated colon cancer risk.


Applying the transactional stress and coping model to sickle cell disorder and insulin-dependent diabetes mellitus: identifying psychosocial variables related to adjustment and intervention.
This review paper examines the literature on psychosocial factors associated with adjustment to sickle cell disease and insulin-dependent diabetes mellitus in children through the framework of the transactional stress and coping (TSC) model. The transactional stress and coping model views adaptation to a childhood chronic illness as mediated by several psychosocial factors. This review examines the utility of the model in explaining adjustment in two different childhood diseases, identifies needed research and intervention targets, as well as highlights potential changes to the model. The major conclusions of this review suggest that, in addition to child-specific factors, family functioning is an area that interventions should address in sickle cell disease and insulin-dependent diabetes mellitus.


THE POPULATION GENETICS OF ADAPTATION: THE DISTRIBUTION OF FACTORS FIXED DURING ADAPTIVE EVOLUTION.
We know very little about the genetic basis of adaptation. Indeed, we can make no theoretical predictions, however heuristic, about the distribution of phenotypic effects among factors fixed during adaptation nor about the expected "size" of the largest factor fixed. Study of this problem requires taking into account that populations gradually approach a phenotypic optimum during adaptation via the stepwise substitution of favorable mutations. Using Fisher's geometric model of adaptation, I analyze this approach to the optimum, and derive an approximate solution to the size distribution of factors fixed during adaptation. I further generalize these results to allow the input of any distribution of mutational effects. The distribution of factors fixed during adaptation assumes a pleasingly simple, exponential form. This result is remarkably insensitive to changes in the fitness function and in the distribution of mutational effects. An exponential trend among factors fixed appears to be a general property of adaptation toward a fixed optimum.


Natural variation of ZmLNG1 alters organ shapes in maize.
The development of a series of elite maize hybrids has greatly increased crop yield in the past decades. Parental lines of these hybrids usually come from different heterotic groups and contain many genetic differences. Identifications of important quantitative trait genes in the elite hybrids can extend our understanding of heterosis and also help to guide genetic improvement. Here, we mapped a major quantitative trait locus using a linkage population from an elite maize hybrid Zhengdan958 and identified ZmLNG1 as the causative gene controlling multiple morphologic traits in maize. A 6-kb deletion in one parental line of the hybrid leads to the fusion of ZmLNG1 with its nearby gene. The fusion event prevents the C-terminal of ZmLNG1 from interacting with ZmTON1, which resulted in the change of plant architecture. Further experiments demonstrated that ZmLNG1 could act as a mediator to connect ZmTON1 and ZmOFPs, which belong to another type of plant morphological regulatory proteins, thereby affecting the phosphorylation level of ZmOFPs. These results demonstrate the importance of ZmLNG1 in forming the TON1-TRM-PP2A complex and provide a model for the regulation of plant organ morphology by TON1-recruiting motifs (TRMs) and Ovate family proteins (OFPs).


Management of thai cobra bites with a single bolus of antivenin.
Between January 1981 and December 1991, 68 adults were treated at Chulalongkorn University for cobra bites. Respiratory failure developed in the bite victims even when an adequate genus-specific antivenin was administered at the onset of neuromuscular paralysis. The duration of time spent on a respirator was, however, significantly less in patients treated with an intravenous 100-ml antivenin bolus given at the onset of neurotoxic signs. We found that a bolus of 100 ml of purified equine antivenin produced by the Queen Saovabha Memorial Institute, Bangkok, from Naja kauthia venom was adequate and that increasing the dose did not lead to added benefits. Time spent on a respirator was approximately 10 h in antivenin-treated groups and 40 h if no antivenin or an inadequate dose was used. Competent respiratory and nursing care alone without antivenin will lead to eventual recovery and prevent death. Local bite site tissue necrosis and duration of hospitalization were not related to the dose of antivenin given. Hospitalization was prolonged in subjects who developed sepsis or who had severe tissue necrosis or other surgical or medical complications.


Evidence Favoring a Positive Feedback Loop for Physiologic Auto Upregulation of hnRNP-E1 during Prolonged Folate Deficiency in Human Placental Cells.
Background: Previously, we determined that heterogeneous nuclear ribonucleoprotein E1 (hnRNP-E1) functions as an intracellular physiologic sensor of folate deficiency. In this model, l-homocysteine, which accumulates intracellularly in proportion to the extent of folate deficiency, covalently binds to and thereby activates homocysteinylated hnRNP-E1 to interact with folate receptor-α mRNA; this high-affinity interaction triggers the translational upregulation of cell surface folate receptors, which enables cells to optimize folate uptake from the external milieu. However, integral to this model is the need for ongoing generation of hnRNP-E1 to replenish homocysteinylated hnRNP-E1 that is degraded.Objective: We searched for an interrelated physiologic mechanism that could also maintain the steady-state concentration of hnRNP-E1 during prolonged folate deficiency.Methods: A novel RNA-protein interaction was functionally characterized by using molecular and biochemical approaches in vitro and in vivo.Results: l-homocysteine triggered a dose-dependent high-affinity interaction between hnRNP-E1 and a 25-nucleotide cis element within the 5'-untranslated region of hnRNP-E1 mRNA; this led to a proportionate increase in these RNA-protein complexes, and translation of hnRNP-E1 both in vitro and within placental cells. Targeted perturbation of this RNA-protein interaction either by specific 25-nucleotide antisense oligonucleotides or mutation within this cis element or by small interfering RNA to hnRNP-E1 mRNA significantly reduced cellular biosynthesis of hnRNP-E1. Conversely, transfection of hnRNP-E1 mutant proteins that mimicked homocysteinylated hnRNP-E1 stimulated both cellular hnRNP-E1 and folate receptor biosynthesis. In addition, ferrous sulfate heptahydrate [iron(II)], which also binds hnRNP-E1, significantly perturbed this l-homocysteine-triggered RNA-protein interaction in a dose-dependent manner. Finally, folate deficiency induced dual upregulation of hnRNP-E1 and folate receptors in cultured human cells and tumor xenografts, and more selectively in various fetal tissues of folate-deficient dams.Conclusions: This novel positive feedback loop amplifies hnRNP-E1 during prolonged folate deficiency and thereby maximizes upregulation of folate receptors in order to restore folate homeostasis toward normalcy in placental cells. It will also functionally impact several other mRNAs of the nutrition-sensitive, folate-responsive posttranscriptional RNA operon that is orchestrated by homocysteinylated hnRNP-E1.


Tubeless tracheotomy for survival airway surgery in the leporine model.
The ideal animal experimental tracheostomy technique is one that is 1) safe and easy to perform, 2) requires no tracheostomy tube, and 3) requires minimal cleaning or suctioning to maintain patency. The leporine model for airway injury has been well established and offers an inexpensive and practical animal model for experimental evaluation. However, previous research has demonstrated a high mortality rate with survival airway surgery in rabbits. This study demonstrates the feasibility of airway management in the leporine model using a simple maturing suture tracheostomy that avoids a tracheostomy tube. Tracheostomy was performed in six New Zealand white rabbits in the setting of survival surgery over a 2-week study period. A vertical tracheal incision was made from the second to the sixth tracheal ring. The anterior portion of the tracheal rings was removed and the skin surrounding the stoma was sutured down to the tracheal wall. The lateral tracheal wall was then suspended to the soft tissue in the lateral neck. All six rabbits survived the study period with minimal care and maintained stoma patency until sacrifice. Granulation tissue and edema were noted during the first week and largely resolved by the second week. An average of 5-mm-diameter stoma was measured 14 days after surgery without intraluminal stenosis or laryngeal edema. This method meets the defined criteria for the ideal experimental tracheostomy, demonstrating potential benefit in a laryngotracheal stenosis model and a rabbit model of evoked phonation.


Studies of respiratory morbidity in rubber workers. Part IV. Respiratory morbidity in talc workers.
Pulmonary function tests, chest x-rays, and respiratory questionnaires were administered to eighty talc workers and 189 non-exposed rubber workers from three rubber tire manufacturing plants. The talc workers, who were exposed to talc at levels below the current threshold limit value (TLV) of 20 mppcf for nonfibrous talc, had a statistically significantly greater prevalence of productive cough and of positive criteria for chronic obstructive lung disease (COLD) than did the control workers. The talc workers with more than 10 years of exposure had significantly decreased residual FEV 1.0. Multiple regression analysis of FEV 1.0 in the talc workers estimated that each year of exposure to talc dust reduced the FEV 1.0 by 26 ml. Talc workers had a clear increase in respiratory morbidity, despite the absence of chest roentgenographic changes. Based on this study, a safe exposure level for talc appears to be 25 mg/m3 as a time-weighted average.


Influence of family history on the willingness of outpatients to undergo genetic testing for salt-sensitive hypertension: a cross-sectional study.
It is unclear whether family medical history influences the willingness to undergo genetic testing. This study aimed to determine how family history affected the willingness to undergo genetic testing for salt-sensitive hypertension in patients with and without hypertension. Cross-sectional study using a self-administered questionnaire. Six primary care clinics and hospitals in Japan. Consecutive 1705 outpatients aged >20 years, 578 of whom had hypertension. The primary outcome variable was the willingness to undergo genetic testing to determine the risk of salt-sensitive hypertension, and the secondary variables were age, sex, education level, family history and concerns about hypertension. Factors associated with a willingness to undergo genetic testing were evaluated in patients with and without hypertension using a logistic regression model. In the hypertension and non-hypertension groups, 323 (55.9%) and 509 patients (45.2%), respectively, were willing to undergo genetic testing. This willingness was related with a high level of education (adjusted OR (ad-OR): 1.81, 95% CI 1.12 to 2.93), family history of stroke (1.55, 1.04 to 2.31) and concerns about hypertension (2.04, 1.27 to 3.28) in the hypertension group, whereas in the non-hypertension group, it was influenced by education level (ad-OR: 1.45, 95% CI 1.13 to 1.86), family history of hypertension (1.52, 1.17 to 1.98) and concerns about hypertension (2.03, 1.53 to 2.68). The influence of family history on the willingness to undergo genetic testing for risk of salt-sensitivity hypertension differed between participants with and without hypertension. In particular, participants without hypertension wished to know their likelihood of developing hypertension, whereas those with hypertension were interested to know the risk of stroke (a complication of hypertension). Family history could help better counsel patients about genetic testing on the basis of their medical history.


Determinants of access to physician services in Italy: a latent class seemingly unrelated probit approach.
We examine access to general practitioners and specialists who work in the public and private sectors in Italy using a seemingly unrelated system of probits. We use a latent class formulation that provides a rich and flexible functional form and can accommodate non-normality of response probabilities. The empirical analysis shows that patient behavior can be clustered in two latent classes. We find that income strongly influences the mix of services. Richer individuals are less likely to seek care from GP's and more likely to seek care from specialists, and especially private specialists. Health status and societal vulnerability are the most important indicators of class membership.


Diversity of human rotavirus VP6, VP7, and VP4 in Lagos State, Nigeria.
This study investigated the diversity of rotavirus strains recovered from young children in Lagos, Nigeria, during December 1996-January 1997. In total, 287 children, aged 1-60 month(s), presenting with diarrhoea to the Gbaja Health Centre of Massey Street Children Hospital and the Lagos University Teaching Hospital, were included in the study. Rotavirus-positive specimens were characterized by monoclonal antibody enzyme-linked immunosorbent assay (ELISA) for VP6 subgroup and VP7 serotype and by polymerase chain reaction (PCR) for VP4 genotype and VP7 strains (that were non-reactive to ELISA). Of 84 samples tested for VP6 subgroup epitope, subgroup II was predominant (51%) with only a few subgroup I strains (4%), while many could not be typed at all (45%). For the VP7 serotypes, G1 was the most prevalent strain (45%), followed by G3 strains (5%). Neither G2 nor G4 strains were found, although mixed G1/G2 has been reported for the first time in Nigeria. Of strains that were non-reactive to ELISA, 29 (34%) could not be typed by PCR for G type. A subset of 23 samples was selected on the basis of RNA electropherotype, VP7 serotype, and included nine strains of VP7 that were non-reactive to ELISA. VP4 genotype of this subset was determined by PCR, and the most prevalent genotype was P[6] (30%), followed by P[8] (26%). Only one P[4] strain was identified. This study has shown the diversity of rotavirus strains circulating in West Africa.


Effect of antisomatostatin gamma-globulin on gastrin release in rats.
In order to determine the mechanism of endogenous gastric somatostatin in the regulation of gastrin release, antisomatostatin rabbit gamma-globulin was administered in vivo and in vitro in the rat. First, in rats anesthetized by chloral hydrate, intravenous injection of 1 ml antisomatostatin rabbit gamma-globulin had no significant effect on plasma gastrin levels. Second, basal gastrin release from the isolated perfused rat stomach was not affected by the administration of antisomatostatin rabbit gamma-globulin (1:99 and 1:1 dilutions). Third, the addition of antisomatostatin rabbit gamma-globulin (1:99) to incubated rat antral mucosa, however, significantly increased the basal gastrin release. From these results, it is concluded that antral somatostatin exerts its inhibitory effect on basal gastrin release mainly not through the circulation but via paracrine pathways.


Inhibition of proto-oncogene c-fos transcription by inhibitors of protein kinase C and ion transport.
Both 4 beta-phorbol 12,13-didecanoate (PDD) and calcium ionophore A23187 induced c-fos mRNA accumulation with similar kinetics in human monocyte-like cells (U937). Their effects were additive. Cells pretreated with PDD were not induced to accumulate c-fos mRNA by PDD but remained responsive to the induction by A23187. Similarly cells pretreated with A23187 responded to PDD but not to A23187. Nuclear run-off transcription assay indicated that increase in the c-fos mRNA level after the second inducer treatment corresponded to transcriptional activation of the c-fos gene. The accumulation of c-fos mRNA and the transcriptional activation of c-fos induced by PDD or A23187 were inhibited by the protein kinase inhibitor H-7 and by quinidine and amiloride. The induction by A23187, but not that by PDD, was also inhibited by 4-aminopyridine, or tetraethylammonium. From these results, it is proposed that activation of protein kinase C and Na+ or K+ transport are required for c-fos induction caused by PDD and A23187.


A synthetic luteinizing hormone releasing hormone vaccine. I. Conjugation and specificity trials in BALB/c mice.
The immunobiology of luteinizing hormone releasing hormone (LHRH) was explored, to provide a conceptual and practical basis for the use of LHRH in immunocastration. Cysteine substituted analogues of LHRH were synthesized including Cys1-LHRH (C1-LHRH), Cys6-LHRH (C6-LHRH) and Cys10-LHRH (C10-LHRH). These were reacted to carrier molecules using the heterobifunctional cross-linking reagent m-maleimidobenzoylsulfosuccinimide ester (SMBS), producing peptide-carrier conjugates of known peptide content and conjugation orientation. This reaction regime was found to be rapid, efficient and allowed for easy control of peptide to carrier ratios. Conjugates were used in active immunization trials in BALB/c mice to characterize the murine immune response against LHRH. BALB/c mice were shown to have the capacity to recognise all three cysteine substituted LHRH analogues and to produce antibodies cross-reactive with native LHRH. The specificity of LHRH antisera generated was found to be dependent on the site of conjugation of the peptide to carrier molecule. C1-LHRH generated carboxy terminal directed antibodies, C10-LHRH generated amino terminal directed antibodies, while C6-LHRH could generate amino terminal directed or carboxy terminal directed antibodies, or both within a given animal. No intrinsically immunodominant epitopes were seen within the LHRH molecule.


Prognosis of early cervical cancer (FIGO Stages IA2, IB, and IIA) in northern Norway predicted by malignancy grading score and objective morphometric image analysis.
Recurrence of early-stage cervical cancer after primary surgery represents a considerable clinical problem, and, so far, few reliable markers for prediction of recurrence exist. Thus, the prognostic value of the malignancy grading score (MGS) classification system was evaluated in 82 patients with early-stage cervical cancer (International Federation of Gynecology and Obstetrics stages IA2, IB, and IIA) and long-time follow-up (5-16 years). Recurrence or not, the likelihood of lymph node metastases and reproducibility of the MGS semiquantitative system were tested. The prognostic power of the MGS to identify high-risk cases prone to recurrence in patients lacking lymph node metastases at primary surgery was a main purpose of the present study. The semiquantitative MGS classification system was performed independently by 2 pathologists unaware of prognosis and clinical data using light microscopy. Routine hematoxylin and eosin sections from surgical specimens were used, and investigation area was defined in the deep part of the tumor. Data-based image analysis was also used to investigate if objective morphometric parameters could add any prognostic power to MGS. The 5-year survival for the whole patient group was 92%. Malignancy grading score of greater than 17 risk points was statistically highly significant in predicting relapse and lymph node metastases (n = 82). High-risk cases lacking lymph node metastases (n = 70) were also statistically associated with high MGS. Depth of invasion and vascular invasion were statistically related to recurrence. Objective image analysis of nuclear parameters was of no additional statistical value for the prediction of outcome. The MGS classification system proved to be a useful tool in predicting recurrence and lymph node metastases and, most importantly, was a predictor of high-risk patients without metastases at primary surgery.


m6A Regulator-Mediated Methylation Modification Patterns and Characterisation of Tumour Microenvironment Infiltration in Non-Small Cell Lung Cancer.
The role of RNA N6-methyladenosine (m6A) modification in the progression of multiple tumours and the tumour microenvironment (TME) has been progressively demonstrated and promises a new direction for tumour therapy. However, there have been no reports on systematic analyses of RNA m6A modification in TME in non-small cell lung cancer (NSCLC). In this study, we used unsupervised cluster analysis to identify three m6A modification patterns of 28 m6A regulators and three m6A gene signature subgroups of commonly differentially expressed genes (co-DEGs) in the three m6A modification patterns. Quantifying these subtypes using the ssGSEA and ESTIMATE algorithms to characterise the tumour immune microenvironment (TIME) in NSCLC. Based on the principal component analysis (PCA), we used co-DEGs to construct m6A scores to analyse the characteristics of m6A modifications in individual patients and assessed the practical clinical utility of m6A scores using a nomogram for survival prediction. A total of 28 m6A regulators in 1210 NSCLC samples were mainly enriched in RNA modification and metabolic biological processes. The three following m6A modification patterns were identified based on the role of the 28 m6A regulators in TME: immune inflammation, immune evasion and immune desert. The m6A scores calculated based on co-DEGs in these modification patterns were significantly positively correlated with immune infiltration and significantly negatively correlated with tumour mutational burden (TMB). Survival was significantly better in the high-m6A-score group than in the low-m6A-score group, and the m6A score could be used as an independent favourable prognostic factor. In addition, assessment of both immune checkpoint inhibitors (ICIs) and immunophenoscore (IPS) revealed a better immunotherapeutic effect in the high-m6A-score group. The modification characteristics of 28 m6A regulators in the TIME of NSCLC were analysed from a comprehensive to an individual basis, which may facilitate the development of more effective clinical immunotherapeutic strategies.


Genotypic and phenotypic changes in exhaustively grown cell lines from mitochondrial cytopathy patients.
Understanding the pathobiology of mitochondrial (mt) DNA diseases involves both characterization of the effects of individual mutations on respiratory function and elucidation of the changes in mutation load and distribution (energy mosaicism) over serial cell generations. Whether a given mutation is stably maintained, or increases or decreases with cell growth, is one of the determinants as to whether a particular tissue will be affected by oxidative phosphorylation failure. In this study, we correlated mt genotype with biochemical phenotype in myoblasts from patients with pathogenic mtDNA mutations. The dominant process detected was a progressive elimination of mutant mtDNA genomes concomitant with an improvement in respiratory chain activity, suggesting that energetically normal cells have a growth advantage over those with a high mutation load. We propose that this elimination is by biased distribution of wild-type mtDNA to daughter cells, and that a similar mechanism could operate in vivo and contribute to both the clinical expression of mt disease and the maintenance of a predominantly wild-type mt genome pool across generations.


Attenuation of the electrophysiological function of the corpus callosum after fluid percussion injury in the rat.
This study describes a new method used to evaluate axonal physiological dysfunction following fluid percussion induced traumatic brain injury (TBI) that may facilitate the study of the mechanisms and novel therapeutic strategies of posttraumatic diffuse axonal injury (DAI). Stimulated compound action potentials (CAP) were recorded extracellularly in the corpus callosum of superfused brain slices at 3 h, and 1, 3, and 7 days following central fluid percussion injury and demonstrated a temporal pattern of functional deterioration. The maximal CAP amplitude (CAPA) covaried with the intensity of impact 1 day following sham, mild (1.0-1.2 atm), and moderate (1.8-2.0 atm) injury (p < 0.05; 1.11 +/- 0.10, 0.82 +/- 0.11, and 0.49 +/- 0.08 mV, respectively). The CAPA in sham animals were approximately 1.1 mV and did not vary with survival interval (3 h, and 1, 3, and 7 days); however, they were significantly decreased at each time point following moderate injury (p < 0.05; 0.51 +/- 0.11, 0.49 +/- 0.08, 0.46 +/- 0.10, and 0.75 +/- 0.13 mV, respectively). The CAPA at 7 days in the injured group were higher than at 3 h, and 1 and 3 days. H&E and amyloid precursor protein (APP) light microscopic analysis confirmed previously reported trauma-induced axonal injury in the corpus callosum seen after fluid percussion injury. Increased APP expression was confirmed using Western blotting showing significant accumulation at 1 day (IOD 913.0 +/- 252.7; n = 3; p = 0.05), 3 days (IOD 753.1 +/- 159.1; n = 3; p = 0.03), and at 7 days (IOD 1093.8 = 105.0; n = 3; p = 0.001) compared to shams (IOD 217.6 +/- 20.4; n = 3). Thus, we report the characterization of white matter axonal dysfunction in the corpus callosum following TBI. This novel method was easily applied, and the results were consistent and reproducible. The electrophysiological changes were sensitive to the early effects of impact intensity, as well as to delayed changes occurring several days following injury. They also indicated a greater degree of attenuation than predicted by APP expression changes alone.


Annular bullous lesions with atypical erythema multiforme in leprosy.
Erythema nodosum leprosum (ENL) is an immune complex-mediated reaction that may complicate the course of multibacillary leprosy. Bullous lesions in Type II reaction, though reported, are exceedingly rare. We report the case of a 32 year old female patient who presented initially at our OPD with erythema nodosum. Cutaneous examination revealed impaired sensation over dorsum of right foot and thickened right lateral popliteal nerve. Slit skin smear (SSS) from ear lobes revealed AFB with a bacteriological index of 2+. She was started on MDT, tablet ofloxacin 200 mg twice a day, and 30 mg oral prednisolone. Two months later, she presented with generalised pruritus, large target lesions over the back, and hemorrhagic bullae over lower extremities and annular pattern of bullae, over both arms. A SSS was repeated which was positive for AFB. Histopathology from bullous lesions was consistent with ENL. Direct Immunofluorescence (DIF) study was negative. Our patient improved rapidly after she was started on thalidomide 100 mg twice daily, with withdrawal of ofloxacin. Erythema Multiforme (EMF) and annular bullous lesions have been reported in patients on treatment with ofloxacin. This case is being presented due to the unusual and varied manifestation of Type II lepra reaction in a 34 year old female patient.


Estimating causal parameters without target populations.
In recent years, numerous research methodologists have argued forcefully that any estimated effect from an observational study or a randomized trial should apply to a 'target population'- to a finite group of people. Some methods to adjust for confounders heavily draw upon this idea. I cite a recently published paper in The American Journal of Epidemiology that linked methods to adjust for confounders to the concept of a 'target population'. I explain that the requirement to specify a finite population as the target of causal inference is rooted in two extreme models of causation: determinism and stochastic causation. I argue that the 'target population' epistemology is scientifically irrelevant and so are methods to handle confounders that are founded on this paradigm, namely, standardization, inverse-probability-of-treatment weighting and SMR-weighting. Finally, I propose a simple alternative framework under an indeterministic model of causation. According to my proposed model, a causal parameter is not tied to any finite population and its estimate is a (fallible) scientific conjecture about a homogeneous, individual-level effect.


Epoxy resin/carbon black composites below the percolation threshold.
A set of epoxy resin composites filled with 0.25-2.0 wt.% of commercially available ENSACO carbon black (CB) of high and low surface area (CBH and CBL respectively) has been produced. The results of broadband dielectric spectroscopy of manufactured CB/epoxy below the percolation threshold in broad temperature (200 K to 450 K) and frequency (20 Hz to 1 MHz) ranges are reported. The dielectric properties of composites below the percolation threshold are mostly determined by alpha relaxation in pure polymer matrix. The glass transition temperature for CB/epoxy decreases in comparison with neat epoxy resin due to the extra free volume at the polymer-filler interface. At room temperature, the dielectric permittivity is higher for epoxy loaded with CBH additives. In contrast, at high temperature, the electrical conductivity was found to be higher for composites with CBL embedded. The established influence of the CB surface area on the broadband dielectric characteristics can be exploited for the production of effective low-cost antistatic paints and coatings working at different temperatures.


Antibody responses to malarial antigens in the Wopkaimin population of the Star Mountains, Papua New Guinea.
Antibody responses to malarial antigens were determined in 614 serum samples collected from the Wopkaimin population of the Star Mountains of Papua New Guinea. In point prevalence surveys made in 1982-1983, 33.7% of the persons examined were infected with Plasmodium falciparum, P. vivax, or P. malariae. Of these, 72.9% were infected with P. falciparum. In a standard fluorescent antibody test, highest level responses were to P. falciparum, followed by P. malariae, P. vivax, and P. ovale. A strong correlation was found between results of the fluorescent antibody tests and those obtained in an enzyme-linked immunosorbent assay using P. falciparum antigens. The failure of immune responses to eliminate these species of Plasmodium in this highly isolated population is discussed.


Time autocorrelation function analysis of master equation and its application to atomic clusters.
We derive the energy fluctuation Delta(2)E, and the time autocorrelation kappa(tau) and its Fourier transformation--the fluctuation spectra S(omega)--of the master-equation transition matrix. The contribution from each eigenmode of the transition matrix to these fluctuation quantities reveals the relevant importance of the individual mode in the relaxation processes. The time scales associated with these relaxation processes are determined by the corresponding eigenvalues. Unlike traditional time evolution analysis, the autocorrelation function and fluctuation spectra analysis does not involve an arbitrary initial population. It is also more suitable for analyzing the underlying dynamic, kinetic behavior near the equilibrium and the behavior of the long-time-scale rare events. We utilize our technique to analyze the solid-liquid phase coexistence of the 13-atom Morse cluster and the fcc-to-icosahedral structure transition of the 38-atom Lennard-Jones cluster. For the processes studied, the fluctuation spectra from the master equation simplify the analysis of the transition matrix, and the important relaxation modes are easily extracted.


Prevalence and distribution of HSV-1, VZV, and HHV-6 in human cranial nerve nuclei III, IV, VI, VII, and XII.
The etiology of idiopathic cranial nerve palsies often remains unresolved. It has been hypothesised that viral reactivation of herpesviruses in the corresponding nuclei in the brainstem is the cause. We investigated the distribution of herpes simplex virus type 1 (HSV-1) and varicella zoster virus (VZV) in nuclei that are associated with peripheral sensory ganglia [oculomotor (nIII), facial (nVII) nuclei] and in nuclei that are not associated with peripheral sensory ganglia [trochlear (nIV), abducens (nVI), and hypoglossal (nXII) nuclei] of five human brainstems. Samples of the cranial nerve nuclei and adjacent control tissue were taken from histological sections after precise identification of every single nucleus and control tissue. DNA and RNA amplification methods were used to determine the prevalence and distribution of HSV-1 and VZV. The distribution of human herpes virus type 6 (HHV-6) was also determined and served as a control, since HHV-6 infection has never been associated with idiopathic cranial nerve palsies. HSV-1 was distributed at random in all cranial nerve nuclei and control tissue, whereas VZV DNA was not detected in any of the samples examined. Surprisingly, HHV-6 was present in almost all samples where HSV-1 was also present, however, the latency associated transcript (LAT) of HSV-1 was not found in any of the samples positive for HSV-1 DNA. The absence of LAT in the samples positive for HSV-1 and the distribution of HSV-1 and HHV-6 do not support the hypothesis that idiopathic cranial nerve palsies result from viral reactivation in the brainstem nuclei.


Enhancement of PPARα-Inhibited Leucine Metabolism-Stimulated β-Casein Synthesis and Fatty Acid Synthesis in Primary Bovine Mammary Epithelial Cells.
Leucine, a kind of branched-chain amino acid, plays a regulatory role in the milk production of mammalian mammary glands, but its regulatory functions and underlying molecular mechanisms remain unknown. This work showed that a leucine-enriched mixture (LEUem) supplementation increased the levels of milk protein and milk fat synthesis in primary bovine mammary epithelial cells (BMECs). RNA-seq of leucine-treated BMECs indicated alterations in lipid metabolism, translation, ribosomal structure and biogenesis, and inflammatory response signaling pathways. Meanwhile, the supplementation of leucine resulted in mTOR activation and increased the expression of BCKDHA, FASN, ACC, and SCD1. Interestingly, the expression of PPARα was independently correlated with the leucine-supplemented dose. PPARα activated by WY-14643 caused significant suppression of lipogenic genes expression. Furthermore, WY-14643 attenuated leucine-induced β-casein synthesis and enhanced the level of BCKDHA expression. Moreover, promoter analysis revealed a peroxisome-proliferator-response element (PPRE) site in the bovine BCKDHA promoter, and WY-14643 promoted the recruitment of PPARα onto the BCKDHA promoter. Together, the present data indicate that leucine promotes the synthesis of β-casein and fatty acid and that PPARα-involved leucine catabolism is the key target.


Study of Escherichia coli O114 associated with a hospital outbreak.
Escherichia coli O114 strains isolated from infants in the course of a hospital outbreak of enteritis were identical with type strain 26W in antigenic structure (O114:K90a, 90b), and in immunoelectrophoretic pattern (ORSKOV's group 1Bb) but differed from it in fermenting salicin late (18-22 days) and in phage pattern (group A4); the strains produced colicin and were lysogenic. Pathogenicity of the organism was proved by IgM type antibodies in a patient's serum rising in a titre of 1:128 and by its enterotoxin-producing capacity shown by the rabbit ileal loop assay.


Advancing Patient Care Through Specialty Pharmacy Services in an Academic Health System.
With the rapid growth of specialty pharmacies, including those within academic health systems, pharmacists have the opportunity to improve patient care through the management of specialty medications. Specialty pharmacists within academic health systems are uniquely positioned to overcome restrictions to medication access, financial constraints, and provider burdens that often lead to obstacles for patients to start and maintain necessary treatments. The Vanderbilt Specialty Pharmacy (VSP) model at Vanderbilt University Medical Center (VUMC) provides an example of a patient-centered, collaborative care prototype that places pharmacists directly into specialty clinics to assist with comprehensive management of patients on specialty medications. VSP integrates specialty pharmacy services within existing specialty clinics based on the needs of each individual clinic. Each clinic is staffed with at least 1 clinical pharmacist and 1 pharmacy technician. The pharmacist is integrally involved in medication selection, initiation, and monitoring. The specialty pharmacy team ensures appropriate medication access and cost, provides extensive medication education, ensures patients are adherent to treatment, and coordinates care between patients and providers using the electronic medical record. Integration of pharmacists within specialty clinics at VUMC benefits providers, the health system, and patient care. This model has demonstrated decreased provider and clinic burden, decreased time to medication approval and initiation, excellent patient and provider satisfaction, substantial patient cost savings, optimal medication adherence, and overall improved continuity of care for patients on specialty medications. Since its inception in 2011, VSP has integrated 24 clinical pharmacists and 17 pharmacy technicians into 20 specialty clinics, with continued quarterly growth. The VSP model advances the role of pharmacists in managing patients on specialty medications in collaboration with providers. The integrated collaborative approach as presented by VSP represents a best practices model for those establishing and advancing specialty pharmacy services within academic health systems. No outside funding supported this study. The authors have nothing to disclose. Study concept and design were principally contributed by Bagwell and Newman, along with the other authors. Lee took the lead in data collection, along with Carver, Bagwell, Kelley, and Newman. Data interpretation was performed by Carver, Kelley, Lee, and Bagwell, with assistance from Newman. The manuscript was written by Bagwell, Carver, Kelley, and Lee and revised primarily by Bagwell, along with the other authors.


A radioimmunoassay for parathymosin alpha using antibodies to synthetic N-terminal peptide 1-30.
Antibodies against the N-terminus of rat parathymosin alpha have been raised in rabbits by conjugating parathymosin alpha (1-30) to hemocyanin. A radioimmunoassay for parathymosin alpha was established by utilizing antibodies against the above polypeptide and parathymosin alpha(1-12)[Tyr] as tracer. The useful range was 5-450 pmol for parathymosin alpha. An epitope was located in the amino acid sequence 1-12. The antiserum failed to crossreact with the same molar concentrations of the partly homologous thymosin alpha 1 or prothymosin alpha. With this radioimmunoassay, parathymosin alpha was isolated from calf thymus after separation from prothymosin alpha by reversed phase HPLC. Endogenous proteases did not appear to generate N-terminal fragments of parathymosin alpha in rat liver extracts in a similar fashion to that observed for prothymosin alpha. Parathymosin alpha has a ubiquitous distribution in the human tissues examined, with levels ranging from 93 (brain) to 1043 (liver) ng of parathymosin alpha(1-30) equivalents/g (wet weight).


Biliary excretion of digoxin in man.
We have measured the biliary excretion of 3H-digoxin in normal subjects by intestinal perfusion methods which impose minimal interruption of the enterohepatic circulation. In contrast to data obtained in patients with liver disease and postoperative patients with biliary fistulas, our data show that about 30% of an intravenous dose of 3H-digoxin reaches the digestive tract in 24 hr. Most of the excreted radioactivity is chloroform-soluble and presumably reabsorbably and most of this fraction is probably biologically active drug. The results suggest that (1) consideration of hepatobiliary function may be relevant in observations regarding digoxin pharmacokinetics and that this route may be exploited in trials of therapy designed to shorten the duration of digoxin intoxication.


Regression models for expected length of stay.
In multi-state models, the expected length of stay (ELOS) in a state is not a straightforward object to relate to covariates, and the traditional approach has instead been to construct regression models for the transition intensities and calculate ELOS from these. The disadvantage of this approach is that the effect of covariates on the intensities is not easily translated into the effect on ELOS, and it typically relies on the Markov assumption. We propose to use pseudo-observations to construct regression models for ELOS, thereby allowing a direct interpretation of covariate effects while at the same time avoiding the Markov assumption. For this approach, all we need is a non-parametric consistent estimator for ELOS. For every subject (and for every state of interest), a pseudo-observation is constructed, and they are then used as outcome variables in the regression model. We furthermore show how to construct longitudinal (pseudo-) data when combining the concept of pseudo-observations with landmarking. In doing so, covariates are allowed to be time-varying, and we can investigate potential time-varying effects of the covariates. The models can be fitted using generalized estimating equations, and dependence between observations on the same subject is handled by applying the sandwich estimator. The method is illustrated using data from the US Health and Retirement Study where the impact of socio-economic factors on ELOS in health and disability is explored. Finally, we investigate the performance of our approach under different degrees of left-truncation, non-Markovianity, and right-censoring by means of simulation.


[The effect of physical training on lipid parameters in the blood].
Physical training leads to an improved metabolic capacity of musculature. At the same time through a decreased liberation of catecholamines a reduction of the increased lipolysis develops. The two factors together condition an improved glucose tolerance and a decrease of the reactive insulin secretion. Thus, among others, the synthesis of triglycerides is reduced and an essential factor of risk is favourably influenced for the development of arteriosclerosis.


Transcriptome comparison of distinct osteolineage subsets in the hematopoietic stem cell niche using a triple fluorescent transgenic mouse model.
The bone marrow niche is recognized as a central player in maintaining and regulating the behavior of hematopoietic stem and progenitor cells. Specific gain-of and loss-of function experiments perturbing a range of osteolineage cells or their secreted proteins had been shown to affect stem cell maintenance (Calvi et al, 2003 [1]; Stier et al., 2005 [2]; Zhang et al., 2003 [3]; Nilsson et al., 2005 [4]; Greenbaum et al., 2013 [5]) and engraftment (Adam et al., 2006, 2009 [6,7]). We used specific in vivo cell deletion approaches to dissect the niche cell-parenchymal cell dependency in a complex bone marrow microenvironment. Endogenous deletion of osteocalcin-expressing (Ocn(+)) cells led to a loss of T immune cells (Yu et al., 2015 [8]. Ocn(+) cells express the Notch ligand DLL4 to communicate with T-competent progenitors, and thereby ensuring T precursor production and expression of chemotactic molecules on their cell surface for subsequent thymic seeding. In contrast, depletion of osterix-expressing (Osx(+)) osteoprogenitors led to reduced B immune cells. These distinct hematopoietic phenotypes suggest specific pairing of mesenchymal niche cells and parenchymal hematopoietic cells in the bone marrow to create unique functional units to support hematopoiesis. Here, we present the global gene expression profiles of these osteolineage subtypes utilizing a triple fluorescent transgenic mouse model (OsxCre(+);Rosa-mCh(+);Ocn:Topaz(+)) that labels Osx(+) cells red, Ocn(+) cells green, and Osx(+) Ocn(+) cells yellow. This system allows isolation of distinct osteolineage subsets within the same animal by flow cytometry. Array data that have been described in our study [8] are also publically available from NCBI Gene Expression Omnibus (GEO) with the accession number GSE66042. Differences in gene expression may correlate with functional difference in supporting hematopoiesis.


Repurposing of tamoxifen ameliorates CLN3 and CLN7 disease phenotype.
Batten diseases (BDs) are a group of lysosomal storage disorders characterized by seizure, visual loss, and cognitive and motor deterioration. We discovered increased levels of globotriaosylceramide (Gb3) in cellular and murine models of CLN3 and CLN7 diseases and used fluorescent-conjugated bacterial toxins to label Gb3 to develop a cell-based high content imaging (HCI) screening assay for the repurposing of FDA-approved compounds able to reduce this accumulation within BD cells. We found that tamoxifen reduced the lysosomal accumulation of Gb3 in CLN3 and CLN7 cell models, including neuronal progenitor cells (NPCs) from CLN7 patient-derived induced pluripotent stem cells (iPSC). Here, tamoxifen exerts its action through a mechanism that involves activation of the transcription factor EB (TFEB), a master gene of lysosomal function and autophagy. In vivo administration of tamoxifen to the CLN7Δex2 mouse model reduced the accumulation of Gb3 and SCMAS, decreased neuroinflammation, and improved motor coordination. These data strongly suggest that tamoxifen may be a suitable drug to treat some types of Batten disease.


Electromagnetic field enhancement in the gap between two Au nanoparticles: the size of hot site probed by surface-enhanced Raman scattering.
We have estimated the apparent size of the 'hot' site for surface-enhanced Raman scattering (SERS) located within the gap between two spherical Au nanoparticles. Initially, 55-nm sized Au nanoparticles are laid on a thiol-group terminated silane film, and then 1,4-phenylenediisocyanide (1,4-PDI) molecules are self-assembled onto the Au nanoparticles. 1,4-PDI is bonded to Au by forming one Au-CN bond, with another isocyanide group being pendent with respect to the Au surface. Up to this point, no Raman scattering is detected at all for 1,4-PDI. Upon attaching new Au nanoparticles onto the pendent isocyanide groups of 1,4-PDI, a Raman signal is distinctly observed. In agreement with the finite-difference time-domain (FDTD) calculation, this is understood in terms of the electromagnetic hot sites formed at the gaps between two Au nanoparticles. The Raman signal does not increase further, however, even after the adsorption of additional 1,4-PDI onto the vacant surfaces of the second Au nanoparticles. To a rough estimate, about 400 molecules residing only within a 10-nm diameter area of the center of the gap, adsorbed in the first stage, are then conjectured to have contributed most of the measured Raman signal of 1,4-PDI. This apparently indicates that the size of 'hot' site is very limited.


Biofilm community succession: a neutral perspective.
Although biofilms represent one of the dominant forms of life in aqueous environments, our understanding of the assembly and development of their microbial communities remains relatively poor. In recent years, several studies have addressed this and have extended the concepts of succession theory in classical ecology into microbial systems. From these datasets, niche-based conceptual models have been developed explaining observed biodiversity patterns and their dynamics. These models have not, however, been formulated mathematically and so remain untested. Here, we further develop spatially resolved neutral community models and demonstrate that these can also explain these patterns and offer alternative explanations of microbial succession. The success of neutral models suggests that stochastic effects alone may have a much greater influence on microbial community succession than previously acknowledged. Furthermore, such models are much more readily parameterised and can be used as the foundation of more complex and realistic models of microbial community succession.


The Dorsal Medial Prefrontal Cortex Is Recruited by High Construal of Non-social Stimuli.
The dorsomedial prefrontal cortex (dmPFC) is part of the mentalizing network, a set of brain regions consistently engaged in inferring mental states. However, its precise function in this network remains unclear. It has recently been proposed that the dmPFC is involved in high-level abstract (i.e., categorical) identification or construction of both social and non-social stimuli, referred to as "high construal." This was based on the observation of greater activation in the dmPFC shared by a high construal social condition (trait inference based on visually presented behavior) and a high construal non-social condition (categorization of visually presented objects) vs. matched low construal conditions (visual description of the same pictures). However, dmPFC activation has been related to task contexts requiring responses based on self-guided generation of mental content or decisions as compared to responses more directly determined by the experimental context (e.g., free vs. rule-governed choice). The previously reported dmPFC activity may reflect differences in task constraint (i.e., the extent to which the task context guided the process) confounded with the construal manipulation. Therefore, in the present study, we manipulated construal level and constraint independently, while participants underwent functional magnetic resonance imaging (fMRI). As before, participants visually described (low level construal) or categorized (high level construal) pictures of objects. Orthogonal to this, the description or categorization task had to be performed on either one object (low constraint) or on two objects simultaneously (high constraint), limiting the number of possible responses. Statistical analysis revealed common greater activation in both high construal conditions (high and low constraint) than in their low construal counterparts, replicating the influence of construal level on dmPFC activation (greater involvement in high than low construal), but no influence of constraint. In line with previous proposals and earlier work, we suggest that the dmPFC is involved in high-construal abstraction across different domains.


Oral bioavailability of insulin contained in polysaccharide nanoparticles.
The pharmacological activity of insulin-loaded dextran sulfate/chitosan nanoparticles was evaluated following oral dosage in diabetic rats. Nanoparticles were mucoadhesive and negatively charged with a mean size of 500 nm, suitable for uptake within the gastrointestinal tract. Insulin association efficiency was over 70% and was released in a pH-dependent manner under simulated gastrointestinal conditions. Orally delivered nanoparticles lowered basal serum glucose levels in diabetic rats around 35% with 50 and 100 IU/kg doses sustaining hypoglycemia over 24 h. Pharmacological availability was 5.6 and 3.4% for the 50 and 100 IU/kg doses, respectively, a significant increase over 1.6%, determined for oral insulin alone in solution. Confocal microscopic examinations of FITC-labeled insulin nanoparticles showed adhesion to rat intestinal epithelium, and internalization of insulin within the intestinal mucosa. Encapsulation of insulin into dextran sulfate/chitosan nanoparticles was a key factor in the improvement of the bioavailability of its oral delivery over insulin solution.


Expression of receptor for advanced glycation end products (RAGE) and MMP-9 in human pancreatic cancer cells.
Amphoterin is considered as a regulator for the ability of invasion and migration in tumor cells and embryonic neurons through binding to receptor for advanced glycation end products (RAGE), a multiligand cell surface molecule of the immunoglobulin superfamily. As matrix metalloproteinase-9 (MMP-9, gelatinase B) has been reported to play a critical role in tumor progression and metastasis, we have examined the relation of RAGE and MMP in human pancreatic cancer. Three representative human pancreatic carcinoma cells were rendered for the study which show different metastatic potential, PANC-1 and MIA PaCa-2 as the cells with high ability, BxPC-3 as with low. The expression of RAGE was examined by RT-PCR. The expression of MMP-9 protein was examined by Western blotting. RAGE was strongly expressed in MIA PaCa-2 and PANC-1 that have high metastatic ability. On the contrary, RAGE was expressed little in BxPC-3 that has low ability. Similarly, expression of MMP-9 showed almost the same tendency. RAGE and MMP-9 are expressed concordant with the metastatic ability of the human pancreatic cancer cells. Control of these molecules could be a key to regulating the metastatic ability of pancreatic cancer and this may be exploited in targeted therapy of this cancer.


Improving the efficiency of breast radiotherapy treatment planning using a semi-automated approach.
To reduce treatment planning times while maintaining plan quality through the introduction of semi-automated planning techniques for breast radiotherapy. Automatic critical structure delineation was examined using the Smart Probabilistic Image Contouring Engine (SPICE) commercial autosegmentation software (Philips Radiation Oncology Systems, Fitchburg, WI) for a cohort of ten patients. Semiautomated planning was investigated by employing scripting in the treatment planning system to automate segment creation for breast step-and-shoot planning and create objectives for segment weight optimization; considerations were made for three different multileaf collimator (MLC) configurations. Forty patients were retrospectively planned using the script and a planning time comparison performed. The SPICE heart and lung outlines agreed closely with clinician-defined outlines (median Dice Similarity Coefficient > 0.9); median difference in mean heart dose was 0.0 cGy (range -10.8 to 5.4 cGy). Scripted treatment plans demonstrated equivalence with their clinical counterparts. No statistically significant differences were found for target parameters. Minimal ipsilateral lung dose increases were also observed. Statistically significant (P < 0.01) time reductions were achievable for MLCi and Agility MLC (Elekta Ltd, Crawley, UK) plans (median 4.9 and 5.9 min, respectively). The use of commercial autosegmentation software enables breast plan adjustment based on doses to organs at risk. Semi-automated techniques for breast radiotherapy planning offer modest reductions in planning times. However, in the context of a typical department's breast radiotherapy workload, minor savings per plan translate into greater efficiencies overall.


Ultrasonographic and computed tomography findings in renal suppurations: performance indicators and risks for diagnostic failure.
The main purpose of the study was to identify and quantify clinical variables which predisposed incorrect diagnosis based on ultrasonography (US) and computed tomography (CT) findings in the patients with suppurative renal infections. A retrospective review of radiological records of patients at 3 tertiary Serbian clinics of urology from 1999 to January 2006 was conducted. The series consisted of 49 women and 26 men, mean age = 56.7 years, ranging from 21 to 78. All patients underwent US examinations. Forty-one patients had undergone CT examinations, on 42 renal units (1 bilateral perirenal infection). The patients were categorized into 4 groups according to the type of suppurative infection: (a) unilocular renal or perirenal abscesses; (b) with pyonephrosis only; (c) with multiple renal or pyonephrosis infections along with extension on perirenal space, and (d) emphysematous infections. Incorrect diagnosis based on US record only depended on the suppurative entity. In the presence of pyonephrosis only, the risk of incorrect diagnosis was reduced for 4 times [odds ratio (OR) = 0.25], and in presence of emphysematous infections the risk increased 19.5 times (OR = 19.49). Perinephric abscesses which were not seen on US were those associated with pyonephrosis, abscesses smaller than 6 cm and gas-forming abscesses. The diagnosis of abscesses considerably varied in the cases of unilocular purulent collections compared to complex purulent collections (chi(2) = 8.177, p = 0.004). Overall, the risk for incorrect diagnosis is about 14 times higher using US only (OR = 14.5), while CT reduced the risk of it for about 37 times (OR = 0.027). CT was much more reliable than US to correctly identify the pathological entity of renal suppuration. Guiding treatment on US findings only seems hazardous as omission of some pathological processes is very likely.


Inpatient Communication Networks: Leveraging Secure Text-Messaging Platforms to Gain Insight into Inpatient Communication Systems.
This study attempts to characterize the inpatient communication network within a quaternary pediatric academic medical center by applying network analysis methods to secure text-messaging data. We used network graphing and statistical software to create network models of an inpatient communication system with secure text-messaging data from physicians, nurses, and other ancillary staff in an academic medical center. Descriptive statistics about the network, users within the network, and visualizations informed the team's understanding of the network and its components. Analysis of messages exchanged over approximately 23 days revealed a large, scale-free network with 4,442 nodes and 59,913 edges. Quantitative description of user behavior (messages sent and received) and network metrics (i.e., importance of nodes within a network) revealed several operational and clinical roles both sending and receiving > 1,000 messages over this time period. While some of these nodes represented expected "dispatcher" roles in our inpatient system, others occupied important frontline clinical roles responsible for bedside clinical care. Quantitative and network analysis of secure text-messaging logs revealed several key operational and clinical roles at risk for alert fatigue and information overload. This analysis also revealed a communication network highly reliant on these key roles, meaning disruption to these individuals or their workflows could lead to dysfunction of the communication network. While secure text-messaging applications play increasingly important roles in facilitating inpatient communication, little is understood about the impact these systems have on health care providers. Developing methods to understand and optimize communication between inpatient providers might help operational and clinical leaders to proactively prevent poorly understood pitfalls associated with these systems and build resilient and effective communication structures.


Metabolic and nutrition support in the chronic critical illness syndrome.
Technological innovations in the ICU have led to artificially prolonged life, with an associated cost. Chronic critical illness (CCI) occurs in patients with prolonged mechanical ventilation and allostatic overload, and is associated with a discrete and consistent metabolic syndrome. Metabolic interventions are extrapolated from clinical critical care research, scientific theory, and years of CCI patient care experience. Intensive metabolic support (IMS) is a multi-targeted approach consisting of tight glycemic control with intensive insulin therapy, early and adequate nutrition therapy, nutritional pharmacology, management of metabolic bone disease, and meticulous attention to other endocrine/metabolic derangements. Ideally, IMS should be under the supervision of a metabolic support consultative team. Further research specifically focused on the CCI population is needed to validate this current approach.


The roles of kisspeptin revisited: inside and outside the hypothalamus.
Kisspeptin, encoded by KISS1/Kiss1 gene, is now considered a master regulator of reproductive functions in mammals owing to its involvement in the direct activation of gonadotropin-releasing hormone (GnRH) neurons after binding to its cognate receptor, GPR54. Ever since the discovery of kisspeptin, intensive studies on hypothalamic expression of KISS1/Kiss1 and on physiological roles of hypothalamic kisspeptin neurons have provided clues as to how the brain controls sexual maturation at the onset of puberty and subsequent reproductive performance in mammals. Additionally, emerging evidence indicates the potential involvement of extra-hypothalamic kisspeptin in reproductive functions. Here, we summarize data regarding kisspeptin inside and outside the hypothalamus and revisit the physiological roles of central and peripheral kisspeptins in the reproductive functions of mammals.


Within country inequalities in caesarean section rates: observational study of 72 low and middle income countries.
To provide an update on economic related inequalities in caesarean section rates within countries. Secondary analysis of demographic and health surveys and multiple indicator cluster surveys. 72 low and middle income countries with a survey conducted between 2010 and 2014 for analysis of the latest situation of inequality, and 28 countries with a survey also conducted between 2000 and 2004 for analysis of the change in inequality over time. Women aged 15-49 years with a live birth during the two or three years preceding the survey. Data on caesarean section were disaggregated by asset based household wealth status and presented separately for five subgroups, ranging from the poorest to the richest fifth. Absolute and relative inequalities were measured using difference and ratio measures. The pace of change in the poorest and richest fifths was compared using a measure of excess change. National caesarean section rates ranged from 0.6% in South Sudan to 58.9% in the Dominican Republic. Within countries, caesarean section rates were lowest in the poorest fifth (median 3.7%) and highest in the richest fifth (median 18.4%). 18 out of 72 study countries reported a difference of 20 percentage points or higher between the richest and poorest fifth. The highest caesarean section rates and greatest levels of absolute inequality were observed in countries from the region of the Americas, whereas countries from the African region had low levels of caesarean use and comparatively lower levels of absolute inequality, although relative inequality was quite high in some countries. 26 out of 28 countries reported increases in caesarean section rates over time. Rates tended to increase faster in the richest fifth (median 0.9 percentage points per year) compared with the poorest fifth (median 0.2 percentage points per year), indicating an increase in inequality over time in most of these countries. Substantial within country economic inequalities in caesarean deliveries remain. These inequalities might be due to a combination of inadequate access to emergency obstetric care among the poorest subgroups and high levels of caesarean use without medical indication in the richest subgroups, especially in middle income countries. Country specific strategies should address these inequalities to improve maternal and newborn health.


ProMEX - a mass spectral reference database for plant proteomics.
The ProMEX database is one of the main collection of annotated tryptic peptides in plant proteomics. The main objective of the ProMEX database is to provide experimental MS/MS-based information for cell type-specific or sub-cellular proteomes in Arabidopsis thaliana, Medicago truncatula, Chlamydomonas reinhardtii, Lotus japonicus, Lotus corniculatus, Phaseolus vulgaris, Lycopersicon esculentum, Solanum tuberosum, Nicotiana tabacum, Glycine max, Zea mays, Bradyrhizobium japonicum, and Sinorhizobium meliloti. Direct links at the protein level to the most relevant databases are present in ProMEX. Furthermore, the spectral sequence information are linked to their respective pathways and can be viewed in pathway maps.


Quantitative genetics of costly neonatal sexual size dimorphism in squirrel monkeys (Saimiri boliviensis).
Offspring size is often an intimate link between the fitness of parents and offspring. Among mammals, neonate mass is also related to adult levels of dimorphism and intrasexual competitive mating. We describe the sex-specific genetic architecture of neonate mass in captive squirrel monkeys (Saimiri boliviensis), a small Neotropical primate. Best fitting quantitative genetic models show strong maternal genetic effects with little difference between sexes offering limited opportunity for neonatal dimorphism to respond to observed or hypothetical selection. Heritabilities that are approximately zero also imply it is unlikely that neonatal dimorphism can evolve as a correlated response to selection on adult size. However, male mass is also more dependent on maternal condition (age and parity) making dimorphism plastic. Finally, we hypothesize that large maternal genetic effects reflect income breeding and tightly synchronized seasonal reproduction in squirrel monkeys, both of which require strong maternal control of offspring growth and timing of birth.


Reliance on visible speech cues during multimodal language processing: individual and age differences.
The current study demonstrates that when a strong inhibition process is invoked during multimodal (auditory-visual) language understanding: older adults perform worse than younger adults, visible speech does not benefit language-processing performance, and individual differences in measures of working memory for language do not predict performance. In contrast, in a task that does not invoke inhibition: adult age differences in performance are not obtained, visible speech benefits language performance, and individual differences in working memory predict performance. The results offer support for a framework for investigating multimodal language processing that incorporates assumptions about general information processing, individual differences in working memory capacity, and adult cognitive aging.


Resveratrol-Enriched Rice Down-Regulates Melanin Synthesis in UVB-Induced Guinea Pigs Epidermal Skin Tissue.
Synthetic compounds that are used in the clinic to regulate skin hyperpigmentation, such as arbutin, hydroquinone, and kojic acid, are only moderately effective. But, their use is limited by side effects. As part of an effort to overcome the limitations, we developed resveratrol-enriched rice (RR) using genetic engineering technique. Each of resveratrol and rice has been reported to produce anti-melanogenic effects. Therefore, we hypothesized that RR would show more anti-melanogenic effects than those of resveratrol or rice alone. Anti-melanogenic effect of RR was done by using melan-a mouse melanocytes. The depigmenting efficacy was then observed following topical application of the RR to UVB-stimulated hyperpigmented dorsal skin of guinea pigs. Treatment with RR extract resulted a 21.4 ± 0.7% decrease in tyrosinase expression at melan-a cells. Colorimetric analysis showed a significantly lower depigmenting value by day 9 following treatment with RR in UVB-irradiated guinea pigs the dorsal skin (p<0.01), indicating that RR produced a depigmentation effect. By staining with Fontana-Masson stain, we found that the RR-treated group had more effect histopathologically in epidermal melanin production than resveratrol or rice alone-treated group. RR was associated with reduction in the levels of microphthalmia-associated transcription factor (MITF), and downregulation of tyrosinase and tyrosinase-related protein (TRP-2) expression, leading to inhibit epidermal melanin production by western blot analysis. This study suggests that the resveratrol-enriched rice may be a promising candidate in regulating skin pigmentation with UVB exposure.


Elder mistreatment: dental assessment and intervention.
Elder mistreatment has increasingly been recognized as a serious and complex health issue affecting large numbers of elders each year. Dependent elder populations are often at heightened risk for mistreatment by those individuals and institutions that care for them. Health professionals have been found to lack knowledge regarding assessment, diagnosis, intervention, and reporting criteria of elder mistreatment. This article reviews the types and prevalence of elder mistreatment, risk factors, signs and symptoms, preventive interventions, and ethical considerations.


Co-factor engineering in lactobacilli: effects of uncoupled ATPase activity on metabolic fluxes in Lactobacillus (L.) plantarum and L. sakei.
The hydrolytic F(1)-part of the F(1)F(0)-ATPase was over-expressed in Lactobacillus (L.) plantarum NC8 and L. sakei Lb790x during fermentation of glucose or ribose, in order to study how changes in the intracellular levels of ATP and ADP affect the metabolic fluxes. The uncoupled ATPase activity resulted in a decrease in intracellular energy level (ATP/ADP ratio), biomass yield and growth rate. Interestingly, the glycolytic and ribolytic flux increased in L. plantarum with uncoupled ATPase activity compared to the reference strain by up to 20% and 50%, respectively. The ATP demand was estimated to have approximately 80% control on both the glycolytic and ribolytic flux in L. plantarum under these conditions. In contrast, the glycolytic and ribolytic flux decreased in L. sakei with uncoupled ATPase activity.


Virtual Screening of 1,4-Naphthoquinone Derivatives for Inhibition of a Key Cancer Signaling Protein, AKT1 Kinase.
AKT, also known as protein kinase B (PKB), is an established therapeutic target in cancer and its inhibitors are increasingly designed. The anti-cancer potential of a compound class naphthoquinones has been constantly realized. The current work aimed to explore AKT1 inhibitors from 1,4-naphthoquionone derivatives. A library of 1,4-naphthoquionone derivatives was formed using similarity search and visual analysis. The library was used for virtual screening using molecular docking. For the screened compounds, the detailed binding pose analysis, binding energy and dissociation constant calculations were performed. The top 10 screened compounds were proposed as potential AKT1 inhibitors with anti-cancer activity. The compounds were checked for any reported activity, and our 2nd rank compound was reported to have anti-cancer activity. Our study proposes 10 compounds as potential AKT1 inhibitors and anticancer agents and also provides insights into their binding. This study also proposes AKT1 as a potential target of the reported anticancer compound, CID: 341807.


Long-Term Outcome of Immediate Versus Postponed Intervention in Patients With Infected Necrotizing Pancreatitis (POINTER): Multicenter Randomized Trial.
To compare the long-term outcomes of immediate drainage versus the postponed drainage approach in patients with infected necrotizing pancreatitis. In the randomized POINTER trial, patients assigned to the postponed-drainage approach using antibiotic treatment required fewer interventions, as compared to immediate drainage, and over a third were treated without any intervention. Clinical data of those patients alive after the initial 6-month follow-up were re-evaluated. Primary outcome was a composite of death and major complications. Out of 104 patients, 88 were re-evaluated with a median follow-up of 51 months. After the initial 6-month follow-up, the primary outcome occurred in 7 of 47 patients (15%) in the immediate-drainage group and 7 of 41 patients (17%) in the postponed-drainage group (RR 0.87, 95% CI 0.33-2.28; P =0.78). Additional drainage procedures were performed in 7 patients (15%) versus 3 patients (7%) (RR 2.03; 95% CI 0.56-7.37; P =0.34). The median number of additional interventions was 0 (IQR 0-0) in both groups ( P =0.028). In the total follow-up, the median number of interventions was higher in the immediate-drainage group than in the postponed-drainage group (4 vs. 1, P =0.001). Eventually, 14 of 15 patients (93%) in the postponed-drainage group who were successfully treated in the initial 6-month follow-up with antibiotics and without any intervention, remained without intervention. At the end of follow-up, pancreatic function and quality of life were similar. Also during long-term follow-up, a postponed drainage approach using antibiotics in patients with infected necrotizing pancreatitis results in fewer interventions as compared to immediate drainage, and should therefore be the preferred approach. ISRCTN33682933.