Effects of 1-methylcyclopropene and post-controlled atmosphere air storage treatments on fresh-cut Ambrosia apple slices.
The effect of 1-methylcyclopropene (1-MCP) treatment and two different post-controlled atmosphere air storage (PCAAS) durations on the quality and chemistry of fresh-cut Ambrosia apple slices was studied. PCAAS for 1 or 2 weeks prior to slicing had an overall positive effect on the resultant quality of fresh-cut apple slices. The most significant responses to PCAAS were the suppression of both phenolic and o-quinone accumulation in slices, and this was related to the significantly lower browning potential values obtained for slices from PCAAS-treated apples. Polyphenol oxidase (PPO), peroxidase (POX) and ascorbate peroxidase (APOX) activities were not affected by 1-MCP or PCAAS treatments. PPO and POX activities were almost completely inhibited by a 50 g L⁻¹ calcium ascorbate anti-browning dip of apple slices from all treatments. The most dramatic effect of the PCAAS treatments was to reduce the accumulation of soluble phenolics, which is likely the reason that o-quinone accumulation was also inhibited in treated fruits. The consequent reduction in browning potential may be the explanation as to why PCAAS treatment has been shown to reduce fresh apple slice browning in previous work.


Maternal deprivation induces depressive-like behaviour and alters neurotrophin levels in the rat brain.
The present study was aimed to evaluate the behavioral and molecular effects of maternal deprivation in adult rats. To this aim, male rats deprived and non-deprived were assessed in the forced swimming and open-field tests in adult phase. In addition adrenocorticotrophin hormone (ACTH) levels was assessed in serum and brain-derived-neurotrophic factor (BDNF), neurotrophin-3 (NT-3) and nerve growth factor (NGF) protein levels were assessed in prefrontal cortex, hippocampus and amygdala. We observed that maternal deprivation increased immobility time, and decreased climbing time, without affecting locomotor activity. ACTH circulating levels were increased in maternal deprived rats. Additionally, BDNF protein levels were reduced in the amygdala and NT-3 and NGF were reduced in both hippocampus and amygdala in maternal deprived rats, compared to control group. In conclusion, our results support the idea that behavioral, ACTH circulating levels and neurotrophins levels altered in maternal deprivation model could contribute to stress-related diseases, such as depression.


In vitro studies of plasmid-mediated penicillinase from Streptococcus faecalis suggest a staphylococcal origin.
A strain of Streptococcus faecalis with plasmid-mediated penicillinase production was studied further. Partially purified penicillinase from the S. faecalis strain hydrolyzed penicillin, ampicillin, and ureido-penicillins but not penicillinase-resistant semisynthetic penicillins, cephalosporins, or imipenem; hydrolysis was inhibited by clavulanic acid. Hydrolysis of a given antibiotic correlated with a marked increase in the minimal inhibitory concentration (MIC) of that drug when a high inoculum was used. As with most enterococci, the MICs of cephalosporins and penicillinase-resistant semisynthetic penicillins were too high for clinical usefulness, although these agents did not show an inoculum effect. Based upon hybridization under stringent conditions of plasmid DNA from the S. faecalis strain to cloned penicillinase genes from Staphylococcus aureus, it appears that these resistance determinants are highly homologous and suggests that this enzyme was introduced into streptococci from staphylococci.


When to err is inhuman: An examination of the influence of artificial intelligence-driven nursing care on patient safety.
Artificial intelligence, as a nonhuman entity, is increasingly used to inform, direct, or supplant nursing care and clinical decision-making. The boundaries between human- and nonhuman-driven nursing care are blurred with the advent of sensors, wearables, camera devices, and humanoid robots at such an accelerated pace that the critical evaluation of its influence on patient safety has not been fully assessed. Since the pivotal release of To Err is Human, patient safety is being challenged by the dynamic healthcare environment like never before, with nursing at a critical juncture to steer the course of artificial intelligence integration in clinical decision-making. This paper presents an overview of artificial intelligence and its application in healthcare and highlights the implications which affect nursing as a profession, including perspectives on nursing education and training recommendations. The legal and policy challenges which emerge when artificial intelligence influences the risk of clinical errors and safety issues are discussed.


Role of NF-κB pathway in kidney renal clear cell carcinoma and its potential therapeutic implications.
Kidney renal clear cell carcinoma (KIRC), a common malignant tumor of the urinary system, is the most aggressive renal tumor subtype. Since the discovery of nuclear factor kappa B (NF-κB) in 1986, many studies have demonstrated abnormal NF-κB signaling is associated with the development of various cancers, including kidney renal clear cell carcinoma. In this study, the relationship between NF-κB and kidney renal clear cell carcinoma was confirmed using bioinformatics analysis. First, we explored the differential expression of copy number variation (CNV), single nucleotide variant (SNV), and messenger RNA (mRNA) in NF-κB-related genes in different types of cancer, as well as the impact on cancer prognosis and sensitivity to common chemotherapy drugs. Then, we divided the mRNA expression levels of NF-κB-related genes in KIRC patients into three groups through GSVA cluster analysis and explored the correlation between the NF-κB pathway and clinical data of KIRC patients, classical cancer-related genes, common anticancer drug responsiveness, and immune cell infiltration. Finally, 11 tumor-related genes were screened using least absolute shrinkage and selection operator (LASSO) regression to construct a prognostic model. In addition, we used the UALCAN and HPA databases to verify the protein levels of three key NF-κB-related genes (CHUK, IKGGB, and IKBKG) in KIRC. In conclusion, our study established a prognostic survival model based on NF-κB-related genes, which can be used to predict the prognosis of patients with KIRC.


P072 Non-invasive Vagal Nerve Stimulation to Treat Crohn Disease and Ulcerative Colitis in Children and Young Adults: A Proof-of-Concept Clinical Trial.
Current inflammatory bowel disease (IBD) therapies are limited by incomplete efficacy, loss of response, side effects and high costs. Vagal nerve stimulation (VNS) is an investigational Bioelectronic Medicine therapy targeting the nervous system to treat IBD through an 'inflammatory reflex' that reduces systemic inflammation. Data in humans with IBD are promising but limited to two small studies using a cervically implanted VNS device in adults with Crohn disease (CD); no data exist to date using a non-invasive VNS modality in IBD. This study aimed to evaluate the efficacy and safety of transcutaneous auricular VNS (ta-VNS) in children and young adults with CD or ulcerative colitis (UC). IBD patients 10-21 years of age with mild/moderate CD or UC who did not achieve remission with conventional therapy, and a fecal calprotectin (FC) >200 ug/g within 4 weeks of study entry were enrolled. Subjects were randomized to receive either ta-VNS using a transcutaneous electrical nerve stimulator unit targeting the cymba conchae of the external left ear, or sham stimulation of the posterior lower leg, for 5 minutes once daily for a 2-week duration followed by a switch to the alternative stimulation for an additional 2 weeks. At week 4, all subjects were assigned to receive active ta-VNS for 5 minutes twice daily until week 16 so that all received 14 weeks of active ta-VNS by the end of the study. Primary study endpoint was defined as ∆FC ≥50% reduction from baseline to week 16. Secondary endpoints included improvement in weighted Pediatric Crohn Disease Activity Index (wPCDAI) or Pediatric Ulcerative Colitis Activity Index (PUCAI). Twenty-two subjects were enrolled (12 UC, 10 CD; median age 14.4 years [range 10-21] 54.5% male). Median baseline FC was 587 (range 18 - 3828, SD 1074). 5 subjects (4 UC, 1 CD) had FC levels 12.5 at baseline, 3 (50%) achieved clinical remission (wPCDAI ≤ 10) at week 16. In the full UC cohort (n = 12), baseline PUCAI ranged from 0-45. In the 6 with PUCAI > 10 at baseline, 2 (33%) achieved clinical remission at week 16. There were no safety concerns. Non-invasive ta-VNS reduced FC levels and improved symptoms in a pediatric cohort with mild/moderate IBD. Further research is needed to identify optimal electrical dose and settings to achieve peak anti-inflammatory effect and to identify the mechanistic principles for this therapy.


Development of a highly sensitive liquid biopsy platform to detect clinically-relevant cancer mutations at low allele fractions in cell-free DNA.
Detection and monitoring of circulating tumor DNA (ctDNA) is rapidly becoming a diagnostic, prognostic and predictive tool in cancer patient care. A growing number of gene targets have been identified as diagnostic or actionable, requiring the development of reliable technology that provides analysis of multiple genes in parallel. We have developed the InVision™ liquid biopsy platform which utilizes enhanced TAm-Seq™ (eTAm-Seq™) technology, an amplicon-based next generation sequencing method for the identification of clinically-relevant somatic alterations at low frequency in ctDNA across a panel of 35 cancer-related genes. We present analytical validation of the eTAm-Seq technology across two laboratories to determine the reproducibility of mutation identification. We assess the quantitative performance of eTAm-Seq technology for analysis of single nucleotide variants in clinically-relevant genes as compared to digital PCR (dPCR), using both established DNA standards and novel full-process control material. The assay detected mutant alleles down to 0.02% AF, with high per-base specificity of 99.9997%. Across two laboratories, analysis of samples with optimal amount of DNA detected 94% mutations at 0.25%-0.33% allele fraction (AF), with 90% of mutations detected for samples with lower amounts of input DNA. These studies demonstrate that eTAm-Seq technology is a robust and reproducible technology for the identification and quantification of somatic mutations in circulating tumor DNA, and support its use in clinical applications for precision medicine.


The endocannabinoid system and nondrug rewarding behaviours.
Rewarding behaviours such as sexual activity, eating, nursing, parenting, social interactions, and play activity are conserved strongly in evolution, and they are essential for development and survival. All of these behaviours are enjoyable and represent pleasant experiences with a high reward value. Remarkably, rewarding behaviours activate the same brain circuits that mediate the positive reinforcing effects of drugs of abuse and of other forms of addiction, such as gambling and food addiction. Given the involvement of the endocannabinoid system in a variety of physiological functions of the nervous system, it is not surprising that it takes part in the complex machinery that regulates gratification and perception of pleasure. In this review, we focus first on the role of the endocannabinoid system in the modulation of neural activity and synaptic functions in brain regions that are involved in natural and nonnatural rewards (namely, the ventral tegmental area, striatum, amygdala, and prefrontal cortex). Then, we examine the role of the endocannabinoid system in modulating behaviours that directly or indirectly activate these brain reward pathways. More specifically, current knowledge of the effects of the pharmacological manipulation of the endocannabinoid system on natural (eating, sexual behaviour, parenting, and social play) and pathological (gambling) rewarding behaviours is summarised and discussed.


Psychometric Properties of a Measure Assessing Anxiogenic Parenting Practices in Food Allergy.
Pediatric food allergy represents a significant public health burden. In order to avoid allergen consumption, adequate management requires daily vigilance and involvement from parents, frequently leading to increased parental anxiety. While specific anxiogenic parenting practices (i.e., parenting behaviors which may aid in the development and/or exacerbation of childhood anxiety) have been documented within this population, to this point, these behaviors have not been systematically measured. The current study aimed to develop and examine a parent-report scale designed to measure anxiogenic parenting behaviors related to food allergy. Participants included 177 parents of children with food allergy recruited online using Amazon Mechanical Turk (mTurk). An exploratory factor analysis was conducted to determine the factor structure of the newly developed scale. Subsequently, psychometric properties (e.g., construct validity) were examined via correlational analyses. Results indicated a 24-item, 3 factor (Factor 1: Involvement in Food Allergy; Factor 2: Worry about Food Allergy; Factor 3: Autonomy Promotion) scale, which accounted for 53.11% of the total variance. The Kaiser-Meyer-Olkin measure was acceptable, KMO = 0.872 and Bartlett's test of sphericity indicated sufficient correlations between items (χ2(378)=2568.95). All subscales demonstrated strong internal consistency (Involvement in Food Allergy: α = .880; Worry about Food Allergy: α = .892; Autonomy Promotion α = .796) as well as convergent and discriminant validity. Results support the overall psychometric properties of the scale. Interpretations, limitations, and future directions are discussed.


Battery Charge Affects the Stability of Light Intensity from Light-emitting Diode Light-curing Units.
This study investigated the influence of battery charge levels on the stability of light-emitting diode (LED) curing-light intensity by measuring the intensity from fully charged through fully discharged batteries. The microhardness of resin composites polymerized by the light-curing units at various battery charge levels was measured. The light intensities of seven fully charged battery LED light-curing units-1) LY-A180, 2) Bluephase, 3) Woodpecker, 4) Demi Plus, 5) Saab II, 6) Elipar S10, and 7) MiniLED-were measured with a radiometer (Kerr) after every 10 uses (20 seconds per use) until the battery was discharged. Ten 2-mm-thick cylindrical specimens of A3 shade nanofilled resin composite (PREMISE, Kerr) were prepared per LED light-curing unit group. Each specimen was irradiated by the fully charged light-curing unit for 20 seconds. The LED light-curing units were then used until the battery charge fell to 50%. Specimens were prepared again as described above. This was repeated again when the light-curing units' battery charge fell to 25% and when the light intensity had decreased to 400 mW/cm2. The top/bottom surface Knoop hardness ratios of the specimens were determined. The microhardness data were analyzed by one-way analysis of variance with Tukey test at a significance level of 0.05. The Pearson correlation coefficient was used to determine significant correlations between surface hardness and light intensity. We found that the light intensities of the Bluephase, Demi Plus, and Elipar S10 units were stable. The intensity of the MiniLED unit decreased slightly; however, it remained above 400 mW/cm2. In contrast, the intensities of the LY-A180, Woodpecker, and Saab II units decreased below 400 mW/cm2. There was also a significant decrease in the surface microhardnesses of the resin composite specimens treated with MiniLED, LY-A180, Woodpecker, and Saab II. In conclusion, the light intensity of several LED light-curing units decreased as the battery was discharged, with a coincident reduction in the units' ability to polymerize resin composite. Therefore, the intensity of an LED light-curing unit should be evaluated during the life of its battery charge to ensure that sufficient light intensity is being generated.


Adjuvant radiation for salivary gland malignancies is associated with improved survival: A National Cancer Database analysis.
There are no randomized data to support the use of postoperative radiation for salivary gland malignancies. This study uses the National Cancer Database (NCDB) to describe the epidemiology of salivary gland cancer patients and to investigate whether treatment with adjuvant radiation improves overall survival. A total of 8243 patients diagnosed with a major salivary gland cancer were identified from the NCDB. All patients received primary surgical resection of their malignancy. Patients were risk-stratified by adverse features, and overall survival rates were determined. Patients were considered high risk if they had extracapsular extension and/or positive margin after resection. Patients were considered intermediate risk if they did not meet the criteria for high risk but had pT3-T4 disease, pN+ disease, lymphovascular space invasion, adenoid cystic histology, or grade 2-3 disease. Patients who did not meet criteria for high or intermediate risk were considered low risk. Overall patient demographics, disease characteristics, treatment factors, and outcomes were summarized with descriptive statistics and analyzed with STATA. Median follow-up in this cohort was 42.4 months, with the median age of 58 years. Patients in the high-risk group had greater survival (hazard ratio [HR], 0.76; P = .002; 95% confidence interval [CI], 0.64-0.91) if they received adjuvant radiation therapy. In contrast, patients in the intermediate- (HR, 1.01; P = .904; 95% CI, 0.85-1.20) and low-risk groups (HR, 0.85; P = .427; 95% CI, 0.57-1.26) did not experience a survival benefit with adjuvant radiation therapy. This large analysis compared survival outcomes between observation and adjuvant radiation alone in risk-stratified patients after resection of major salivary glands using a national database. The use of adjuvant radiation for high-risk major salivary gland cancers appears to offer a survival benefit. Although an overall survival benefit was not seen in low- and intermediate-risk salivary gland cancers, this study could not address impact on local control because of the limitations of the NCDB.


Transcriptional Regulation of Energy Metabolism in Cancer Cells.
Cancer development, growth, and metastasis are highly regulated by several transcription regulators (TRs), namely transcription factors, oncogenes, tumor-suppressor genes, and protein kinases. Although TR roles in these events have been well characterized, their functions in regulating other important cancer cell processes, such as metabolism, have not been systematically examined. In this review, we describe, analyze, and strive to reconstruct the regulatory networks of several TRs acting in the energy metabolism pathways, glycolysis (and its main branching reactions), and oxidative phosphorylation of nonmetastatic and metastatic cancer cells. Moreover, we propose which possible gene targets might allow these TRs to facilitate the modulation of each energy metabolism pathway, depending on the tumor microenvironment.


Comorbidity of Lifetime Alcohol Use Disorder and Major Depressive Disorder in San Juan, Puerto Rico.
The purpose of this study was to update estimates of comorbidity between lifetime alcohol use disorder (AUD) severity and lifetime major depressive disorder (MDD) in San Juan, Puerto Rico. Data are from a household random sample of 1,510 individuals (816 female) 18-64 years of age in San Juan, Puerto Rico. AUD and MDD identification follow criteria from the Diagnostic and Statistical Manual of Mental Disorders, Fifth Edition and Fourth Edition, respectively, both implemented with the Composite International Diagnostic Interview (CIDI). It is possible to implement DSM-5 AUD identification with the CIDI, but only DSM-IV criteria can be applied to identify MDD. The prevalence of lifetime MDD was 11% among men, 17% among women, and 14% for both genders; the prevalence of lifetime AUD was 38% among men, 16% among women, and 26% for both genders. Among those with AUD, the rate of MDD was 17% among men and 35% among women. Among those without AUD the rate of MDD was 7% among men and 15% among women. Results of multiple logistic regression analysis controlling for gender, illegal drug use, age, level of family cohesion, religion, employment status, marital status, education, and family annual income showed that AUD severity was positively associated with the likelihood of MDD, as follows: mild AUD, adjusted odds ratio [AOR] = 1.78 (95% CI [1.09, 2.91], p < .05); moderate AUD, AOR = 2.58 (95% CI [1.33, 5.01], p < .01); and severe AUD, AOR = 3.34 (95% CI [1.70, 6.56], p < .01). MDD frequently occurs as a comorbid condition with AUD in San Juan, Puerto Rico. The frequency of occurrence increases as AUD severity increases. AUD treatment providers should therefore be equally prepared to treat these two comorbid conditions.


Palladium-Catalyzed Three-Component Coupling of Ynamides.
A palladium-catalyzed regioselective three-component coupling of ynamides was developed. The reaction proceeded smoothly to furnish the desired products when carried out at 70 °C in acetonitrile/water with potassium carbonate in the presence of 2.5 mol % Pd2(dba)3·CHCl3 without a ligand. Various iodides and boronic acids were used in this reaction, and a carbon-carbon bond was formed with satisfactory regioselectivity from the ynamides.


Defining the scope for altering rice leaf anatomy to improve photosynthesis: a modelling approach.
Leaf structure plays an important role in photosynthesis. However, the causal relationship and the quantitative importance of any single structural parameter to the overall photosynthetic performance of a leaf remains open to debate. In this paper, we report on a mechanistic model, eLeaf, which successfully captures rice leaf photosynthetic performance under varying environmental conditions of light and CO2 . We developed a 3D reaction-diffusion model for leaf photosynthesis parameterised using a range of imaging data and biochemical measurements from plants grown under ambient and elevated CO2 and then interrogated the model to quantify the importance of these elements. The model successfully captured leaf-level photosynthetic performance in rice. Photosynthetic metabolism underpinned the majority of the increased carbon assimilation rate observed under elevated CO2 levels, with a range of structural elements making positive and negative contributions. Mesophyll porosity could be varied without any major outcome on photosynthetic performance, providing a theoretical underpinning for experimental data. eLeaf allows quantitative analysis of the influence of morphological and biochemical properties on leaf photosynthesis. The analysis highlights a degree of leaf structural plasticity with respect to photosynthesis of significance in the context of attempts to improve crop photosynthesis.


[Cardiovascular risk in systemic inflammatory diseases].
Systemic inflammatory diseases are associated with increased cardiovascular morbidity and mortality. The link between inflammatory and cardiovascular diseases can be attributed to the coexistence of classical risk factors and inflammatory mechanisms activated during systemic inflammatory diseases involving the immune system. Unfavorable metabolic effects of anti-inflammatory drugs can also contribute to increase cardiovascular risk. Yet, clinical implications of these findings are not entirely clear, and deeper knowledge and awareness of cardiac involvement in inflammatory diseases are necessary. The aim of this review is to summarize cardiac involvement in systemic inflammatory diseases and to identify aspects where evidence is currently lacking that would deserve further investigation in the future.


Emerging Trends in Pain Modulation by Metabotropic Glutamate Receptors.
Pain is an essential protective mechanism meant to prevent tissue damages in organisms. On the other hand, chronic or persistent pain caused, for example, by inflammation or nerve injury is long lasting and responsible for long-term disability in patients. Therefore, chronic pain and its management represents a major public health problem. Hence, it is critical to better understand chronic pain molecular mechanisms to develop innovative and efficient drugs. Over the past decades, accumulating evidence has demonstrated a pivotal role of glutamate in pain sensation and transmission, supporting glutamate receptors as promising potential targets for pain relieving drug development. Glutamate is the most abundant excitatory neurotransmitter in the brain. Once released into the synapse, glutamate acts through ionotropic glutamate receptors (iGluRs), which are ligand-gated ion channels triggering fast excitatory neurotransmission, and metabotropic glutamate receptors (mGluRs), which are G protein-coupled receptors modulating synaptic transmission. Eight mGluRs subtypes have been identified and are divided into three classes based on their sequence similarities and their pharmacological and biochemical properties. Of note, all mGluR subtypes (except mGlu6 receptor) are expressed within the nociceptive pathways where they modulate pain transmission. This review will address the role of mGluRs in acute and persistent pain processing and emerging pharmacotherapies for pain management.


Autologous hematopoietic stem cell transplantation modifies specific aspects of systemic sclerosis-related microvasculopathy.
Autologous hematopoietic stem cell transplantation (AHSCT) is a therapeutic option for patients with severe and progressive systemic sclerosis (SSc). Here, we aimed to investigate how AHSCT affects the vasculopathy of SSc patients. Twenty-seven SSc patients were retrospectively assessed, before and after AHSCT, for vessel morphology (nailfold capillaroscopy), skin expression of endothelial markers and serum levels of markers of inflammation, angiogenesis and endothelial activation. Skin biopsies were analyzed by immunohistochemistry (IHC) for expression of CD31, VE-cadherin, E-selectin, angiopoietin-1 (Ang1), angiopoietin-2 (Ang2), Tie-2, vascular endothelial growth factor A (VEGFA), vascular endothelial growth factor receptor 2 (VEGFR2), and endothelin-1 before and 12 months post-AHSCT. Serum samples from SSc patients were assessed before and up to 36 months after AHSCT for IL-6, von Willebrand factor (vWF), CXC Motif Chemokine Ligand 8 (CXCL8), Endothelin-1, epidermal growth factor (EGF), VEGFA, Pentraxin-3, Intercellular Adhesion Molecule 1 (ICAM-1), E-selectin, P-selectin, Thrombomodulin and IL-18 levels, and compared to healthy control samples. On nailfold capillaroscopy, the number of capillaries increased at 1 year, while giant capillaries decreased at 6 months and 1 year after AHSCT. In the skin biopsies, expression of E-selectin notably decreased and Ang1 increased after AHSCT. At baseline, all vascular markers evaluated in the serum were significantly higher in SSc patients when compared to healthy controls, except for ICAM-1. When compared at different time points after AHSCT, Thrombomodulin, Pentraxin-3, vWF, and IL-18 levels remained generally stable at high levels until 36 months after AHSCT. Our results suggest that AHSCT contributes to improvements of the vessel morphology and dermal microvasculopathy, but does not normalize elevated levels of serum vascular markers in SSc patients. Additional vascular therapeutic approaches might contribute to more effectively treat the endothelial injury.


Description of a new yeast species, Malassezia japonica, and its detection in patients with atopic dermatitis and healthy subjects.
Lipophilic yeasts of the genus Malassezia are part of the normal cutaneous microflora and are considered one of the factors that trigger atopic dermatitis (AD). We isolated two strains of Malassezia from a healthy Japanese female. Analysis of the D1/D2 26S ribosomal DNA and internal transcribed spacer region sequences of the isolates suggested that they are new members of the genus Malassezia. We propose the name Malassezia japonica sp. nov. for the isolates. M. japonica is easily distinguished from the seven known lipophilic species by its ability to assimilate Tween 40 and Tween 60 and its inability to assimilate Tween 20 and Tween 80 and to grow at 40 degrees C. Furthermore, by applying transparent dressings to the skin lesions of 36 patients with AD and the skin of 22 healthy subjects, M. japonica DNA was detected by a non-culture-based method consisting of nested PCR with M. japonica species-specific primers. M. japonica DNA was detected from 12 of the 36 patients (33.3%) and 3 of the 22 healthy subjects (13.6%). Although it is not known whether M. japonica plays a role in AD, this species was part of the microflora in both patients with AD and healthy subjects.


Proliferation, migration, and neuronal differentiation of the endogenous neural progenitors in hippocampus after fimbria fornix transection.
Neurogenesis in the hippocampus continues throughout adult life and can be regulated by the local microenvironment. To determine whether denervation stimulates neurogenesis in hippocampus, proliferation, migration, and differentiation of local neural stem cells (NSCs) in dentate gyrus was investigated after fimbria fornix transection. In the denervated hippocampus, NSCs proliferated markedly and migrated along the subgranular layer, and more newborn cells differentiated into neurons or astrocytes. After denervation, more newborn cells in the deafferented hippocampus expressed Brn-4 and differentiated into beta-Tubulin III positive neurons. It is concluded that the local NSCs in hippocampus may proliferate and migrate into granule cell layer, in which changes in the deafferented hippocampus provided a suitable microenvironment for hippocampal neurogenesis and the increased Brn-4 in denervated hippocampus may be involved in this process.


Pre-B cell colony enhancing factor induces Nampt-dependent translocation of the insulin receptor out of lipid microdomains in A549 lung epithelial cells.
Pre-B cell colony-enhancing factor (PBEF) is a highly conserved pleiotropic protein reported to be an alternate ligand for the insulin receptor (IR). We sought to clarify the relationship between PBEF and insulin signaling by evaluating the effects of PBEF on the localization of the IRβ chain to lipid rafts in A549 epithelial cells. We isolated lipid rafts from A549 cells and detected the IR by immunoprecipitation from raft fractions or whole cell lysates. Cells were treated with rPBEF, its enzymatic product nicotinamide adenine dinucleotide (NAD), or the Nampt inhibitor daporinad to study the effect of PBEF on IRβ movement. We used coimmunoprecipitation studies in cells transfected with PBEF and IRβ constructs to detect interactions between PBEF, the IRβ, and caveolin-1 (Cav-1). PBEF was present in both lipid raft and nonraft fractions, whereas the IR was found only in lipid raft fractions of resting A549 cells. The IR-, PBEF-, and Cav-1-coimmunoprecipitated rPBEF treatment resulted in the movement of IRβ- and tyrosine-phosphorylated Cav-1 from lipid rafts to nonrafts, an effect that could be blocked by daporinad, suggesting that this effect was facilitated by the Nampt activity of PBEF. The addition of PBEF to insulin-treated cells resulted in reduced Akt phosphorylation of both Ser⁴⁷³ and Thr³⁰⁸. We conclude that PBEF can inhibit insulin signaling through the IR by Nampt-dependent promotion of IR translocation into the nonraft domains of A549 epithelial cells. PBEF-induced alterations in the spatial geometry of the IR provide a mechanistic explanation for insulin resistance in inflammatory states associated with upregulation of PBEF.


Quantitative binding of antibiotics to ribosomes from a yeast mutant altered on the peptidyl-transferase center.
Quantitative binding studies of [G-3H]anisomycin and [acetyl-14C]trichodermin to sensitive and resistant 80-S ribosomes from yeasts are described in this work. A single mutation, most probably affecting the ribosome peptidyl transferase centre, appears to have pleiotropic effects on the ribosome leading to resistance to trichodermin and anisomycin and to an increased sensitivity to sparsomycin. Resistance to trichodermin is due to a reduced affinity of ribosomes from the mutant for the antibiotic. Ribosomes from the sensitive strain (Y 1661 bind [acetyl-14C]trichodermin with a dissociation constant of 0.99 muM while those from the resistant one (TR1) bind [acetyl-14C]trichodermin with a dissociation constant of 15.4 muM. Similar results are obtained when the binding of [acetyl-14C]trichodermin to Y 166 and TR1 60-S subunits is studied. The mutant TR1 is also resistant to anisomycin. Although trichodermin and anisomycin bind to the ribosome at mutually exclusive sites, the higher affinity binding of [G-3H]anisomycin that is responsible for the inhibition of the peptidyl transferase center is practically identical for Y 166 and TR1 ribosomes. Therefore, the mutation in the ribosome leading to resistance to trichodermin and anisomycin decreases the affinity for trichodermin but not for anisomycin. Trichodermin, trichothecin and fusarenon X inhibit the binding of [G-3H]anisomycin to TR1 ribosomes to a lower extent than to Y 166 ribosomes, suggesting that the resistance of TR1 ribosomes to the effects of trichothecin and fusarenon X is caused by a decrease in the affinity of the ribosomes for these drugs, as was seen with trichodermin. On the other hand, verrucarin A inhibits [G-3H]anisomycin binding to Y 166 and TR1 ribosomes to a similar extent and therefore its affinity for the ribosome does not appear to be affected by the mutation leading to resistance. Trichothecin, trichodermin and fusarenon X appear to have a common binding site on the 60-S ribosomal subunits, which overlaps or is closely linked to the binding sites of anisomycin and verrucarin A.


Improved osteogenic differentiation of human induced pluripotent stem cells cultured on polyvinylidene fluoride/collagen/platelet-rich plasma composite nanofibers.
Blood transfusion or blood products, such as plasma, have a long history in improving health, but today, platelet-rich plasma (PRP) is used in various medical areas such as surgery, orthopedics, and rheumatology in many ways. Considering the high efficiency of tissue engineering in repairing bone defects, in this study, we investigated the combined effect of nanofibrous scaffolds in combination with PRP on the osteogenic differentiation potential of human induced pluripotent stem cells (iPSCs). Electrospinning was used for fabricating nanofibrous scaffolds by polyvinylidene fluoride/collagen (PVDF/col) with and without PRP. After scaffold characterization, the osteoinductivity of the fabricated scaffolds was studied by culturing human iPSCs under osteogenic medium. The results showed that PRP has a considerable positive effect on the biocompatibility of the PVDF/col nanofibrous scaffold when examined by protein adsorption, cell attachment, and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assays. In addition, the results obtained from alkaline phosphatase activity and calcium content assays demonstrated that nanofibers have higher osteoinductivity while grown on PRP-incorporated PVDF/col nanofibers. These results were also confirmed while the osteogenic differentiation of the iPSCs was more investigated by evaluating the most important bone-related genes expression level. According to the results, it can be concluded that PVDF/col/PRP has much more osteoinductivity while compared with the PVDF/col, and it can be introduced as a promising bone bio-implant for use in bone tissue engineering applications.


UC/MALDI-MS analysis of HDL; evidence for density-dependent post-translational modifications.
The purpose of this study is to determine whether the nature of the post-translational modifications of the major apolipoproteins of HDL is different for density-distinct subclasses. These subclasses were separated by ultracentrifugation using a novel density-forming solute to yield a high-resolution separation. The serum of two subjects, a control with a normolipidemic profile and a subject with diagnosed cardiovascular disease, was studied. Aliquots of three HDL subclasses were analyzed by MALDI and considerable differences were seen when comparing density-distinct subclasses and also when comparing the two subjects. A detailed analysis of the post-translational modification pattern of apoA-1 shows evidence of considerable protease activity, particularly in the more dense fractions. We conclude that part of the heterogeneity of the population of HDL particles is due to density-dependent protease activity.


Apatinib-loaded nanoparticles suppress vascular endothelial growth factor-induced angiogenesis and experimental corneal neovascularization.
Pathological angiogenesis is one of the major symptoms of severe ocular diseases, including corneal neovascularization. The blockade of vascular endothelial growth factor (VEGF) action has been recognized as an efficient strategy for treating corneal neovascularization. In this study, we aimed to investigate whether nanoparticle-based delivery of apatinib, a novel and selective inhibitor of VEGF receptor 2, inhibits VEGF-mediated angiogenesis and suppresses experimental corneal neovascularization. Water-insoluble apatinib was encapsulated in nanoparticles composed of human serum albumin (HSA)-conjugated polyethylene glycol (PEG). In vitro angiogenesis assays showed that apatinib-loaded HSA-PEG (Apa-HSA-PEG) nanoparticles potently inhibited VEGF-induced tube formation, scratch wounding migration, and proliferation of human endothelial cells. In a rat model of alkali burn injury-induced corneal neovascularization, a subconjunctival injection of Apa-HSA-PEG nanoparticles induced a significant decrease in neovascularization compared to that observed with an injection of free apatinib solution or phosphate-buffered saline. An in vivo distribution study using HSA-PEG nanoparticles loaded with fluorescent hydrophobic model drugs revealed the presence of a substantial number of nanoparticles in the corneal stroma within 24 h after injection. These in vitro and in vivo results demonstrate that apatinib-loaded nanoparticles may be promising for the prevention and treatment of corneal neovascularization-related ocular disorders.


Is there a connection between gestational diabetes mellitus, human immunodeficiency virus infection, and tuberculosis?
Pregnancy is associated with insulin resistance similar to that found in type 2 diabetes mellitus (DM). The prevalence of gestational diabetes mellitus (GDM) in key tuberculosis (TB) endemic countries, such as India and China, has been increasing rapidly in the last decade and may be higher in human immunodeficiency virus (HIV) infected women. Pregnancy is also an independent risk factor for developing active TB; however, little is known about the interaction of GDM, HIV and TB. We review the epidemiology and immunology of GDM, and significant research gaps in understanding the interactions between GDM, pregnancy, and TB in women living with and those without HIV.


Adsorption of Homotrifunctional 1,2,3-Benzenetriol on a Ge(100)-2 × 1 Surface.
The adsorption of the homotrifunctional 1,2,3-benzenetriol on Ge(100)-2 × 1 has been investigated by density functional theory calculations, Fourier transform infrared spectroscopy, and X-ray-photoelectron spectroscopy. The results show that the adsorption can occur through OH dissociation of all three hydroxyl groups, and that all three reaction pathways are kinetically and thermodynamically favorable. A coverage-dependent analysis shows that at low coverage, the molecule reacts to form a mix of trifold and dually bound adsorbates. As the coverage increases, the reactions are limited to dissociative adsorption through single and dual attachments. Calculations on the three possible dually bound configurations further reveals that the dissociative adsorption of the third hydroxyl group is limited by geometrical constraints to only two reaction channels. Finally, the proximity between OH-groups in the molecule favors intra- and intermolecular hydrogen bonding, which stabilizes singly and dually bound adsorbate configurations and limits the reactivity of the functional groups.


Femtosecond studies: observing transition states and other ultra-short-lived species.
To celebrate 2015 as the 'International Year of Light', this article offers a short survey of the progress made since the award of the Nobel Prize of 1999 to Professor Ahmed Zewail for his pioneering work on taking the timescale for observation of light-induced events down to the femtosecond level. Developments have included the extension of studies (i) to larger molecules, leading up to biological systems; (ii) the increased range of detection methods of transient species from the UV-Vis to the infrared region; (iii) the introduction of Raman spectroscopy to augment IR studies; (iv) examination of combination events to supplement dissociation events; (v) the interrogation of transient structures by X-ray absorption spectroscopy; (vi) the study of reactions taking place at solid surfaces.


Induction of labor using high-dose or low-dose prostaglandin vaginal pessaries.
Two hundred women were studied in a randomized controlled trial of induction of labor using high-dose (3 mg) or low-dose (0.5 mg) prostaglandin E2 vaginal pessaries. Induction using 3-mg pessaries was more successful, with a shorter overall induction-delivery interval and less requirement for analgesia, although uterine hyperstimulation occurred in two patients. In contrast, the low-dose regimen did not cause hypertonus and, though less successful in inducing labor, the total dose of prostaglandin E2 used was considerably less than in the 3-mg group. The cesarean section rate when the cervix was initially unfavorable was high in both groups (high dose 18.2%, low dose 16.7%). The response to prostaglandin E2 did not relate closely to the initial cervical state.


Derivation of cochlea hair cell for in vitro expansion and characterization.
A potential cure for hearing loss would be to regenerate hair cells by stimulating cells of the damaged inner ear sensory epithelia to proliferate and differentiate into hair cells. Here, we investigated the possibility to isolate, culture-expand and characterize the cells from the cochlea membrane of adult mice. Our results showed that the cultured cells isolated from mouse cochlea membrane were heterogenous in nature. Morphologically there were epithelial like cells, hair cell like, nerve cell like and fibroblastic cells observed in the culture. The cultured cells were immunopositive for specific hair cell markers including Myosin 7a, Calretinin and Espin.


[Advances in the microbial production of the compatible solute ectoine: a review].
Ectoine is an amino acid derivative and an important natural product in halophilic microorganisms. It plays an important role in protecting cells and stabilizing biological macromolecules, and can be widely used in biomedical fields such as drug preparation adjuvants, organ transplantation and preservation, skin wound repair and cosmetics. Due to the medical value and commercial market demand of ectoine, this article summarized the recent advances in the microbial production of ectoine, including the mutation and breeding of hyper-producing strains, construction of genetically and metabolically engineered strains, optimization of fermentation processes, and extraction and purification processes. The application of multi-omics technologies and computational biology to develop an ectoine producing cell factory was prospected, with the aim to provide a reference for ectoine overproduction.


Explaining gender differences in jurors' reactions to child sexual assault cases.
In three experiments, we investigated the influence of juror, victim, and case factors on mock jurors' decisions in several types of child sexual assault cases (incest, day care, stranger abduction, and teacher-perpetrated abuse). We also validated and tested the ability of several scales measuring empathy for child victims, children's believability, and opposition to adult/child sex, to mediate the effect of jurors' gender on case judgments. Supporting a theoretical model derived from research on the perceived credibility of adult rape victims, women compared to men were more empathic toward child victims, more opposed to adult/child sex, more pro-women, and more inclined to believe children generally. In turn, women (versus men) made more pro-victim judgments in hypothetical abuse cases; that is, attitudes and empathy generally mediated this juror gender effect that is pervasive in this literature. The experiments also revealed that strength of case evidence is a powerful factor in determining judgments, and that teen victims (14 years old) are blamed more for sexual abuse than are younger children (5 years old), but that perceptions of 5 and 10 year olds are largely similar. Our last experiment illustrated that our findings of mediation generalize to a community member sample.


LncRNA AC136007.2 alleviates cerebral ischemic-reperfusion injury by suppressing autophagy.
Differential expression and diagnostic significance of the long noncoding RNA (lncRNA) AC136007.2 has been reported in patients with acute ischemic stroke (AIS). However, its role on disease progression and outcome remains unclear. Here, we employed an oxygen-glucose deprivation/reperfusion (OGD/R) model in neuronal SH-SY5Y cells and performed middle cerebral artery occlusion (MCAO) surgery in rats to investigate the function of AC136007.2 in ischemia-reperfusion (I/R) injury. AC136007.2 expression was determined by RT-qPCR and cell viability was examined using CCK-8, Edu, LDH, and apoptosis assays. Pro-inflammatory cytokine expression was assessed using ELISA. OGD/R downregulated AC136007.2 expression in SH-SY5Y cells, decreased viability by inducing apoptosis, and stimulated secretion of TNF-α, IL-6, and IL-1β. In turn, lentivirus-mediated AC136007.2 overexpression significantly reversed these phenomena. LC3 immunofluorescence and western blotting analyses of LC3-I/II and Beclin-1 expression and AMPK/mTOR phosphorylation status showed that AC136007.2 suppressed autophagy in SH-SY5Y cells via inactivation of AMPK/mTOR signaling. Notably, incubation with the AMPK activator AICAR abolished the pro-survival effect of AC136007.2 upon OGD/R treatment. Importantly, intraventricular injection of AC136007.2 significantly reduced cerebral infarction and brain edema in MCAO rats, as shown by TTC staining and water content measurements. We conclude that AC136007.2 alleviates cerebral I/R injury by suppressing AMPK/mTOR-dependent autophagy.


Removal and replacement of a fractured dental implant: case report.
To achieve satisfactory function and esthetics following the fracture of a dental implant, the clinician is often faced with the imposing prospect of fixture removal and the restoration of adequate osseous support for subsequent fixture placement and eventual restoration. This article describes a technique that allowed removal of the fractured fixture, conservative elevation of the sinus floor and the immediate insertion of a larger diameter fixture, to permit eventual replacement.


Desmopressin responsiveness in children with Ehlers-Danlos syndrome associated bleeding symptoms.
Ehlers-Danlos Syndrome (EDS) is caused by heritable collagen defects and may be associated with bleeding symptoms. Desmopressin has been described in case reports to decrease bleeding times in these patients. This study sought to assess bleeding time responsiveness to desmopressin therapy in a cohort of children with EDS-associated bleeding manifestations. A retrospective chart review of children with EDS referred for bleeding symptoms was utilized. Twenty-six children were included; 19 (73%) had a desmopressin challenge. The mean bleeding time was 11.26 (+/-4.39) min, decreasing to 5.95 (+/-2.41) min with treatment (P < 0.01). Desmopressin normalizes bleeding times in children with EDS.


[Musculoplasty of the peroneus brevis in ankle joint injuries].
Six patients with ankle joint's injuries had with muscle transposition by peroneus brevis. We observed them for three months. The important point was functional state of sort tissue. Wound healing of these patients were per primam except one of them. None of the patients had functional deficit.


Leader sequences of Strongylocentrotus purpuratus histone mRNAs start at a unique heptanucleotide common to all five histone genes.
We have determined the sequence of the untranslated leader nucleotides of all five histone mRNAs from Strongylocentrotus purpuratus by the dideoxy chain termination method. Total polysomal RNA from sea urchin embryos was used as a substrate for cDNA synthesis primed by specific DNA restriction fragments. Each of the primers was derived from the 5'-terminal part of the coding region for a different histone protein. The five histone mRNA leader sequences are different in length and primary structure. The 5' termini of all five histone mRNAs coincide with the unique heptanucleotide Py-Py-A-T-T-C-Pu in genomic DNA. This sequence, which defines the start of the individual histone mRNAs, is preceded by the A+T-rich octanucleotide identified in front of all eukaryotic structural genes where sequences have been determined to date.


Analysis of curative effect of percutaneous coaxial large channel endoscopic lumbar interbody fusion in the treatment of degenerative lumbar spinal stenosis.
To investigate the clinical efficacy and technical points of Percutaneous Coaxial Large-channel Endoscopic Lumbar Interbody Fusion (PCLE-LIF) in the treatment of degenerative lumbar spinal stenosis. The clinical data of patients with single-segment degenerative lumbar spinal stenosis who underwent PCLE-LIF surgery from January 2019 to June 2021 were retrospectively analyzed. Surgery-related data included symptom duration, operation time, hospital stay, and complication rate. Functional score data included low back pain and lower extremity pain VAS score, ODI score, and MacNab criteria were used to evaluate clinical effects. The Brantigan criteria were used to evaluate the interbody fusion. There were 62 patients in this group, including 35 males and 27 females. The surgical sites were all lower lumbar spine, including 35 cases of lumbar L4/5 and 27 cases of L5/S1. The length of hospital stay was 7.7 ± 1.4 days. All patients were followed up regularly for 1 year. The interbody fusion rate was 93.5% at 1 year after operation. There were 2 cases of numbness, 2 cases of nerve edema and pain, 1 case of cage displacement, and 1 case of pedicle screw loosening. The complication rate was 9.6%. The VAS scores of low back pain 1 day before surgery, 3 days, 3 months and 1 year after surgery were 4.48 ± 1.06, 0.84 ± 0.81, 0.40 ± 0.56, 0.39 ± 0.69, and the VAS of lower extremity pain at each time point of appeal were 5.58 ± 0.98, 0.91 ± 0.58, 0.31 ± 0.46, 0.19 ± 0.40. The ODI scores at 1 day before surgery, 3 months and 1 year after surgery were 60.01 ± 6.21, 15.58 ± 2.84, 8.82 ± 2.15. The ODI scores and VAS scores of low back pain and lower extremity pain at each follow-up time point after operation were significantly lower than those before operation (p < 0.05). The 1-year follow-up after operation was evaluated by the modified MacNab standard, and the results were excellent in 36 cases, good in 23 cases, fair in 3 cases, and poor in 0 cases, with an excellent and good rate of 95.2%. Percutaneous coaxial large-channel endoscopic lumbar interbody fusion in the treatment of degenerative lumbar spinal stenosis has good short-term efficacy and high safety, and is worthy of popularization.


Cell size in Dictyostelium.
Cellular slime mold amoebae have become a model system for the study of cell motility and the cytoskeleton. A basic problem which all cells face that involves the cytoskeleton is how to control their size. The varied ways in which cellular slime mold amoebae change their cell size--by changing the size at which division occurs, by cell fusion, and by control over cytokinesis--are reviewed. A model is presented which attempts to explain how the mechanisms affected in certain cytokinesis mutants in Dictyostelium discoideum known as phg mutants could be involved in control of cell size in the predatory slime mold Dictyostelium caveatum.


[Light-induced retinal damage in pigmented rabbits--1. Histopathological observations of the natural course of healing].
Pigmented rabbits were exposed for 2 hours to xenon light to evaluate the light induced retinal damage clinicopathologically. The xenon light was passed through a heat filter to avoid the influence of long waves, and through the fundus contact lens on the cornea to avoid focusing the light on the retina. No abnormal findings were detected in the retina in fluorescein angiography immediately after the light exposure. Three hours later, hyperfluorescence was found in the center of the exposed area and gradually grew worse. This was most marked twelve hours later. After 6 days, granular hyperfluorescence appeared at the edge of the damaged area and in 2 weeks the whole damaged area showed granular hyperfluorescence. Histopathologically, immediately after the light exposure, slight changes appeared in the retinal pigment epithelium (RPE). Twelve hours later, the cytoplasm of the RPE was edematous, and in 24 hours the RPE became necrotic, but the outer segments of the photoreceptor cells appeared intact. After 6 days, the RPE showed double layering, many macrophages were found in the subretinal space, and the outer segments of the photoreceptor cells were severely damaged. The RPE was primarily damaged and photoreceptor cells were damaged subsequently by xenon light without long waves.


The 19-amino acid cassette of cyclooxygenase-2 mediates entry of the protein into the endoplasmic reticulum-associated degradation system.
Cyclooxygenase (COX) isoforms catalyze the committed step in prostaglandin biosynthesis. The primary structures of COX-1 and COX-2 are very similar except that COX-2 has a 19-amino acid (19-AA) segment of unknown function located just inside its C terminus. Here we provide evidence that the major role of the 19-AA cassette is to mediate entry of COX-2 into the ER-associated degradation system that transports ER proteins to the cytoplasm. COX-1 is constitutively expressed in many cells, whereas COX-2 is usually expressed inducibly and transiently. In murine NIH/3T3 fibroblasts, we find that COX-2 protein is degraded with a half-life (t(1/2)) of about 2 h, whereas COX-1 is reasonably stable (t(1/2) > 12 h); COX-2 degradation is retarded by 26 S proteasome inhibitors. Similarly, COX-1 expressed heterologously in HEK293 cells is quite stable (t(1/2) > 24 h), whereas COX-2 expressed heterologously is degraded with a t(1/2) of approximately 5 h, and its degradation is slowed by proteasome inhibitors. A deletion mutant of COX-2 was prepared lacking 18 residues of the 19-AA cassette. This mutant retains native COX-2 activity but, unlike native COX-2, is stable in HEK293 cells. Conversely, inserting the COX-2 19-AA cassette near the C terminus of COX-1 yields a mutant ins594-612 COX-1 that is unstable (t(1/2) approximately 3 h). Mutation of Asn-594, an N-glycosylation site at the beginning of the 19-AA cassette, stabilizes both COX-2 and ins594-612 COX-1; nonetheless, COX mutants that are glycosylated at Asn-594 but lack the remainder of the 19-amino acid cassette (i.e. del597-612 COX-2 and ins594-596 COX-1) are stable. Thus, although glycosylation of Asn-594 is necessary for COX-2 degradation, at least part of the remainder of the 19-AA insert is also required. Finally, kifunensine, a mannosidase inhibitor that can block entry of ER proteins into the ER-associated degradation system, retards COX-2 degradation.


Analysis of mixed infection of sheep with bluetongue virus serotypes 10 and 17: evidence for genetic reassortment in the vertebrate host.
Two seronegative sheep were infected intravenously with 10(9) PFU each of bluetongue virus (BTV) serotype 10 and BTV serotype 17. One animal experienced a mild bluetongue-like disease, and both experienced a short-duration viremia and developed neutralizing immune responses to both virus serotypes. Progeny virus was isolated from venous blood from each animal by using conditions in which reassortment could not have occurred during isolation. Electropherotypes were determined for the progeny viruses from the infected sheep, yielding strikingly similar results for the two animals. In both sheep, serotype 10 dominated among the progeny, accounting for 92% of the progeny. Serotype 17 was rarely isolated and accounted for 3% of the progeny analyzed. The remaining 5% of the progeny clones were reassortant and derived genome segments from both serotypes 10 and 17. Analysis of the parental origin of genome segments in the small number of reassortant progeny analyzed suggested that selection of specific genome segments may have occurred in the infected sheep. These data indicate that reassortment of genome segments occurs, at low frequency, in sheep mixedly infected with BTV.


[Influence of reagent evaporation on other laboratory reagents and appropriate countermeasures].
When the automated analyzer was first introduced, abnormally high values of urine creatinine and abnormally low values of serum creatinine were seen. It was shown that evaporation of the reducing agent stabilizer(sulfurous acid), which is present in the serum iron measurement kit, was occurring and that the observed low values of serum creatinine were due to consumption of hydrogen peroxide by gasifying sulfurous acid. After the cause was elucidated, the abnormal lowering of creatinine values was prevented. This experience demonstrates that care must be exercised when introducing new analytical method in clinical laboratories because evaporation of reagents can influence in other reagent systems.


Effect of bromo-ergocryptine on serum hPRL, hLH, hFSH, and estradiol 17-beta in women with galactorrhea-amenorrhea.
Fourteen women with amenorrhea-galactorrhea were treated with bromo-ergocryptine. Serum prolactin hPRL, hLH, hFSH, estradiol 17-beta (E2), and progesterone values were determined before and during treatment. No consistent pretreatment hormonal pattern was found. During treatment hPRL levels fell in 13 of 14 patients, and E2 rose in 12 of 14 patients. Levels of hLH became normal, and amounts of hFSH did not change. Galactorrhea lessened in all cases and ceased in 11. Menses resumed in 11 patients and 3 women became pregnant. Cessation of galactorrhea and resumption of menses occurred irrespective of initial hPRL or hLH levels. It is suggested that improvement of galactorrhea and resumption of menses in some patients with normal hPRL values are the result of increased serum E2 levels and may be due to a direct action of bromo-ergocryptine on the ovary. Bromo-ergocryptine may also be effective in treatment of amenorrhea, unassociated with galactorrhea or elevated hPRL levels.


The GTP pool in Bacillus brevis and its significance for sporulation.
The GTP pool of Bacillus brevis as well as that of other nucleotides is highly sensitive to all kinds of environmental changes like the cell transfer procedures or nutrient depletion of the cells. In growing cultures, as well as in cells transferred from rich to nitrogen-deficient medium, the nucleotide pool decreases significantly. This decrease is followed by the onset of sporulation only when cells are allowed to produce the peptide antibiotic tyrocidine or if tyrocidine is added to the culture. However, exogenous tyrocidine is active in triggering sporulation only when it is added within a short period of time immediately after shift down, that is when the nucleotide pool is observed to decrease.


Carbonate apatite versus β-tricalcium phosphate for rat vertical bone augmentation: A comparison of bioresorbable bone substitutes using polytetrafluoroethylene tubes.
This study radiologically and histologically compared two bioresorbable bone substitutes with different compositions carbonate apatite (Cytrans® Granules; CGs) and β-tricalcium phosphate (β-TCP) for vertical bone augmentation on a rat skull using a polytetrafluoroethylene (PTFE) tubes. This PTFE tube was placed at the center of the skull, fixed with Super Bond, and augmented with CGs or β-TCP granules. Specimens with surrounding tissue were harvested at 4, 8, and 12 weeks postoperatively, and radiological and histological evaluations were performed. The bone volume to total volume ratio (BV/TV) of the β-TCP-implanted group was markedly higher than that of the CG-implanted group at 4 and 12 weeks postoperatively. Compared to CGs, β-TCP exhibited the ability to form blood vessels into the graft material for a short period after transplantation, as well as an elevated production of collagen into β-TCP granules during the bone formation process.


First-pass metabolism of ONO-5046 (N-[2-[4-(2,2-dimethylpropionyloxy) phenylsulfonylamino]benzoyl]aminoacetic acid), a novel elastase inhibitor, in rats.
The first-pass metabolism in the intestine and liver of ONO-5046 (N-[2-[4-(2,2-dimethylpropionyloxy)-phenylsulfonylamino]benz oyl]aminoacetic acid), a newly synthesized elastase inhibitor, was separately estimated in rats. When ONO-5046 solution was administered into the whole intestine via the bile duct at a dose of 5 mumol/rat, the extent of bioavailability was only 1.5%. A small but significant increase in the bioavailability with an increase in the dose suggested marked first-pass metabolism with a saturable process. Hepatic first-pass metabolism was estimated by determining the hepatic extraction ratio of ONO-5046 after administration into the portal vein at two different infusion rates (5 mumol/kg/9 min or 5 mumol/kg/20s). The extraction ratio was relatively small and constant (about 20%) under 2 different infusion rates of the drug. Intestinal first-pass metabolism was estimated by determining the drug recovery in the mesenteric plasma after administering the drug into the intestinal loop in situ (mesenteric blood collecting method in situ). The recovery percentage of ONO-5046 in the mesenteric plasma was small (2.58 +/- 0.04% at a dose of 1 mumol/rat), and the remaining ONO-5046 recovered in the mesenteric plasma and in the intestinal loop was a metabolite of ONO-5046 (El-601, N-[2-[(4-hydroxyphenyl)sulfonylamino]benzoyl]amino-acetic acid). Recovery percentage of ONO-5046 in the mesenteric plasma increased significantly with an increase in the dose, although the recovery percentage was still low, even at a higher dose (9.55 +/- 1.17% of dose at a dose of 5 mumol/rat). These results indicate that the low oral bioavailability of ONO-5046 in vivo is mainly due to the marked intestinal first-pass metabolism, including the metabolism in the intestinal fluid, and the dose-dependent oral bioavailability was derived from the saturable intestinal first-pass metabolism.


Procedure for detecting and confirming pentobarbital residues in dog food by gas chromatography/mass spectrometry.
The method described detects and confirms presence of pentobarbital residues in dry, extruded feeds at concentrations of 5-20 ppb. Dried feed is ground to a uniform powder and shaken overnight in methanol. A portion of the methanolic extract is evaporated, and the residue is reconstituted in phosphate-buffered saline. The aqueous extract is cleaned with a solid-phase extraction cartridge designed to extract barbiturate residues from biological matrixes. Dimethyl sulfoxide, tetramethylammonium hydroxide, and iodomethane are added to derivatize pentobarbital, 1,3-Dimethyl-pentobarbital is then acidified with dilute hydrochloric acid and extracted with isooctane. The organic layer is transferred and evaporated under a stream of nitrogen. The residue is reconstituted in a small volume of ethyl acetate for analysis by gas chromatography/mass spectrometry. The limit of detection is approximately 0.7 ppb. The method was validated with pentobarbital-fortified feed samples containing high concentrations of meat and bone meal.


Avian superior olivary nucleus provides divergent inhibitory input to parallel auditory pathways.
The avian auditory brainstem displays parallel processing, a fundamental feature of vertebrate sensory systems. Nuclei specialized for temporal processing are largely separate from those processing other aspects of sound. One possible exception to this parallel organization is the inhibitory input provided by the superior olivary nucleus (SON) to nucleus angularis (NA), nucleus magnocellularis (NM), and nucleus laminaris (NL) and contralateral SON (SONc). We sought to determine whether single SON neurons project to multiple targets or separate neuronal populations project independently to individual target nuclei. We introduced two different fluorescent tracer molecules into pairs of target nuclei and quantified the extent to which retrogradely labeled SON neurons were double labeled. A large proportion of double-labeled SON somata were observed in all cases in which injections were made into any pair of ipsilateral targets (NA and NM, NA and NL, or NM and NL), suggesting that many individual SON neurons project to multiple targets. In contrast, when injections involved the SONc and any or all of the ipsilateral targets, double labeling was rare, suggesting that contralateral and ipsilateral targets are innervated by distinct populations of SON neurons arising largely from regionally segregated areas of SON. Therefore, at the earliest stages of auditory processing, there is interaction between pathways specialized to process temporal cues and those that process other acoustic features. We present a conceptual model that incorporates these results and suggest that SON circuitry, in part, functions to offset interaural intensity differences in interaural time difference processing.


Safety and efficacy of a glucagon-like peptide-1 receptor agonist added to basal insulin therapy versus basal insulin with or without a rapid-acting insulin in patients with type 2 diabetes: results of a meta-analysis.
To consolidate the evidence from randomized controlled trials evaluating the use of glucagon-like peptide-1 receptor agonists (GLP-1 RAs) as add-on to basal insulin therapy in type 2 diabetes (T2D) patients. We searched the EMBASE® and NCBI PubMed (Medline) databases and relevant congress abstracts for randomized controlled trials evaluating the efficacy and safety of GLP-1 RAs as add-on to basal insulin compared with basal insulin with or without rapid-acting insulin (RAI) through 23 May 2016. The pooled data were analyzed using a random-effects meta-analysis model. A subanalysis was performed for trials investigating basal insulin plus GLP-1 RAs versus basal insulin plus RAI. Of the 2617 retrieved records, 19 randomized controlled trials enrolling 7,053 patients with T2D were included. Compared with basal insulin ± RAI, reduction in glycated hemoglobin (HbA1c) from baseline (difference in means: -0.48% [95% confidence interval (CI), -0.67 to -0.30]; p < 0.0001) and weight loss (-2.60 kg [95% CI, -3.32 to -1.89]; p < 0.0001) were significantly greater with basal insulin plus GLP-1 RA. The subanalysis similarly showed significant results for change in HbA1c from baseline and for weight loss, as well as a significantly lower risk of symptomatic hypoglycemia in patients treated with basal insulin plus GLP-1 RA versus basal insulin plus RAI (odds ratio, 0.52 [95% CI, 0.42 to 0.64]; p < 0.0001). Addition of GLP-1 RA to basal insulin provided improved glycemic control, led to weight reduction and similar hypoglycemia rates versus an intensified insulin strategy; however, symptomatic hypoglycemia rates were significantly lower when compared with a basal insulin plus RAI.


Production and fragmentation of multiply charged ions in 'electron-free' matrix-assisted laser desorption/ionization.
An unusually large fraction of multiply charged ions is observed in 'electron-free' matrix-assisted laser desorption/ionization (MALDI). Here we investigate how the yield of multiply charged ions depends on experimental parameters in MALDI. It is found to increase if measures are taken to limit the number of electrons in the plume, for example, by using non-metallic MALDI targets or low laser pulse energies. The ionization energy of the matrix is another important parameter that affects the yield of multiply charged ions: matrices with high ionization energies lead to greater intensities of multiply charged ions. It is furthermore proposed that some of the fragment ions observed in MALDI are due to reactions of analyte with electrons in the plume. The possibility of electron capture dissociation of multiply charged ions produced by MALDI is shown.


Biological markers of acute acrylonitrile intoxication in rats as a function of dose and time.
Three markers of acute acrylonitrile (AN) intoxication, namely, tissue glutathione (GSH), tissue cyanide (CN), and covalent binding to tissue protein, were studied as a function of dose and time. Doses administered and responses expected were 20 mg/kg (LD0), 50 mg/kg (LD10), 80 mg/kg (LD50), and 115 mg/kg (LD90). Liver GSH was the most sensitive marker of AN exposure. At 80 mg/kg AN, virtually complete depletion of liver GSH was observed within 30 min with no recovery through 120 min. Kidney GSH showed a similar, but less intense depletion; while blood and brain GSH were more refractory to AN. Whole blood and brain CN rose progressively during the first 60 min in a dose-dependent fashion. At the lowest dose, CN levels decreased thereafter, whereas, at the three higher doses, CN levels were maintained or continued to increase through 120 min. At the highest dose, blood and brain CN remained at acutely toxic levels through 240 min. Covalent binding increased rapidly in all tissues during the first 30 min at all doses. At the lowest dose, little additional covalent binding was observed beyond 30 min, while at the three higher doses, covalent binding increased, although at a slower rate. The data indicate that these three biologic markers of acute AN intoxication respond dramatically in a time-dependent manner in the toxic dosage range. Furthermore, the data provide evidence that AN toxicity is gated by GSH depletion in liver with the resultant termination of AN detoxification.


Three-dimensional origami paper-based device for portable immunoassay applications.
In this study, we demonstrate a three-dimensional surface-modified origami-paper-based analytical device (3D-soPAD) for immunoassay applications. The platform enables the sequential steps of immunoassays to be easily performed using a folded, sliding paper design featuring multiple pre-stored reagents, allowing us to take advantage of the vertical diffusion of the analyte through the different paper layers. The cellulose substrate is composed of carboxymethyl cellulose modified with N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride and N-hydroxysuccinimide, which provide covalent bonding sites for bio-recognition molecules. After the optimization of the operation parameters, we determined the detection limit of the 3D-soPAD for human immunoglobulin G (HIgG) which can be as low as 0.01 ng mL-1, with a total turnaround time of 7 min. In order to study the long-term storage of the platform, anti-HIgG horseradish peroxidase (aHIgG-HRP) conjugates were stored by freeze-drying in sugar matrices composed of 10% sucrose/10% trehalose (w/w%) on the paper device, retaining 80% of their activity after 75 days of storage at 4 °C. To evaluate the performance of the paper device using real samples, we demonstrated the detection of protein A (a biomarker for Staphylococcus aureus infection) in highly viscous human synovial fluid. These results show that the proposed 3D-soPAD platform can provide sensitive, high-throughput, and on-site prognosis of infection in resource-limited settings.


Determination of Vitamin C, b-carotene and Riboflavin Contents in Five Green Vegetables Organically and Conventionally Grown.
As consumer interest in organically grown vegetables is increasing in Malaysia, there is a need to answer whether the vegetables are more nutritious than those conventionally grown. This study investigates commercially available vegetables grown organically and conventionally, purchased from retailers to analyse β-carotene, vitamin C and riboflavin contents. Five types of green vegetables were selected, namely Chinese mustard (sawi) (Brassica juncea), Chinese kale (kai-lan) (Brassica alboglabra), lettuce (daun salad) (Lactuca sativa), spinach (bayam putih) (Amaranthus viridis) and swamp cabbage (kangkung) (Ipomoea aquatica). For vitamin analysis, a reverse-phase high performance liquid chromatography was used to identify and quantify β -carotene, vitamin C and riboflavin. The findings showed that not all of the organically grown vegetables were higher in vitamins than that conventionally grown. This study found that only swamp cabbage grown organically was highest in β -carotene, vitamin C and riboflavin contents among the entire samples studied. The various nutrients in organically grown vegetables need to be analysed for the generation of a database on nutritional value which is important for future research.


HIV Tat induces a prolonged MYC relocalization next to IGH in circulating B-cells.
With combined antiretroviral therapy (cART), the risk for HIV-infected individuals to develop a non-Hodgkin lymphoma is diminished. However, the incidence of Burkitt lymphoma (BL) remains strikingly elevated. Most BL present a t(8;14) chromosomal translocation which must take place at a time of spatial proximity between the translocation partners. The two partner genes, MYC and IGH, were found colocalized only very rarely in the nuclei of normal peripheral blood B-cells examined using 3D-FISH while circulating B-cells from HIV-infected individuals whose exhibited consistently elevated levels of MYC-IGH colocalization. In vitro, incubating normal B-cells from healthy donors with a transcriptionally active form of the HIV-encoded Tat protein rapidly activated transcription of the nuclease-encoding RAG1 gene. This created DNA damage, including in the MYC gene locus which then moved towards the center of the nucleus where it sustainably colocalized with IGH up to 10-fold more frequently than in controls. In vivo, this could be sufficient to account for the elevated risk of BL-specific chromosomal translocations which would occur following DNA double strand breaks triggered by AID in secondary lymph nodes at the final stage of immunoglobulin gene maturation. New therapeutic attitudes can be envisioned to prevent BL in this high risk group.


A Simple Chamber for Long-term Confocal Imaging of Root and Hypocotyl Development.
Several aspects of plant development, such as lateral root morphogenesis, occur on time spans of several days. To study underlying cellular and subcellular processes, high resolution time-lapse microscopy strategies that preserve physiological conditions are required. Plant tissues must have adequate nutrient and water supply with sustained gaseous exchange but, when submerged and immobilized under a coverslip, they are particularly susceptible to anoxia. One strategy that has been successfully employed is the use of a perfusion system to maintain a constant supply of oxygen and nutrients. However, such arrangements can be complicated, cumbersome, and require specialized equipment. Presented here is an alternative strategy for a simple imaging system using perfluorodecalin as an immersion medium. This system is easy to set up, requires minimal equipment, and is easily mounted on a microscope stage, allowing several imaging chambers to be set up and imaged in parallel. In this system, lateral root growth rates are indistinguishable from growth rates under standard conditions on agar plates for the first two days, and lateral root growth continues at reduced rates for at least another day. Plant tissues are supplied with nutrients via an agar slab that can be used also to administer a range of pharmacological compounds. The system was established to monitor lateral root development but is readily adaptable to image other plant organs such as hypocotyls and primary roots.


The Effect of Parallel-hole Collimator Material on Image and Functional Parameters in SPECT Imaging: A SIMIND Monte Carlo Study.
The collimator in single-photon emission computed tomography (SPECT) is a critical component of the imaging system and plays an impressive role in the imaging quality. In this study, the effect of the collimator material on the radioisotopic image and its functional parameters was studied. The simulating medical imaging nuclear detectors (SIMIND) Monte Carlo program was used to simulate a Siemens E.CAM SPECT (Siemens Medical Solutions, Erlangen, Germany) system equipped with a low-energy high-resolution (LEHR) collimator. The simulation and experimental data from the SPECT imaging modality using (99m)Tc were obtained on a point source and Jaszczak phantom. Seventeen high atomic number materials were considered as LEHR collimator materials. In order to determine the effect of the collimator material on the image and functional parameters, the energy resolution, spatial resolution, contrast, and collimator characteristics parameters such as septal penetration and scatter-to-primary ratio were investigated. Energy spectra profiles, full width at half maximums (FWHMs) (mm) of the point spread function (PSF) curves, system sensitivity, and contrast of cold spheres of the Jaszczak phantom for the simulated and experiment systems have acceptability superimposed. The results of FWHM and energy resolution for the 17 collimators showed that the collimator made of 98% lead and 2% antimony could provide the best FWHM and energy resolution, 7.68 mm and 9.87%, respectively. The LEHR collimator with 98% lead and 2% antimony offers the best resolution and contrast when compared to other high atomic number metals and alloys.


Statistical approaches to harmonize data on cognitive measures in systematic reviews are rarely reported.
To identify statistical methods for harmonization, the procedures aimed at achieving the comparability of previously collected data, which could be used in the context of summary data and individual participant data meta-analysis of cognitive measures. Environmental scan methods were used to conduct two reviews to identify (1) studies that quantitatively combined data on cognition and (2) general literature on statistical methods for data harmonization. Search results were rapidly screened to identify articles of relevance. All 33 meta-analyses combining cognition measures either restricted their analyses to a subset of studies using a common measure or combined standardized effect sizes across studies; none reported their harmonization steps before producing summary effects. In the second scan, three general classes of statistical harmonization models were identified (1) standardization methods, (2) latent variable models, and (3) multiple imputation models; few publications compared methods. Although it is an implicit part of conducting a meta-analysis or pooled analysis, the methods used to assess inferential equivalence of complex constructs are rarely reported or discussed. Progress in this area will be supported by guidelines for the conduct and reporting of the data harmonization and integration and by evaluating and developing statistical approaches to harmonization.


The Covid-19 Pandemic: Limited Water Access and the Precarity of Women Fishers at Lake Kariba, Zimbabwe.
Man-made reservoirs are constructed to meet certain purposes and Lake Kariba, Zimbabwe, was designed for hydroelectric power generation. However, it has developed other multiple uses, and the growth of fisheries on the lake has had a significant impact on the livelihoods of local communities. The declaration of Covid-19 as a pandemic in Zimbabwe in March 2020 was quickly followed by the imposition of national lockdowns with varying levels of severity up to the present day. This was done to curtail the spread of the disease, meanwhile enhancing the nation's capacity in terms of acquiring testing kits, constructing more admission and quarantine centres as well as educating the people about ways to keep safe. In response to the calls by the government to monitor the movement of people and compliance of the lockdown rules, the Zimbabwe National Parks and Wildlife Management Authority (ZNPWMA), the governing body of the Lake Kariba fisheries, imposed rules that have significantly impacted the fishing communities at Lake Kariba. Both gillnet fishers and rod and line fishers have been impacted, but our focus here is on women rod and line fishers. Using the precarious livelihoods conceptual frameworks, we show how the changes in water management during the Covid-19 pandemic lockdowns has generated high levels of precariousness on the livelihoods of women at Kariba. We define precariousness as the condition of uncertainties brought to the women fishers by changes in water restrictions. The precarity of women was induced by several factors. For instance, the women fishers reported that restrictions to accessing fish in areas with high catches impacted them. They are also now obliged to pay exorbitant fishing fees in a way to discourage them to fish; they were frequently chased away from the Lake by ZNPWMA officers; they had limited amount of time to fish due to curfews; and failure to comply results in heavy fines imposed on them among other challenges. We show how these challenges interact with the current Zimbabwe socio-economic crisis to worsen the precariousness and vulnerability of women fishers at Lake Kariba. Data presented in this manuscript are based on participant observation and interviews with women fishers at Lake Kariba.


Informed consent for investigational chemotherapy: patients' and physicians' perceptions.
One hundred forty-four patients and 68 physicians at three cancer centers were studied for their perceptions of the consent procedure, in which they participated one to three weeks earlier, for chemotherapy by one of 65 investigational protocols. Patients recalled the procedure positively and relied heavily on physician's advice. They felt most physicians wanted them to accept; 29% felt their participation in the decision was not encouraged. Primary reasons for accepting were trust in the physician, belief the treatment would help, and fear the disease (viewed as highly serious) would get worse without it. Nearly a fourth did not recall the information given that treatment was investigational. The consent form played no role in decision-making for 69%. Physicians believed therapeutic benefits would exceed potential problems for most patients; they viewed 41% of the patients as less than eager for details of treatment, a third as avoiding the seriousness of the discussion, and a third as passive in decision-making. The perceptual set of both parties places inadvertent constraint on patients' autonomy in decision making.


Splanchnic arterial blood flow in rats with portacaval shunts.
With indirect methods, it was shown in rats with portacaval shunts (PCS) that total hepatic blood flow (THBF) remained constant when expressed per gram of liver. These results implied an absolute increase in hepatic arterial blood flow (HABF). The aim of this study was to investigate HABF and splanchnic nonhepatic arterial blood flow (SNHABF) with a direct method (57Co microspheres) in PCS rats. One month after surgery, the following results were obtained in PCS rats compared with pair-fed, sham-operated rats: 1) liver mass atrophy was 42.3 +/- 10.9%, 2) HABF (ml X min-1 X g liver-1) was increased by a factor of 2.7, 3) SNHABF (ml X min-1 X 100 g body wt-1) was higher (9.8 +/- 3.3 vs. 5.6 +/- 2.7) (P less than 0.05) and 4) THBF (ml X min-1 X g liver-1) was decreased (1.36 +/- 0.34 vs. 1.85 +/- 0.86) but not significantly. Increases in HABF and SNHABF were not the direct consequence of an increase in cardiac output as attested by a normal cerebral blood flow (ml X min-1 X g organ-1) in PCS rats. In PCS rats, an increase in HABF may prevent the further spread of liver necrosis. The cause and the reason for an increase in SNHABF remain unknown.


Survey of California emergency departments about practices for management of suicidal patients and resources available for their care.
To determine the resources available and current practices for the treatment of patients with suicidal ideation or attempts in California emergency departments (EDs). We conducted a mail and e-mail survey of the directors of all 346 EDs in the state of California. Data collected included identification of hospital and respondent, type of hospital, presence of separate psychiatric ED, total number of ED patients and number of ED patients with suicidal ideation or attempts who were treated per week, mental health personnel on call to evaluate suicidal patients, criteria for patient disposition, available disposition options, delays in patient care, changes desired in the ED treatment of suicidal patients, and adequacy of community resources for suicidal patients. Two hundred twenty-three of 346 (64.5%) ED directors responded to the survey. Overall, the mean estimate of the proportion of ED visits by suicidal patients was 1.7%. Though evaluation of patients with suicidal ideation by a mental health professional was the usual practice, 51 respondents (23%) reported that they occasionally send patients with suicidal ideation home without such an evaluation, and 8.5% reported this was done more than 10% of the time. No single type of mental health professional, including psychiatrist, social worker, county or private psychiatric evaluation team, psychiatric nurse, or psychologist, was available for evaluation of suicidal patients in more than 50% of respondent EDs. In the majority of EDs, psychiatric evaluations were performed by either mobile county or private psychiatric evaluation teams or social workers on call to the ED. Psychiatrists were reported to evaluate the majority of suicidal patients in only 10% of EDs. Only 27% of respondents had the ability to admit patients to a psychiatric service at their hospital. When patients needed to be transferred, the estimated mean wait for these transfers was 7 hours. Seventy-one percent of respondents reported needing improved access to mental health personnel for evaluation of suicidal patients; 61% reported needing improved access to mental health personnel for patient disposition. In California EDs, there are limited mental health services for suicidal patients. Regional solutions to emergency and nonemergency mental health problems are needed, including improved access to mental health personnel for ED evaluation, disposition, and follow-up of suicidal patients and community mental health resources for patient referrals.


Diverse patterns of odor representation by neurons in the anterior piriform cortex of awake mice.
The mammalian piriform cortex receives direct synaptic input from the olfactory bulb and is likely the locus for the formation of odor percept. It remains unclear how individual cortical neurons encode olfactory information in unanesthetized animals. By single-cell recordings from head-restrained awake mice, we studied the odor response profiles of individual neurons in the anterior piriform cortex (aPCX). Neurons were juxtacellularly labeled, and their cell types were determined by their morphology and neurotransmitter phenotypes. We found a considerable level of variability in selectivity patterns among pyramidal neurons (PNs). Approximately one-quarter of PNs were broadly activated by structurally dissimilar odorants, whereas the excitations to the rest of PNs were highly selective. Broad inhibition was only observed from a subpopulation of PNs. GABAergic neurons displayed nonselective excitatory responses to test odorants and rarely exhibited inhibition. In contrast, non-GABAergic nonpyramidal neurons in the deep layer tended to be strongly inhibited by multiple different odorants. Our findings suggest that odor representation is accomplished by both broadly tuned and narrow-tuned PNs in the aPCX of awake animals. In addition, various types of interneurons may play different roles in the intracortical processing of olfactory information.


Comparison of uncultured marrow mononuclear cells and culture-expanded mesenchymal stem cells in 3D collagen-chitosan microbeads for orthopedic tissue engineering.
Stem cell-based therapies have shown promise in enhancing repair of bone and cartilage. Marrow-derived mesenchymal stem cells (MSC) are typically expanded in vitro to increase cell number, but this process is lengthy, costly, and there is a risk of contamination and altered cellular properties. Potential advantages of using fresh uncultured bone marrow mononuclear cells (BMMC) include heterotypic cell and paracrine interactions between MSC and other marrow-derived cells including hematopoietic, endothelial, and other progenitor cells. In the present study, we compared the osteogenic and chondrogenic potential of freshly isolated BMMC to that of cultured-expanded MSC, when encapsulated in three-dimensional (3D) collagen-chitosan microbeads. The effect of low and high oxygen tension on cell function and differentiation into orthopedic lineages was also examined. Freshly isolated rat BMMC (25 × 10(6) cells/mL, containing an estimated 5 × 10(4) MSC/mL) or purified and culture-expanded rat bone marrow-derived MSC (2 × 10(5) cells/mL) were added to a 65-35 wt% collagen-chitosan hydrogel mixture and fabricated into 3D microbeads by emulsification and thermal gelation. Microbeads were cultured in control MSC growth media in either 20% O2 (normoxia) or 5% O2 (hypoxia) for an initial 3 days, and then in control, osteogenic, or chondrogenic media for an additional 21 days. Microbead preparations were evaluated for viability, total DNA content, calcium deposition, and osteocalcin and sulfated glycosaminoglycan expression, and they were examined histologically. Hypoxia enhanced initial progenitor cell survival in fresh BMMC-microbeads, but it did not enhance osteogenic potential. Fresh uncultured BMMC-microbeads showed a similar degree of osteogenesis as culture-expanded MSC-microbeads, even though they initially contained only 1/10th the number of MSC. Chondrogenic differentiation was not strongly supported in any of the microbead formulations. This study demonstrates the microbead-based approach to culturing and delivering cells for tissue regeneration, and suggests that fresh BMMC may be an alternative to using culture-expanded MSC for bone tissue engineering.


Bimanual microincision cataract surgery with implantation of an Akreos MI6O lens--one year follow-up.
To evaluate visual outcome and complications after bimanual microincision cataract surgery (MICS) with implantation of an Akreos MI60 intraocular lens (IOL). This study comprised 40 eyes of 22 patients after MICS. The IOL was implanted through a 1.9 mm or smaller clear corneal incision. One year after operation, uncorrected and best corrected distance visual acuity, best corrected near visual acuity (logMAR), contrast sensitivities (CS) (CSV-1000), induce astigmatism, complications and patient satisfaction were analyzed. The Akreos lens was implanted through mean incision of 1.8 + 0.01 mm. One year after surgery the mean uncorrected and the best corrected distance visual acuity (UDVA, BCDVA) and the best corrected near visual acuity (BCNVA) significantly improved (UDVA: preoperatively--0.7 +/- 0.6; postoperatively--0.04 +/- 0.17; p < 0.005; BCDVA: preoperatively 0.61 +/- 0.6; postoperatively--0.01 +/- 0.12; p < 0.001; BCNVA: preoperatively--0.56 +/- 0.42; postoperatively--0.0 +/- 0; p < 0.005). CS in photopic adaptation were within normal age-matched limits. One year after surgery, there was not significant induced astigmatism measured by vector analysis. There were no serious postoperative complications. Visual significant PCO was observed in 1 patient. All the patients were highly satisfied with the quality of the vision. Our results show that MICS with Akreos MI60 lens implantation is a procedure which let receive very good visual function results, fast visual rehabilitation as well as high patients' satisfaction.


Photodynamic Diagnosis-guided Transurethral Resection of Bladder Tumour in Participants with a First Suspected Diagnosis of Intermediate- or High-risk Non-muscle-invasive Bladder Cancer: Cost-effectiveness Analysis Alongside a Randomised Controlled Trial.
Recurrence of non-muscle-invasive bladder cancer (NMIBC) is common after transurethral resection of bladder tumour (TURBT). Photodynamic diagnosis (PDD) may reduce recurrence. PDD uses a photosensitiser in the bladder that causes the tumour to fluoresce to guide resection. PDD provides better diagnostic accuracy and allows more complete tumour resection. To estimate the economic efficiency of PDD-guided TURBT (PDD-TURBT) in comparison to white light-guided TURNT (WL-TURBT) in individuals with a suspected first diagnosis of NMIBC at intermediate or high risk of recurrence on the basis of routine visual assessment before being scheduled for TURBT. This is a health economic evaluation alongside a pragmatic, open-label, parallel-group randomised trial from a societal perspective. A total of 493 participants (aged ≥16 yr) were randomly allocated to PDD-TURBT (n = 244) or WL-TURBT (n = 249) in 22 UK National Health Service hospitals. Cost effectiveness ratios were based on the use of health care resources associated with PDD-TURBT and WL-TURBT and quality-adjusted life years (QALYs) gained within the trial. Uncertainties in key parameters were assessed using sensitivity analyses. On the basis of the use of resources driven by the trial protocol, the incremental cost effectiveness of PDD-TURBT in comparison to WL-TURBT was not cost saving. At 3 yr, the total cost was £12 881 for PDD-TURBT and £12 005 for WL-TURBT. QALYs at three years were 2.087 for PDD-TURBT and 2.094 for WL-TURBT. The probability that PDD-TURBT is cost effective was never >30% above the range of societal cost-effectiveness thresholds. There was no evidence of a difference in either costs or QALYs over 3-yr follow-up between PDD-TURBT and WL-TURBT in individuals with suspected intermediate- or high-risk NMIBC. PDD-TURBT is not supported for the management of primary intermediate- or high-risk NMIBC. We assessed overall costs for two approaches for removal of bladder tumours in noninvasive cancer and measured quality-adjusted life years gained for each. We found that use of a photosensitiser in the bladder was not more cost effective than use of white light only during tumour removal.


A branching morphogenesis program governs embryonic growth of the thyroid gland.
The developmental program that regulates thyroid progenitor cell proliferation is largely unknown. Here, we show that branching-like morphogenesis is a driving force to attain final size of the embryonic thyroid gland in mice. Sox9, a key factor in branching organ development, distinguishes Nkx2-1+ cells in the thyroid bud from the progenitors that originally form the thyroid placode in anterior endoderm. As lobes develop the thyroid primordial tissue branches several generations. Sox9 and Fgfr2b are co-expressed distally in the branching epithelium prior to folliculogenesis. The thyroid in Fgf10 null mutants has a normal shape but is severely hypoplastic. Absence of Fgf10 leads to defective branching and disorganized angiofollicular units although Sox9/Fgfr2b expression and the ability of cells to differentiate and form nascent follicles are not impaired. These findings demonstrate a novel mechanism of thyroid development reminiscent of the Fgf10-Sox9 program that characterizes organogenesis in classical branching organs, and provide clues to aid understanding of how the endocrine thyroid gland once evolved from an exocrine ancestor present in the invertebrate endostyle.


Characteristic Factors Affecting Oral Pigmentation in Passive Smoker Children.
Smoking affects not only smokers themselves, but also the people around them. 700 million children are exposed to second hand tobacco worldwide. One of the adverse effects of being a passive smoker is oral pigmentation. This study was conducted to evaluate the association between smoking of a parent at home and oral pigmentation in children, and the characteristic factors affecting that. In this retrospective cohort study, 140 healthy children aged 4 to 10 (mean age= 6.68±1.60), 70 with smoker parent and 70 without smoker parents, were examined for oral pigmentation. Environmental factors were evaluated by asking the parents to fill a questionnaire. Data were analyzed using Chi-square test, Fisher's exact test, Logistic regression, and Spearman scale. There was a meaningful relationship between having a smoker parent and oral pigmentation (p= 0.0001). Spearman's correlation showed parents' duration of cigarette smoking and the number of cigarettes per day could meaningfully affect the severity of oral pigmentation (R=0.329). The study did not find a statistical relationship between oral pigmentation in passive smoking and gender or house area. Children exposed to secondhand tobacco are at more risk for oral pigmentation. Its severity depends on duration of cigarette smoking and the number of cigarettes per day.


Discussion based on analysis of the suicide rate and the average disposable income per household in Japan.
Suicide is one of the major social issues in Japan. According to a report of the National Policy Agency, there were approximately 22 000 to 24 000 annual suicides between 1994 and 1997 and there have been over 30 000 annual suicides in Japan since 1998. For these reasons, we think it is important to discuss the economic factor related to suicides in recent years. In this study, we examined suicide rates and the average disposable income per household in Japan in the last 15 years (ie 1994-2008) and discuss the statistical analysis of the average disposable income per household and the associated suicide rates. During the research period, annual suicide rates per 100 000 population in Japan ranged from 16.9 to 25.5 among the total population, from 23.1 to 38.0 among men, and from 10.9 to 14.7 among women. The annual average disposable income per household (ten thousand yen) ranged from 424.0 to 549.9. The average disposable income per household was related to the suicide rate among the total population and among men. The average disposable income per household was not related to the suicide rate among women. We believe that this discussion will be useful in developing specific suicide preventive measures.