Synthesis and characterization of gold glyconanoparticles functionalized with sugars of sweet Sorghum syrup.
Gold glyconanoparticles were synthesized by a simple, rapid, and eco-friendly method by using sweet Sorghum syrup for application in biomedicine and biotechnology. The nanostructures of the prepared gold nanoparticles were confirmed by using UV-visible absorbance, TEM, SAED, FTIR, EDAX, XRD, and photoluminescence analyses. The formation of gold nanoparticles at both room and boiling temperatures and kinetics of the reaction were monitored by UV-visible spectroscopy and TEM studies. TEM analysis revealed that the obtained nanoparticles were mono-dispersed and spherical in shape with an average particle size of 7 nm. The size of the nanoparticles was influenced by the concentration of Sorghum syrup. The presence of elemental gold was confirmed by EDAX analysis. Based on the FTIR analysis, it was observed that the sugars present in the Sorghum syrup possibly acts as capping agents. The zeta potential analysis revealed that the glyconanoparticles were negatively charged with a potential of -25 mV. The XRD and SAED patterns also suggest that the nanoparticles were crystalline in nature and these particles were found to exhibit visible photoluminescence. Fructose and glucose present in sweet Sorghum syrup were demonstrated as responsible sugars for the reduction of gold ions, and sucrose stabilized the formed nanoparticles. The proposed mechanism for the formation and stabilization of gold glyconanoparticles is based on the phenomenon of "macromolecular crowding." This is the first report on the use of sweet Sorghum syrup for the green synthesis of gold glyconanoparticles at both room and boiling temperatures.


Laryngeal dysplasia, demographics, and treatment: a single-institution, 20-year review.
Laryngeal dysplasia is a common disease entity that remains clinically frustrating because functional outcomes are balanced against oncologic results. Understanding evolution in dysplasia demographics, treatment, and progression rates may inform better therapy in the future. To review laryngeal dysplasia cases at a single institution during the last 20 years and identify changes in patient demographics, categorize treatment approaches, and review rates of progression to cancer. In this retrospective medical record review, patients with laryngeal dysplasia treated at an academic medical center were identified on review of pathology records. Patients were organized by date of dysplasia diagnosis, divided into 2 groups (group 1, January 1, 1993, through December 31, 2002; group 2, January 1, 2003, through December 31, 2012), and compared against one another. Age at diagnosis, sex, type of treatment, and progression to malignant disease were analyzed from one period to the next. A total of 107 patients were identified through review of pathology databases. Progression of dysplasia to cancer remained roughly stable across periods, at 8.8% and 8.0%, respectively. Mean age at diagnosis decreased from 68.7 to 61.7 years over time, with a statistically significant trend toward presentation at younger ages. The male to female ratio was 3.75 in group 1 and 3.17 in group 2, with a trend toward a greater proportion of females over time that did not reach statistical significance. Use of radiotherapy remained stable across groups, with increased use of microflap excision techniques and laser treatment (especially photoangiolytic lasers) in group 2. Overall, progression of laryngeal dysplasia to cancer has remained stable during the past 20 years at a rate of approximately 8%. Although laryngeal dysplasia remains a disease predominantly found in males, there is a demographic trend toward diagnosis at earlier ages. Treatment choices may slowly be changing over time, although multi-institutional studies may be required to better categorize this shift.


Ginger extract activates caspase independent paraptosis in cancer cells via ER stress, mitochondrial dysfunction, AIF translocation and DNA damage.
The rhizome of ginger (Zingiber officinale) a common culinary agent is also known for its medicinal activity. We have earlier reported that pure 6-shogaol, an important component of ginger induces paraptosis in triple negative breast cancer (MDA-MB-231) and non small cell lung (A549) cancer cells. However, the chemopreventive potential of the whole ginger extract in food remains to be elucidated. Here, we demonstrate for the first time that ginger extract (GE) triggers similar anticancer activity/paraptosis against the same cell lines but through different molecular mechanisms. Q-TOF LC-MS analysis of the extract showed the presence of several other metabolites along with 6-shogaol and 6-gingerol. GE induces cytoplasmic vacuolation through ER stress and dilation of the ER. Drastic decrease in the mitochondrial membrane potential and ATP production along with the excess generation of ROS contributed to mitochondrial dysfunction. Consequently, GE caused the translocation of apoptosis inducing factor to the nucleus leading to the fragmentation of DNA. Taken together, these show a novel mechanism for ginger extract induced cancer cell death that can be of potential interest for cancer preventive strategies.


Technique considerations and complication management in transurethral resection of the prostate and photoselective vaporization of the prostate.
The prevalence of lower urinary tract symptoms (LUTS) and benign prostatic hyperplasia (BPH) increases with age. While a variety of treatments are available for these men, endoscopic treatments are generally preferred for men with small to moderate size glands. Novel treatment options are continually introduced into this large market. However, the practicing urologist should have a well tested surgical option in regular practice that is applicable to a wide range of patients. Herein we discuss two well recognized surgical options that can be used for the majority of men with LUTS due to BPH who have failed medical management.


Palmitoylethanolamide Reduces Neuropsychiatric Behaviors by Restoring Cortical Electrophysiological Activity in a Mouse Model of Mild Traumatic Brain Injury.
Traumatic brain injury (TBI) represents a major public health problem, which is associated with neurological dysfunction. In severe or moderate cases of TBI, in addition to its high mortality rate, subjects may encounter diverse behavioral dysfunctions. Previous reports suggest that an association between TBI and chronic pain syndromes tends to be more common in patients with mild forms of brain injury. Despite causing minimal brain damage, mild TBI (mTBI) often leads to persistent psychologically debilitating symptoms, which can include anxiety, various forms of memory and learning deficits, and depression. At present, no effective treatment options are available for these symptoms, and little is known about the complex cellular activity affecting neuronal activity that occurs in response to TBI during its late phase. Here, we used a mouse model to investigate the effect of Palmitoylethanolamide (PEA) on both the sensorial and neuropsychiatric dysfunctions associated with mTBI through behavioral, electrophysiological, and biomolecular approaches. Fourteen-day mTBI mice developed anxious, aggressive, and reckless behavior, whilst depressive-like behavior and impaired social interactions were observed from the 60th day onward. Altered behavior was associated with changes in interleukin 1 beta (IL-1β) expression levels and neuronal firing activity in the medial prefrontal cortex. Compared with vehicle, PEA restored the behavioral phenotype and partially normalized the biochemical and functional changes occurring at the supraspinal level. In conclusion, our findings reveal some of the supraspinal modifications responsible for the behavioral alterations associated with mTBI and suggest PEA as a pharmacological tool to ameliorate neurological dysfunction induced by the trauma.


Inhibition of in vitro NZB antibody responses by cyclosporine.
To characterize functional abnormalities of B cells in murine autoimmunity, the effect of cyclosporine (CsA) on antibody responses of NZB and control BALB/c spleen cells was investigated in vitro. Under conditions of high cell density, both NZB and BALB/c spleen cells spontaneously produced IgM and IgM anti-DNA, although NZB responses were greater in magnitude and demonstrated at lower cell densities. In cultures at 5 x 10(6) cells/ml, CsA preferentially inhibited anti-DNA production by cells of both strains. However, to achieve similar levels of inhibition of both responses, significantly higher concentrations of CsA were required for cultures of NZB compared to BALB/c cells. These results support previous experiments indicating enhanced sensitivity to CsA as a pharmacological marker of autoantibody producing B cells. They further suggest that B cell activation in certain forms of autoimmunity may be associated with decreased sensitivity to immunosuppressive agents.


Effect of increasing norepinephrine dosage on regional blood flow in a porcine model of endotoxin shock.
To evaluate the effect of a norepinephrine-induced differential increase in mean arterial pressure on splanchnic and renal perfusion in a porcine model of volume-resuscitated endotoxic shock. Prospective, controlled, acute interventional study. Animal research laboratory. Fourteen landrace pigs, seven treated with norepinephrine and seven used as endotoxemic controls. In an acute endotoxic shock model, norepinephrine was used to reverse hypotension in seven fluid-resuscitated pigs, anesthetized with alpha-chloralose and equipped with flow probes around the portal vein and renal artery, renal and jejunal mucosal laser Doppler flowmetry, and jejunal tonometry. Mean arterial pressure was increased by 10 and then 20 mm Hg above the shock level with norepinephrine. Seven shocked, fluid-resuscitated only animals served as the comparison group. Measurements were performed before 2-hr endotoxin infusion and at the end of each increased level of mean arterial pressure. Raising mean arterial pressure with norepinephrine by 10 mm Hg significantly increased cardiac output, systemic oxygen extraction, and portal vein blood flow; stabilized metabolic acidosis; and tended to restore renal and jejunal mucosal flows to preshock levels. Increasing mean arterial pressure by 20 mm Hg further increased cardiac output and oxygen delivery but without improving portal vein, renal artery, and jejunal mucosal blood flows. Norepinephrine, administered to increase mean arterial pressure by 10 mm Hg in an acute model of volume-resuscitated endotoxic shock, improved systemic and regional perfusion. The administration of norepinephrine to increase mean arterial pressure 20 mm Hg above shock did not increase renal and splanchnic blood flows, despite an enhanced cardiac output.


Linked linear amplification: a new method for the amplification of DNA.
Linked Linear Amplification (LLA) is a new nucleic acid amplification method that uses multiple cycles of primer extension reactions. The presence of nonreplicable elements in LLA primers renders primer extension products unusable as templates for further amplification, leading to linear accumulation of products. Through the use of nested primers, linear reactions can be "linked", providing total amplification yields comparable to those obtained by PCR. The LLA model predicts (a) that amplification yield will approach that of PCR as the number of primers increases and (b) that the unique composition of LLA products will give lower carryover amplification efficiency compared with PCR. To test these hypotheses, the human ss-globin gene was amplified by 10-, 14-, or 18-primer LLA and the yield was compared with PCR. Carryover contamination was simulated by reamplifying a dilution series of LLA or PCR products. To demonstrate the clinical utility of the method, LLA coupled with allele-specific oligonucleotide (ASO) capture was used to detect the factor V Leiden mutation in a panel of 111 DNA samples. Fourteen- and 18-primer LLA gave amplification yields comparable to PCR. However, LLA carryover amplification efficiency was four orders of magnitude lower than that of PCR. The LLA-ASO assay detected the correct factor V Leiden genotype in all 111 samples. LLA is a robust target amplification method that is comparable to PCR in yield. However, LLA is more resistant to false results caused by carryover amplicon contamination.


Response to rivastigmine transdermal patch or memantine plus rivastigmine patch is affected by apolipoprotein E genotype in Alzheimer patients.
The apolipoprotein E (APOE) genotype in response to pharmacological treatments in patients with Alzheimer's disease (AD) remains a matter of controversy. This analysis investigated the effect of the APOE genotype on the clinical response to rivastigmine transdermal patch monotherapy or memantine plus rivastigmine patch in patients with mild to moderate AD. Two hundred and six (n = 206) patients with probable AD and Mini-Mental State Examination (MMSE) scores of 10-20 were randomized to rivastigmine patch monotherapy or memantine plus rivastigmine patch for 24 weeks. Of the 206 patients with probable AD, 146 patients who consented to genetic testing for APOE were included and assessed for this subgroup study. There were no significant differences on MMSE, NPI, ADAS-cog, ADCS-ADL, CDR-SB, NPI and FAB between rivastigmine patch monotherapy and memantine plus rivastigmine patch according to the APOE genotype. However, patients with moderately severe AD (MMSE ≤15) who were APOE ε4 carriers showed higher responder rates on ADCS-ADL with memantine plus rivastigmine patch compared to rivastigmine patch monotherapy. Moderately severe AD patients with the APOE ε4 allele may respond more favorably to memantine plus rivastigmine patch than ε4 noncarriers.


Spatial inequalities in skilled attendance at birth in Ghana: a multilevel analysis integrating health facility databases with household survey data.
This study aimed at using survey data to predict skilled attendance at birth (SBA) across Ghana from healthcare quality and health facility accessibility. Through a cross-sectional, observational study, we used a random intercept mixed effects multilevel logistic modelling approach to estimate the odds of having SBA and then applied model estimates to spatial layers to assess the probability of SBA at high-spatial resolution across Ghana. We combined data from the Demographic and Health Survey (DHS), routine birth registers, a service provision assessment of emergency obstetric care services, gridded population estimates and modelled travel time to health facilities. Within an hour's travel, 97.1% of women sampled in the DHS could access any health facility, 96.6% could reach a facility providing birthing services, and 86.2% could reach a secondary hospital. After controlling for characteristics of individual women, living in an urban area and close proximity to a health facility with high-quality services were significant positive determinants of SBA uptake. The estimated variance suggests significant effects of cluster and region on SBA as 7.1% of the residual variation in the propensity to use SBA is attributed to unobserved regional characteristics and 16.5% between clusters within regions. Given the expansion of primary care facilities in Ghana, this study suggests that higher quality healthcare services, as opposed to closer proximity of facilities to women, is needed to widen SBA uptake and improve maternal health.


Rapid purification and characterization of γ-glutamyl-transpeptidase from shiitake mushroom (Lentinus edodes).
γ-Glutamyl-transpeptidase (GGT) is one of the important enzymes in the pathway of odor formation in shiitake mushroom (Lentinus edodes). Rapid purification and characterization of GGT from shiitake mushroom were studied in this work. The GGT was purified 179-fold after 3 primary steps: precipitation by ammonium sulfate, isolation by Phenyl Sepharose 6 FF, and desalting by Sephadex G-25. The enzyme, consisting of a small and a large subunit with Mr 28 KDa and 60 KDa, respectively, is composed of 17 kinds of amino acids with the ratio of basic and acidic residues 1: 1.84, and its secondary structure was also determined by Fourier transform infrared spectroscopy. The properties of GGT were studied with γ-glutamyl-p-nitroanilide as the substrate. The results showed Km value of 2.601μM, optimal temperature of 40 °C, and isoelectric point of 6.4. In addition, the activity of GGT was promoted by Na⁺, K⁺, and Ca²⁺ and inhibited by Cu²⁺, Ag⁺, Zn²⁺, and Fe³⁺. In this work, the γ-glutamyl-transpeptidase (GGT) from shiitake mushroom was purified with a simple scheme. Through the characterization of GGT, the relationship between endogenous formaldehyde and odor formation is to be clarified and assist in finding means of formaldehyde control in shiitake mushroom.


Drosophila hemopoiesis and cellular immunity.
In Drosophila melanogaster larvae, three classes of circulating cellular immune surveillance cells (hemocytes) can be identified: plasmatocytes, crystal cells, and lamellocytes. Plasmatocytes are professional phagocytes most similar to the mammalian monocyte/macrophage lineage and make up approximately 95% of circulating hemocytes. The other approximately 5% of circulating hemocytes consists of crystal cells, which secrete components necessary for the melanization of invading organisms, as well as for wound repair. A third cell type known as lamellocytes are rarely seen in healthy larvae and are involved in the encapsulation of invading pathogens. There are no obvious mammalian counterparts for crystal cells or lamellocytes, and there is no equivalent to the lymphoid lineage in insects. In this review, I will discuss what is currently known about Drosophila hemopoiesis and the cellular immune response and where possible compare it to vertebrate mechanisms.


Development and validation of the Family Decision-Making Self-Efficacy Scale.
Several studies have reported high levels of distress in family members who have made health care decisions for loved ones at the end of life. A method is needed to assess the readiness of family members to take on this important role. Therefore, the purpose of this study was to develop and validate a scale to measure family member confidence in making decisions with (conscious patient scenario) and for (unconscious patient scenario) a terminally ill loved one. On the basis of a survey of family members of patients with amyotrophic lateral sclerosis (ALS) enriched by in-depth interviews guided by Self-Efficacy Theory, we developed six themes within family decision making self-efficacy. We then created items reflecting these themes that were refined by a panel of end-of-life research experts. With 30 family members of patients in an outpatient ALS and a pancreatic cancer clinic, we tested the tool for internal consistency using Cronbach's alpha and for consistency from one administration to another using the test-retest reliability assessment in a subset of 10 family members. Items with item to total scale score correlations of less than .40 were eliminated. A 26-item scale with two 13-item scenarios resulted, measuring family self-efficacy in decision making for a conscious or unconscious patient with a Cronbach's alphas of .91 and .95, respectively. Test-retest reliability was r = .96, p = .002 in the conscious senario and r = .92, p = .009 in the unconscious scenario. The Family Decision-Making Self-Efficacy Scale is valid, reliable, and easily completed in the clinic setting. It may be used in research and clinical care to assess the confidence of family members in their ability to make decisions with or for a terminally ill loved one.


Identification of a sudden cardiac death susceptibility locus at 2q24.2 through genome-wide association in European ancestry individuals.
Sudden cardiac death (SCD) continues to be one of the leading causes of mortality worldwide, with an annual incidence estimated at 250,000-300,000 in the United States and with the vast majority occurring in the setting of coronary disease. We performed a genome-wide association meta-analysis in 1,283 SCD cases and >20,000 control individuals of European ancestry from 5 studies, with follow-up genotyping in up to 3,119 SCD cases and 11,146 controls from 11 European ancestry studies, and identify the BAZ2B locus as associated with SCD (P = 1.8×10(-10)). The risk allele, while ancestral, has a frequency of ~1.4%, suggesting strong negative selection and increases risk for SCD by 1.92-fold per allele (95% CI 1.57-2.34). We also tested the role of 49 SNPs previously implicated in modulating electrocardiographic traits (QRS, QT, and RR intervals). Consistent with epidemiological studies showing increased risk of SCD with prolonged QRS/QT intervals, the interval-prolonging alleles are in aggregate associated with increased risk for SCD (P = 0.006).


Gram-typing of mastitis bacteria in milk samples using flow cytometry.
Fast identification of pathogenic bacteria in milk samples from cows with clinical mastitis is central to proper treatment. In Denmark, time to bacterial diagnosis is typically 24 to 48 h when using traditional culturing methods. The PCR technique provides a faster and highly sensitive identification of bacterial pathogens, although shipment of samples to diagnostic laboratories delays treatment decisions. Due to the lack of fast on-site tests that can identify the causative pathogens, antibiotic treatments are often initiated before bacterial identification. The present study describes a flow cytometry-based method, which can detect and distinguish gram-negative and gram-positive bacteria in mastitis milk samples. The differentiation was based on bacterial fluorescence intensities upon labeling with biotin-conjugated wheat germ agglutinin and acridine orange. Initially 19 in-house bacterial cultures (4 gram-negative and 15 gram-positive strains) were analyzed, and biotin-conjugated wheat germ agglutinin and acridine orange florescence intensities were determined for gram-negative and gram-positive bacteria, respectively. Fluorescence cut-off values were established based on receiver operating characteristic curves for the 19 bacterial cultures. The method was then tested on 53 selected mastitis cases obtained from the department biobank (milk samples from 6 gram-negative and 47 gram-positive mastitis cases). Gram-negative bacteria in milk samples were detected with a sensitivity of 1 and a specificity of 0.74, when classification was based on the previously established cut-off values. However, when receiver operating characteristic curves were constructed for the 53 mastitis cases, results indicate that a sensitivity and specificity of 1 could be reached if cut-off values were reduced. This flow cytometry-based technique could potentially provide dairy farmers and attending veterinarians with on-site information on bacterial gram-type and prevent ineffective antimicrobial treatment in mastitis cases caused by gram-negative bacteria.


The cellular and molecular mechanisms of vertebrate lens development.
The ocular lens is a model system for understanding important aspects of embryonic development, such as cell specification and the spatiotemporally controlled formation of a three-dimensional structure. The lens, which is characterized by transparency, refraction and elasticity, is composed of a bulk mass of fiber cells attached to a sheet of lens epithelium. Although lens induction has been studied for over 100 years, recent findings have revealed a myriad of extracellular signaling pathways and gene regulatory networks, integrated and executed by the transcription factor Pax6, that are required for lens formation in vertebrates. This Review summarizes recent progress in the field, emphasizing the interplay between the diverse regulatory mechanisms employed to form lens progenitor and precursor cells and highlighting novel opportunities to fill gaps in our understanding of lens tissue morphogenesis.


RNA-Seq of Kaposi's sarcoma reveals alterations in glucose and lipid metabolism.
Kaposi's sarcoma-associated herpesvirus (KSHV) is the etiologic agent of Kaposi's sarcoma (KS). It is endemic in a number of sub-Saharan African countries with infection rate of >50%. The high prevalence of HIV-1 coupled with late presentation of advanced cancer staging make KS the leading cancer in the region with poor prognosis and high mortality. Disease markers and cellular functions associated with KS tumorigenesis remain ill-defined. Several studies have attempted to investigate changes of the gene profile with in vitro infection of monoculture models, which are not likely to reflect the cellular complexity of the in vivo lesion environment. Our approach is to characterize and compare the gene expression profile in KS lesions versus non-cancer tissues from the same individual. Such comparisons could identify pathways critical for KS formation and maintenance. This is the first study that utilized high throughput RNA-seq to characterize the viral and cellular transcriptome in tumor and non-cancer biopsies of African epidemic KS patients. These patients were treated anti-retroviral therapy with undetectable HIV-1 plasma viral load. We found remarkable variability in the viral transcriptome among these patients, with viral latency and immune modulation genes most abundantly expressed. The presence of KSHV also significantly affected the cellular transcriptome profile. Specifically, genes involved in lipid and glucose metabolism disorder pathways were substantially affected. Moreover, infiltration of immune cells into the tumor did not prevent KS formation, suggesting some functional deficits of these cells. Lastly, we found only minimal overlaps between our in vivo cellular transcriptome dataset with those from in vitro studies, reflecting the limitation of in vitro models in representing tumor lesions. These findings could lead to the identification of diagnostic and therapeutic markers for KS, and will provide bases for further mechanistic studies on the functions of both viral and cellular genes that are involved.


Human Immunization With a Polymorphic Malaria Vaccine Candidate Induced Antibodies to Conserved Epitopes That Promote Functional Antibodies to Multiple Parasite Strains.
Overcoming antigenic diversity is a key challenge in the development of effective Plasmodium falciparum malaria vaccines. Strategies that promote the generation of antibodies targeting conserved epitopes of vaccine antigens may provide protection against diverse parasites strains. Understanding differences between vaccine-induced and naturally acquired immunity is important to achieving this goal. We analyzed antibodies generated in a phase 1 human vaccine trial, MSP2-C1, which included 2 allelic forms of MSP2, an abundant vaccine antigen on the merozoite surface. Vaccine-induced responses were assessed for functional activity against multiple parasite strains, and cross-reactivity of antibodies was determined using competition ELISA and epitope mapping approaches. Vaccination induced cytophilic antibody responses with strain-transcending opsonic phagocytosis and complement-fixing function. In contrast to antibodies acquired via natural infection, vaccine-induced antibodies were directed towards conserved epitopes at the C-terminus of MSP2, whereas naturally acquired antibodies mainly targeted polymorphic epitopes. Functional activity of C-terminal-targeted antibodies was confirmed using monoclonal antibodies that promoted opsonic phagocytosis against multiple parasite strains. Vaccination generated markedly different responses to polymorphic antigens than naturally acquired immunity and targeted conserved functional epitopes. Induction of antibodies targeting conserved regions of malaria antigens provides a promising vaccine strategy to overcome antigenic diversity for developing effective malaria vaccines.


Next-generation COVID-19 vaccines: Opportunities for vaccine development and challenges in tackling COVID-19.
The ongoing COVID-19 pandemic, caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), has become a global threat. Although non-pharmaceutical interventions have been rigorously and widely implemented, living conditions caused by the pandemic will last until highly effective vaccines are successfully improved and globally administered. Several first-generation COVID-19 vaccines were approved at the end of 2020. However, the COVID-19 pandemic is persisting worldwide. To be clear, the efficiency and the coverage of current vaccines are insufficient, but newly emerging and rapidly spreading variants are the most pressing concern. A second-generation COVID-19 vaccine worth mentioning, NVX-CoV2373, has demonstrated 90% overall efficacy as well as a high level of efficacy against circulating variants in Phase 3 clinical trials. Currently, NVX-CoV2373 is the only vaccine that has proven successful against variants during Phase 3/4 trials. Therefore, developing the next generation of vaccines is a promising strategy to ultimately prevail against SARS-CoV-2. This review provides up-to-date information on COVID-19 vaccines in terms of their efficacy and new platforms and the progression of COVID-19 vaccination. Moreover, this review also summarizes the efficacy of approved COVID-19 vaccines against variants. Lastly, this review highlights the global challenges for COVID-19 vaccines in development and vaccination, and it discusses opportunities for development of future COVID-19 vaccines and vaccination coverage.


Regional and Microenvironmental Scale Characterization of the Zostera muelleri Seagrass Microbiome.
Seagrasses are globally distributed marine plants that represent an extremely valuable component of coastal ecosystems. Like terrestrial plants, seagrass productivity and health are likely to be strongly governed by the structure and function of the seagrass microbiome, which will be distributed across a number of discrete microenvironments within the plant, including the phyllosphere, the endosphere and the rhizosphere, all different in physical and chemical conditions. Here we examined patterns in the composition of the microbiome of the seagrass Zostera muelleri, within six plant-associated microenvironments sampled across four different coastal locations in New South Wales, Australia. Amplicon sequencing approaches were used to characterize the diversity and composition of bacterial, microalgal, and fungal microbiomes and ultimately identify "core microbiome" members that were conserved across sampling microenvironments. Discrete populations of bacteria, microalgae and fungi were observed within specific seagrass microenvironments, including the leaves and roots and rhizomes, with "core" taxa found to persist within these microenvironments across geographically disparate sampling sites. Bacterial, microalgal and fungal community profiles were most strongly governed by intrinsic features of the different seagrass microenvironments, whereby microscale differences in community composition were greater than the differences observed between sampling regions. However, our results showed differing strengths of microbial preferences at the plant scale, since this microenvironmental variability was more pronounced for bacteria than it was for microalgae and fungi, suggesting more specific interactions between the bacterial consortia and the seagrass host, and potentially implying a highly specialized coupling between seagrass and bacterial metabolism and ecology. Due to their persistence within a given seagrass microenvironment, across geographically discrete sampling locations, we propose that the identified "core" microbiome members likely play key roles in seagrass physiology as well as the ecology and biogeochemistry of seagrass habitats.


Elliptically polarized light for depth resolved optical imaging.
It is shown that using elliptically polarized light permits selecting well-defined subsurface volumes in a turbid medium. This suggests the possibility of probing biological tissues at specific depths. First, we present the method and preliminary results obtained on an Intralipid phantom. We next report on the method's performance on a biological phantom (chicken breast) and, finally, on the exposed cortex of an anesthetized rat.


Chemerin-9, a potent agonist of chemerin receptor (ChemR23), prevents atherogenesis.
Plasma levels of chemerin, an adipocytokine produced from the adipose tissues and liver, are associated with metabolic syndrome and coronary artery disease (CAD). Chemerin and its analog, chemerin-9, are known to bind to their receptor, ChemR23. However, whether chemerin and chemerin-9 affect atherogenesis remains to be elucidated. We investigated the expression of chemerin and ChemR23 in human coronary arteries and cultured human vascular cells. The effects of chemerin and chemerin-9 on atheroprone phenomena were assessed in human THP1 monocytes, human umbilical vein endothelial cells (HUVECs), and human aortic smooth muscle cells (HASMCs) and aortic lesions in Apoe-/- mice. In patients with CAD, a small amount of ChemR23, but not chemerin, was expressed within atheromatous plaques in coronary arteries. Chemerin and ChemR23 were expressed at high levels in THP1 monocytes, THP1-derived macrophages, and HUVECs; however, their expression in HASMCs was weak. Chemerin and chemerin-9 significantly suppressed the tumor necrosis factor-α (TNF-α)-induced mRNA expression of adhesion and pro-inflammatory molecules in HUVECs. Chemerin and chemerin-9 significantly attenuated the TNF-α-induced adhesion of THP1 monocytes to HUVECs and macrophage inflammatory phenotype. Chemerin and chemerin-9 suppressed oxidized low-density lipoprotein (oxLDL)-induced macrophage foam cell formation associated with down-regulation of CD36 and up-regulation of ATP-binding cassette transporter A1 (ABCA1). In HASMCs, chemerin and chemerin-9 significantly suppressed migration and proliferation without inducing apoptosis. In the Apoe-/- mice, a 4-week infusion of chemerin-9 significantly decreased the areas of aortic atherosclerotic lesions by reducing intraplaque macrophage and SMC contents. Our results indicate that chemerin-9 prevents atherosclerosis. Therefore, the development of chemerin analogs/ChemR23 agonists may serve as a novel therapeutic target for atherosclerotic diseases.


Enhanced light output power of quantum cascade lasers from a tilted front facet.
We present a technique for enhancing the light output power of quantum cascade lasers (QCLs) by tilting of the front facet, which leads to a change of the modal reflectivity, resulting in an asymmetric light intensity distribution along the laser cavity. This asymmetry provides most of the light being emitted through one facet of the laser. An experimental study of threshold current, slope efficiency and light output power as a function of the front facet angles were performed and compared to conventional QCLs. The lasers with a front facet angle of 8° shows a 20% improved power output from the front facet.


Baicalein protects C6 glial cells against hydrogen peroxide-induced oxidative stress and apoptosis through regulation of the Nrf2 signaling pathway.
Baicalein, a flavonoid originally obtained from the roots of Scutellaria baicalensis Georgi, has been reported to possess various biological properties. Although several studies have demonstrated the anti-oxidative activity of baicalein, its neuroprotective mechanisms have not been clearly established. The present study aimed to detect the effects of baicalein against hydrogen peroxide (H2O2)-induced neuronal damage in C6 glial cells and to investigate the molecular mechanisms involved in this process. The results demonstrated that baicalein effectively inhibited H2O2-induced growth and reactive oxygen species (ROS) generation. We noted that Baicalein also attenuated the H2O2‑induced formation of comet tail, phosphorylation of p-γH2A.X, loss of mitochondrial membrane potential (MMP or ΔΨm), and changes to apoptosis‑related protein expression, which suggests that it can prevent H2O2‑induced cellular DNA damage and apoptotic cell death. Furthermore, treatment with baicalein effectively induced the expression of nuclear factor-erythroid 2-related factor 2 (Nrf2) as well as heme oxygenase-1 (HO-1) and thioredoxin reductase 1 (TrxR1) in a concentration and time-dependent manner. Moreover, the protective effects of baicalein against H2O2‑induced DNA damage and apoptosis were abolished by zinc protoporphyrin (ZnPP) IX, a HO-1 inhibitor, and auranofin, a TrxR inhibitor. In addition, we noted that the cytoprotective effects of baicalein were attenuated by transient transfection with Nrf2-specific small interfering RNA (siRNA). The findings of our present study suggest that baicalein enhances cellular antioxidant defense capacity through the inhibition of ROS generation and the activation of the Nrf2 signaling pathway, thus protecting C6 cells from H2O2-induced neuronal damage.


Characterization of DNA damage inflicted by free radicals from a mutagenic sunscreen ingredient and its location using an in vitro genetic reversion assay.
We describe an in vitro approach to assessing the potential genotoxicity of illuminated sunscreens. The photomutagenic sunscreen Padimate-O attacks DNA on illumination with simulated sunlight, producing strand breaks and lesions that are labile to N,N'-dimethylethylenediamine but few, if any, cyclobutane dimers or other direct photoproducts. The damage can be completely suppressed by the free radical quenchers Tris, ethanol, mannitol and dimethylsulfoxide, which is commonly used as a solvent in conventional photomutagenicity assays. Using a genetic reversion assay that depends on regenerating beta-galactosidase activity in photodamaged plasmids we find that GC base pairs are particularly susceptible to attack by Padimate-O.


[The effects of hyperthermia on cerebral blood flow, metabolism and electroencephalogram].
In practice of systemic or local hyperthermia, it becomes very important to determine the tolerance of the brain to heat, along with the evaluation of the influence of heat on brain function. In this little known area, the study was made by examining the changes of blood pressure, cerebral blood flow (CBF), partial pressure of oxygen and pH in the cerebral cortex, electroencephalogram (EEG) and somatosensory evoked potential (SEP), under hyperthermia by experiment. Unanesthetized and immobilized adult rabbits were used in the experiments under controlled respiration. The head alone or the whole body was heated by bathing in hot air. In the head heating, the brain temperature could be heated up to 45 degrees C without extreme fall of the blood pressure, and meantime the aortic blood maintained under 43.5 degrees C at that condition. In the head heating case, the CBF increased to two times at 43 degrees C, two and a half times at 44 degrees C, three and a half times at 45 degrees C of the cortical temperature. As the cortical temperature raised up, the partial pressure of oxygen and pH of the cerebral cortex increased. The peak frequency of power spectrum of EEG increased as the temperature was raised. High amplitude rhythmic slow wave burst appeared at over 44 degrees C for 30 minutes, which soon changed to a flat pattern. In the whole body bathing, on the contrary, the animals died of abrupt fall of the arterial blood pressure when the temperature of the aortic blood was raised over 43.5 degrees C, and it was impossible to get the brain heated over 43 degrees C.(ABSTRACT TRUNCATED AT 250 WORDS)


How willing are patients to question healthcare staff on issues related to the quality and safety of their healthcare? An exploratory study.
One in 10 patients admitted to hospital will suffer an adverse event as a result of their medical treatment. A reduction in adverse events could happen if patients could be engaged successfully in monitoring their care. This study explored: (1) surgical patients' willingness to question healthcare staff about their treatment; (2) differences between patients' willingness to ask factual vs. challenging questions related to the quality and safety of their healthcare; (3) patient demographic characteristics that could affect patients' willingness to ask questions; and (4) the impact of doctors' instructions on patients' willingness to ask questions. Cross-sectional study using the Patient Willingness to Ask Safety Questions Survey (PWASQS). The PWASQS questions were devised in accordance with current patient safety initiatives aimed at encouraging patients to ask healthcare staff specific safety-related questions about their healthcare. The PWASQS includes factual questions (eg, "when can I return to my normal activities?") and challenging questions (eg, "have you washed your hands?"), and examines the impact of doctors' instructions on patients' willingness to ask challenging questions (eg, if instructed to by a doctor would you be willing to ask: "have you washed your hands?"). Data were analysed using non-parametric tests. An inner-city London teaching hospital. A convenience sample of 80 patients who had undergone surgery. Surgical patients were significantly more willing to ask: doctors factual versus challenging questions (z = 7.59, p<0.001); nurses factual versus challenging questions (z = 5.39, p<0.001); doctors versus nurses factual questions (z = 4.98, p<0.001); and, nurses versus doctors challenging questions (z = 4.40, p<0.001). Doctor's instructions to the patient increased patient willingness to challenge doctors (z = 6.56, p<0.001) and nurses (z = 6.15, p<0.001).Women, educated patients, and patients in employment, were more willing to ask questions (p<0.05). Surgical patients, particularly those who are men, less educated or unemployed are less willing to challenge healthcare staff regarding their care than to ask healthcare staff factual questions. Patient involvement strategies which take into account patient characteristics need to be developed for patients and staff in order to encourage patient involvement in this much neglected area.


Toward an online cognitive and emotional battery to predict treatment remission in depression.
To evaluate the performance of a cognitive and emotional test battery in a representative sample of depressed outpatients to inform likelihood of remission over 8 weeks of treatment with each of three common antidepressant medications. Outpatients 18-65 years old with nonpsychotic major depressive disorder (17 sites) were randomized to escitalopram, sertraline or venlafaxine-XR (extended release). Participants scored ≥12 on the baseline 16-item Quick Inventory of Depressive Symptomatology - Self-Report and completed 8 weeks of treatment. The baseline test battery measured cognitive and emotional status. Exploratory multivariate logistic regression models predicting remission (16-item Quick Inventory of Depressive Symptomatology - Self-Report score ≤5 at 8 weeks) were developed independently for each medication in subgroups stratified by age, sex, or cognitive and emotional test performance. The model with the highest cross-validated accuracy determined the participant proportion in each arm for whom remission could be predicted with an accuracy ≥10% above chance. The proportion for whom a prediction could be made with very high certainty (positive predictive value and negative predictive value exceeding 80%) was calculated by incrementally increasing test battery thresholds to predict remission/non-remission. The test battery, individually developed for each medication, improved identification of remitting and non-remitting participants by ≥10% beyond chance for 243 of 467 participants. The overall remission rates were escitalopram: 40.8%, sertraline: 30.3%, and venlafaxine-XR: 31.1%. Within this subset for whom prediction exceeded chance, test battery thresholds established a negative predictive value of ≥80%, which identified 40.9% of participants not remitting on escitalopram, 77.1% of participants not remitting on sertraline, and 38.7% of participants not remitting on venlafaxine-XR (all including 20% false negatives). The test battery identified about 50% of each medication group as being ≥10% more or less likely to remit than by chance, and identified about 38% of individuals who did not remit with ≥80% certainty. Clinicians might choose to avoid this specific medication in these particular patients.


Functional genomic analysis of midgut epithelial responses in Anopheles during Plasmodium invasion.
The malaria parasite Plasmodium must complete a complex developmental life cycle within Anopheles mosquitoes before it can be transmitted into the human host. One day after mosquito infection, motile ookinetes traverse the midgut epithelium and, after exiting to its basal site facing the hemolymph, develop into oocysts. Previously, we have identified hemolymph factors that can antagonize or promote parasite development. We profiled on a genomic scale the transcriptional responses of the A. gambiae midgut to P. berghei and showed that more than 7% of the assessed mosquito transcriptome is differentially regulated during invasion. The profiles suggested that actin- and microtubule-cytoskeleton remodeling is a major response of the epithelium to ookinete penetration. Other responses encompass components of innate immunity, extracellular-matrix remodeling, and apoptosis. RNAi-dependent gene silencing identified both parasite antagonists and agonists among regulators of actin dynamics and revealed that actin polymerization is inhibitory to the invading parasite. Combined transcriptional and reverse-genetic analysis further identified an unexpected dual role of the lipid-trafficking machinery of the hemolymph for both parasite and mosquito-egg development. We conclude that the determinants of malaria-parasite development in Anopheles include components not only of systemic humoral immunity but also of intracellular, local epithelial reactions. These results provide novel mechanistic insights for understanding malaria transmission in the mosquito vector.


AT_CHLORO, a comprehensive chloroplast proteome database with subplastidial localization and curated information on envelope proteins.
Recent advances in the proteomics field have allowed a series of high throughput experiments to be conducted on chloroplast samples, and the data are available in several public databases. However, the accurate localization of many chloroplast proteins often remains hypothetical. This is especially true for envelope proteins. We went a step further into the knowledge of the chloroplast proteome by focusing, in the same set of experiments, on the localization of proteins in the stroma, the thylakoids, and envelope membranes. LC-MS/MS-based analyses first allowed building the AT_CHLORO database (http://www.grenoble.prabi.fr/protehome/grenoble-plant-proteomics/), a comprehensive repertoire of the 1323 proteins, identified by 10,654 unique peptide sequences, present in highly purified chloroplasts and their subfractions prepared from Arabidopsis thaliana leaves. This database also provides extensive proteomics information (peptide sequences and molecular weight, chromatographic retention times, MS/MS spectra, and spectral count) for a unique chloroplast protein accurate mass and time tag database gathering identified peptides with their respective and precise analytical coordinates, molecular weight, and retention time. We assessed the partitioning of each protein in the three chloroplast compartments by using a semiquantitative proteomics approach (spectral count). These data together with an in-depth investigation of the literature were compiled to provide accurate subplastidial localization of previously known and newly identified proteins. A unique knowledge base containing extensive information on the proteins identified in envelope fractions was thus obtained, allowing new insights into this membrane system to be revealed. Altogether, the data we obtained provide unexpected information about plastidial or subplastidial localization of some proteins that were not suspected to be associated to this membrane system. The spectral counting-based strategy was further validated as the compartmentation of well known pathways (for instance, photosynthesis and amino acid, fatty acid, or glycerolipid biosynthesis) within chloroplasts could be dissected. It also allowed revisiting the compartmentation of the chloroplast metabolism and functions.


Migraine and Tension-Type Headache.
Migraine and tension-type headache (TTH) are common primary disorders that carry significant morbidity and socioeconomic effect. In this article, we will review the epidemiology, presentation, and diagnosis of these disorders. First-line acute treatment for migraine consists of analgesics, triptans, and antiemetics, while nonsteroidal anti-inflammatory drugs are the mainstay treatment for TTH. Patients with frequent or chronic headaches warrant prophylactic therapy. For migraine, various classes of preventives can be used (β-blockers, tricyclics, antiepileptics, botulinum toxin), with the choice of therapy tailored to the patient's risk factors and symptoms. For TTH, tricyclics have the most evidence as prophylactic therapy. A new class of medication, monoclonal antibodies to calcitonin gene receptor peptide or its receptor, became available in 2018, and is the first class of medication specifically designed to treat migraine. In addition to pharmacotherapy, we will also review nonpharmacologic interventions as well as neuromodulation for migraine.


Obstetric factors associated with cord blood gas values at birth.
Values of pH, base deficit and Pco2 in 885 unselected samples of umbilical blood collected at birth were related to pregnancy and delivery details. An increase in acidosis in both umbilical vein and artery was associated with placental abruption, maternal pethidine, pre-eclampsia, primigravidity, vaginal operative delivery for fetal distress and a long second stage of labour. Increased acidosis in arterial, but not venous, blood occurred with cord entanglement, vaginal breech delivery, postmaturity, high birthweight and a long first stage of labour. The relationship between obstetric factors and the acid-base state of the neonate suggests a possible role for cord blood gas measurements in assessing the results of obstetric practice.


Cigarette smoke-related oxidants and the development of sub-RPE deposits in an experimental animal model of dry AMD.
Oxidative injury to the retinal pigment epithelium (RPE) has been proposed to be an important injury stimulus relevant to the accumulation of subretinal deposits in age-related macular degeneration (AMD). Cigarette smoking is a major risk factor for AMD, and cigarette smoke-related tar contains high concentrations of a potent oxidant, hydroquinone (HQ). This study was an investigation of the effects of cigarette smoke (CS) and HQ in the development of sub-RPE deposits in an experimental mouse model. Sixteen-month-old C57BL/6 female mice were fed a high-fat diet (HFD) for 4.5 months. Mice were divided into two major experimental groups, one to examine the effects of cigarette smoke and one to study the effects of a defined cigarette smoke component such as HQ. In the first group, mice eyes were exposed to blue-green light (positive controls) or to whole cigarette smoke. A third group with no intervention served as the negative control. In the second experimental group, animals received a purified diet with HQ (0.8%) with low or high fat content for 4.5 months. Mice in both groups were euthanatized at 4.5 months and eyes processed for transmission electron microscopy. As previously demonstrated by our laboratory and others, most mice fed an HFD without other oxidant exposure demonstrated normal morphology or, in a few cases, small nodular basal laminar deposits. Eyes of mice exposed to whole cigarette smoke or to HQ in the food demonstrated a variable degree of basal laminar deposits and diffusely thickened Bruch's membrane. The choriocapillaris endothelium was variably hypertrophic. Exposure to cigarette smoke or the smoke-related redox molecule, HQ, results in the formation of sub-RPE deposits, thickening of Bruch's membrane, and accumulation of deposits within Bruch's membrane. Smoke-related oxidants may be another oxidative injury stimulus to the choriocapillaris and RPE, and may explain the association between cigarette smoking and early AMD.


An innovative off-campus infusion suite designed to improve experiences of patients with cancer.
The Brooklyn Infusion Center of Memorial Sloan-Kettering Cancer Center was established in 2010 to better meet the needs of patients with cancer living in the Brooklyn neighborhood and surrounding areas. A multidisciplinary team comprising clinical, administrative, planning, and other representatives were charged to identify and develop a location that would provide oncology care for patients closer to home and improve the patients' experience. The primary objectives were to provide patient-centered care that accommodates the patients' preference to receive treatment closer to home and to take advantage of technology to establish processes that will provide safe, efficient, convenient, and high-quality care in a cost-effective manner. To achieve these objectives, no laboratory processing or pharmacy services were included in the plan for the Brooklyn location. This allowed the elimination of most of the challenges involved with same-day blood draws and chemotherapy orders. In addition, computer technology is used for teledermatology and other medical visits to maintain the continuity of the patients' care with their multidisciplinary teams at the Manhattan, NY, location. The data presented will illustrate how these processes have improved patients' experiences by reducing wait times for treatment, providing treatment closer to home, and implementing a truly patient-centered nursing care model.


Occupational exposures and symptoms among firefighters and police during the carmel forest fire: the Carmel cohort study.
From 2 to 5 December 2010, Israel experienced the most severe forest fire In its history, resulting in the deaths of rescue workers. Little research exists on the health risks to emergency responders during forest fires, and there is no published research to date on occupational health among firefighters in Israel. To describe the exposures experienced by emer gency responders to smoke, fire retardants and stress; the utilization of protective equipment; and the frequency of corresponding symptoms during and following the Carmel Forest fire. A cohort of 204 firfighers and 68 police who took part in rescue and fire-abating activites during the Carmel Forest fire were recruited from a representative sample of participating stations throughout the country and interviewed regarding their activities during the fire and their coinciding symptoms. Unpaired two-sample t-test compared mean exposures and symptom frequency for firefighters and police. Chi-square estimates of OR and 95%CI are provided for odds of reporting symptoms, incurring injury or being hospitalied for various risk factors Of the study participants, 87% reported having at least one symptom during rescue work at the Carmel Forest fire,with eye irritation (77%) and fatigue (71%) being the most comon. Occupational stress was extremely high during the fire; the average length of time working without rest was 18.4 hours among firefighters. Firefighter and police were exposed to smoke and ocupational stress prolonged periods during the fire. Further research is needed on the residual health effects from exposure to forest fires among emergency responders, and to identify areas for improvement in health preparedness.


Response to ruxolitinib in patients with intermediate-1-, intermediate-2-, and high-risk myelofibrosis: results of the UK ROBUST Trial.
Myelofibrosis is characterized by splenomegaly and debilitating constitutional symptoms that negatively impact patients' quality of life. ROBUST, a UK, open-label, phase II study, evaluated the safety and efficacy of ruxolitinib in patients with myelofibrosis (N = 48), including intermediate-1 risk patients. The primary composite endpoint was the proportion of patients achieving treatment success [≥ 50% reduction in palpable spleen length and/or a ≥ 50% decrease in Myelofibrosis Symptom Assessment Form Total Symptom Score (MF-SAF TSS)] at 48 weeks. This was the first time that efficacy of ruxolitinib in myelofibrosis has been evaluated based on these criteria and the first time the MF-SAF was used in a population of patients solely from the United Kingdom. Overall, 50% of patients and 57% of intermediate-1 risk patients, achieved treatment success; reductions in spleen length and symptoms were observed in all risk groups. The majority of patients (66.7%) experienced ≥ 50% reductions from baseline in spleen length at any time. Improvements in MF-SAF TSS were seen in 80.0%, 72.7%, and 72.2% of intermediate-1, intermediate-2, and high-risk patients, respectively. Consistent with other studies of ruxolitinib, the most common haematological adverse events were anaemia and thrombocytopenia. Results indicate that most patients with myelofibrosis, including intermediate-1 risk patients, may benefit from ruxolitinib treatment.


The impact of comparative effectiveness research on interventional pain management: evolution from Medicare Modernization Act to Patient Protection and Affordable Care Act and the Patient-Centered Outcomes Research Institute.
The Patient-Centered Outcomes Research Institute (PCORI) was established by the Affordable Care Act of 2010 to promote comparative effectiveness research (CER) to assist patients, clinicians, purchasers, and policy-makers in making informed health decisions by advancing the quality and relevance of evidence concerning the manner in which diseases, disorders, and other health conditions can effectively and appropriately be prevented, diagnosed, treated, monitored, and managed through research and evidence synthesis. The development of PCORI is vested in the Medicare Modernization Act (MMA) and the American Recovery and Reinvestment Act (ARRA). The framework of CER and PCORI describes multiple elements which are vested in all 3 regulations including stakeholder involvement, public participation, and open transparent decision-making process. Overall, PCORI is much more elaborate with significant involvement of stakeholders, transparency, public participation, and open decision-making. However, there are multiple issues concerning the operation of such agencies in the United States including the predecessor of Agency for Healthcare Research and Quality (AHRQ), the Agency for Healthcare Policy and Research (AHCPR), AHRQ Effectiveness Health Care programs, and others. The CER in the United States may be described at cross-roads or at the beginnings of a scientific era of CER and evidence-based medicine (EBM). However the United States suffers as other countries, including the United Kingdom with its National Health Services (NHS) and National Institute for Health and Clinical Excellence (NICE), with major misunderstandings of methodology, an inordinate focus on methodological assessment, lack of understanding of the study design (placebo versus active control), lack of involvement of clinicians, and misinterpretation of the evidence which continues to be disseminated. Consequently, PCORI and CER have been described as government-driven solutions without following the principles of EBM with an extensive focus on costs rather than quality. It also has been stated that the central planning which has been described for PCORI and CER, a term devised to be acceptable, will be used by third party payors to override the physician's best medical judgement and patient's best interest. Further, stakeholders in PCORI are not scientists, are not balanced, and will set an agenda with an ultimate problem of comparative effectiveness and PCORI that it is not based on medical science, but rather on political science and not even under congressional authority, leading to unprecedented negative changes to health care. Thus, PCORI is operating in an ad hoc manner that is incompatible with the principles of evidence-based practice.This manuscript describes the framework of PCORI, and the role of CER and its impact on interventional pain management.


A Small-Molecule Inhibitor of trans-Translation Synergistically Interacts with Cathelicidin Antimicrobial Peptides To Impair Survival of Staphylococcus aureus.
Staphylococcus aureus is a leading cause of infection in the United States, and due to the rapid development of resistance, new antibiotics are constantly needed. trans-Translation is a particularly promising antibiotic target because it is conserved in many bacterial species, is critical for bacterial survival, and is unique among prokaryotes. We have investigated the potential of KKL-40, a small-molecule inhibitor of trans-translation, and find that it inhibits both methicillin-susceptible and methicillin-resistant strains of S. aureus KKL-40 is also effective against Gram-positive pathogens, including a vancomycin-resistant strain of Enterococcus faecalis, Bacillus subtilis, and Streptococcus pyogenes, although its performance with Gram-negative pathogens is mixed. KKL-40 synergistically interacts with the human antimicrobial peptide LL-37, a member of the cathelicidin family, to inhibit S. aureus but not other antibiotics tested, including daptomycin, kanamycin, or erythromycin. KKL-40 is not cytotoxic to HeLa cells at concentrations that are 100-fold higher than the effective MIC. We also find that S. aureus develops minimal resistance to KKL-40 even after multiday passage at sublethal concentrations. Therefore, trans-translation inhibitors could be a particularly promising drug target against S. aureus, not only because of their ability to inhibit bacterial growth but also because of their potential to simultaneously render S. aureus more susceptible to host antimicrobial peptides.


Time-dependent activation of the semicarbazide-sensitive amine oxidase (SSAO) from ox lung microsomes.
The activity of ox lung microsomal semicarbazide-sensitive amine oxidase (EC 220.127.116.11; SSAO) towards benzylamine increased 20-fold during incubation at 37 degrees C. After an initial lag-period, activation was first-order with time and complete after approx. 20 h. No significant changes in activity towards methylamine, histamine or 2-phenylethylamine were observed, although mixed-substrate experiments were consistent with the same enzyme being involved in the oxidation of all these substrates, both before and after time-dependent activation. The enzyme-tryptophan fluorescence increased on incubation at 37 degrees C in parallel with the increase in activity towards benzylamine. Treatment of the activated-enzyme preparation with 6 M guanidinium chloride followed by dialysis, caused both the activity towards benzylamine and the fluorescence to fall to that occurring before activation. However, incubation of this preparation at 37 degrees C resulted in increases in fluorescence and activity similar to those seen with the unactivated enzyme. Benzylamine oxidation was inhibited, uncompetitively with respect to oxygen, by high substrate concentrations but no such inhibition was observed with the other amines. Activation resulted in an increase in V(max) for benzylamine oxidation, with no significant alterations in the K(m) or the K(si) for high-substrate inhibition. Kinetic studies were consistent with sequential mechanisms being followed for the oxidation of both benzylamine and methylamine but the dependence on oxygen concentration was complex. These results might indicate that benzylamine follows a different reaction pathway from the other substrates, with substrate-specific activation involving a reaction step that is rate-limiting for benzylamine oxidation but not for the others.


Lifetime stress accelerates epigenetic aging in an urban, African American cohort: relevance of glucocorticoid signaling.
Chronic psychological stress is associated with accelerated aging and increased risk for aging-related diseases, but the underlying molecular mechanisms are unclear. We examined the effect of lifetime stressors on a DNA methylation-based age predictor, epigenetic clock. After controlling for blood cell-type composition and lifestyle parameters, cumulative lifetime stress, but not childhood maltreatment or current stress alone, predicted accelerated epigenetic aging in an urban, African American cohort (n = 392). This effect was primarily driven by personal life stressors, was more pronounced with advancing age, and was blunted in individuals with higher childhood abuse exposure. Hypothesizing that these epigenetic effects could be mediated by glucocorticoid signaling, we found that a high number (n = 85) of epigenetic clock CpG sites were located within glucocorticoid response elements. We further examined the functional effects of glucocorticoids on epigenetic clock CpGs in an independent sample with genome-wide DNA methylation (n = 124) and gene expression data (n = 297) before and after exposure to the glucocorticoid receptor agonist dexamethasone. Dexamethasone induced dynamic changes in methylation in 31.2 % (110/353) of these CpGs and transcription in 81.7 % (139/170) of genes neighboring epigenetic clock CpGs. Disease enrichment analysis of these dexamethasone-regulated genes showed enriched association for aging-related diseases, including coronary artery disease, arteriosclerosis, and leukemias. Cumulative lifetime stress may accelerate epigenetic aging, an effect that could be driven by glucocorticoid-induced epigenetic changes. These findings contribute to our understanding of mechanisms linking chronic stress with accelerated aging and heightened disease risk.


A Novel Polyphenol Conjugate Sensitizes Cisplatin-Resistant Head and Neck Cancer Cells to Cisplatin via Nrf2 Inhibition.
Many cancer cells show acquired resistance to chemotherapeutic agents, such as cisplatin. This is a major cause of cancer treatment failure, and novel agents to overcome resistance are thus urgently required. A novel synthetic polyphenol conjugate, (E)-3-(3,5-dimethoxyphenyl)-1-(2-methoxyphenyl)prop-2-en-1-one (DPP-23), selectively kills tumor cells via the reactive oxygen species (ROS)-mediated unfolded protein response. We investigated the ability of DPP-23 to overcome cisplatin resistance in head and neck cancer (HNC) cells and further clarified its molecular mechanisms of action. Cisplatin-resistant HNC cell lines and their parental and other HNC cell lines were used. The effects of cisplatin and DPP-23 were assessed alone and in combination in HNC and normal cells using cell viability, cell cycle, and cell death assays, by measuring glutathione (GSH), ROS, and protein levels, and via preclinical mouse studies. DPP-23 induced selective cell death in HNC cells, including cisplatin-resistant HNC cells, but spared normal cells, via cellular GSH depletion and ROS accumulation. The effect was blocked by the antioxidant N-acetyl-L-cysteine. DPP-23 activated p53 and its related cell death pathways via a robust accumulation of cellular ROS that involved inhibition of nuclear factor erythroid 2-related factor 2 antioxidant defense mechanisms. Thus, DPP-23 significantly overcame cisplatin resistance in HNC cells in vitro and in vivo As a promising anticancer strategy, ROS generation and subsequent selective cancer cell killing by DPP-23 might help to overcome cisplatin resistance in HNC. Mol Cancer Ther; 15(11); 2620-9. ©2016 AACR.


Effects of nimodipine administration on small bowel mucosa under conditions of laparotomy and consequent 48-hour starvation in a rat model.
The combination of starvation and surgical trauma induces disturbances to the intestinal mucosal structure and function, as well as changes in mucosal barrier function in the rat small bowel. The aim of the present study was to evaluate the effects of nimodipine administration, on intestinal mucosal structural changes and enterocyte apoptosis, following laparotomy and subsequent postsurgical starvation (PSS) in the rat. Thirty Wistar rats were divided into two experimental groups: A: Control group (n=15), where the animal models underwent laparotomy and consequent 48-hours PSS and B: Nimodipine group (n=15), where the rats underwent laparotomy, followed by intraperitoneal nimodipine administration and consequent 48-hour (h) PSS. Small bowel mucosal structural changes and enterocyte epithelial apoptosis were determined 48 h following laparotomy. Nimodipine rats (group B) demonstrated a significant decrease in small bowel villous height in jejunum (p=0.016) and ileum (p=0.002). Similarly, crypt depth decreased in jejunum (p<0.001) and ileum (p<0.001). Nimodipine group exhibited significantly higher apoptotic index in ileum compared to control rats (p=0.006). Nimodipine did not protect the intestinal mucosa from damage caused by surgery and consequent PSS and had obvious damaging effects on intestinal mucosa with derangements to its structure and subsequent mucosal atrophy.


Genetic interactions between UNC-17/VAChT and a novel transmembrane protein in Caenorhabditis elegans.
The unc-17 gene encodes the vesicular acetylcholine transporter (VAChT) in Caenorhabditis elegans. unc-17 reduction-of-function mutants are small, slow growing, and uncoordinated. Several independent unc-17 alleles are associated with a glycine-to-arginine substitution (G347R), which introduces a positive charge in the ninth transmembrane domain (TMD) of UNC-17. To identify proteins that interact with UNC-17/VAChT, we screened for mutations that suppress the uncoordinated phenotype of UNC-17(G347R) mutants. We identified several dominant allele-specific suppressors, including mutations in the sup-1 locus. The sup-1 gene encodes a single-pass transmembrane protein that is expressed in a subset of neurons and in body muscles. Two independent suppressor alleles of sup-1 are associated with a glycine-to-glutamic acid substitution (G84E), resulting in a negative charge in the SUP-1 TMD. A sup-1 null mutant has no obvious deficits in cholinergic neurotransmission and does not suppress unc-17 mutant phenotypes. Bimolecular fluorescence complementation (BiFC) analysis demonstrated close association of SUP-1 and UNC-17 in synapse-rich regions of the cholinergic nervous system, including the nerve ring and dorsal nerve cords. These observations suggest that UNC-17 and SUP-1 are in close proximity at synapses. We propose that electrostatic interactions between the UNC-17(G347R) and SUP-1(G84E) TMDs alter the conformation of the mutant UNC-17 protein, thereby restoring UNC-17 function; this is similar to the interaction between UNC-17/VAChT and synaptobrevin.


Insulin sensing by astrocytes is critical for normal thermogenesis and body temperature regulation.
The important role of astrocytes in the central control of energy balance and glucose homeostasis has recently been recognized. Changes in thermoregulation can lead to metabolic dysregulation, but the role of astrocytes in this process is not yet clear. Therefore, we generated mice congenitally lacking insulin receptors (Ir) in astrocytes (IrKOGFAP mice) to investigate the involvement of astrocyte insulin signaling. IrKOGFAP mice displayed significantly lower energy expenditure and a strikingly lower basal and fasting body temperature. When exposed to cold, however, they were able to mount a thermogenic response. IrKOGFAP mice displayed sex differences in metabolic function and thermogenesis that may contribute to the development of obesity and type II diabetes as early as 2 months of age. While brown adipose tissue exhibited higher adipocyte size in both sexes, more apoptosis was seen in IrKOGFAP males. Less innervation and lower BAR3 expression levels were also observed in IrKOGFAP brown adipose tissue. These effects have not been reported in models of astrocyte Ir deletion in adulthood. In contrast, body weight and glucose regulatory defects phenocopied such models. These findings identify a novel role for astrocyte insulin signaling in the development of normal body temperature control and sympathetic activation of BAT. Targeting insulin signaling in astrocytes has the potential to serve as a novel target for increasing energy expenditure.


HIV surveillance in complex emergencies.
Many studies have shown a positive association between both migration and temporary expatriation and HIV risk. This association is likely to be similar or even more pronounced for forced migrants. In general, HIV transmission in host-migrant or host-forced-migrant interactions depends on the maturity of the HIV epidemic in both the host and the migrant population, the relative seroprevalence of HIV in the host and the migrant population, the prevalence of other sexually transmitted infections (STIs) that may facilitate transmission, and the level of sexual interaction between the two communities. Complex emergencies are the major cause of mass population movement today. In complex emergencies, additional factors such as sexual interaction between forced-migrant populations and the military; sexual violence; increasing commercial sex work; psychological trauma; and disruption of preventive and curative health services may increase the risk for HIV transmission. Despite recent success in preventing HIV infection in stable populations in selected developing countries, internally displaced persons and refugees (or forced migrants) have not been systematically included in HIV surveillance systems, nor consequently in prevention activities. Standard surveillance systems that rely on functioning health services may not provide useful data in many complex emergency settings. Secondary sources can provide some information in these settings. Little attempt has been made, however, to develop innovative HIV surveillance systems in countries affected by complex emergencies. Consequently, data on the HIV epidemic in these countries are scarce and HIV prevention programs are either not implemented or interventions are not effectively targeted. Second generation surveillance methods such as cross-sectional, population-based surveys can provide rapid information on HIV, STIs, and sexual behavior. The risks for stigmatization and breaches of confidentiality must be recognized. Surveillance, however, is a key component of HIV and STI prevention services for forced migrants. It is required to define the high risk groups, target interventions, and ultimately decrease HIV and STI transmission within countries facing complex emergencies. It is also required to facilitate regional control of HIV epidemics.


The White-Knight Hypothesis, or Does the Environment Limit Innovations?
Organisms often harbor latent traits that are byproducts of other adaptations. Such latent traits are not themselves adaptive but can become adaptive in the right environment. Here I discuss several examples of such traits. Their abundance suggests that environmental change rather than new mutations might often limit the origin of evolutionary adaptations and innovations. This is important, because environments can change much faster than new mutations arise. I introduce a conceptual model that distinguishes between mutation-limited and environment-limited trait origins and suggest how experiments could help discriminate between them. Wherever latent traits are plentiful, ecology rather than genetics might determine how fast new adaptations originate and thus how fast adaptive Darwinian evolution proceeds.


Clinical Evaluation of Human Papillomavirus Screening With p16/Ki-67 Dual Stain Triage in a Large Organized Cervical Cancer Screening Program.
As cervical cancer screening transitions from Papanicolaou cytologic screening to primary human papillomavirus (HPV) testing worldwide, effective triage tests are needed to decide who among the HPV-positive women should receive further diagnostic evaluation to avoid unnecessary colposcopies and biopsies. To evaluate the performance of the p16/Ki-67 dual stain (DS) and HPV16/18 genotyping for the triage of HPV-positive women. A prospective observational study was conducted within the cervical cancer screening program at Kaiser Permanente Northern California of 3225 HPV-positive women undergoing HPV and Papanicolaou cytologic testing with a valid DS result from September 16 to October 31, 2015, with follow-up through December 31, 2018. Human papillomavirus screening with partial genotyping and cytologic triage compared with DS triage. Cervical intraepithelial neoplasia grade 3 or more severe (CIN3+) and grade 2 or more severe (CIN2+), diagnosed within 3 years after sample collection. A total of 3225 women (mean [SD] age, 37.9 [11.3] years) participated in the study. For triage of HPV-positive women with partial genotyping, DS showed better risk stratification for CIN3+ than did Papanicolaou cytologic testing, with women with positive DS results having a higher risk than women with positive Papanicolaou test results for CIN3+ (218 of 1818 [12.0%; 95% CI, 10.5%-13.5%] vs 219 of 2128 [10.3%; 95% CI, 9.0%-11.6%]; P = .005). Similarly, DS showed better risk stratification for CIN3+ compared with Papanicolaou cytologic testing in HPV-positive women, irrespective of genotyping. The greatest reassurance against CIN3+ was observed in HPV16/18-negative women with negative DS results, with a risk low enough to extend retesting intervals. Dual stain triage strategies required substantially fewer colposcopies per detection of CIN3+ compared with Papanicolaou cytologic testing, with a 32.1% (859 of 2677) reduction of colposcopies compared with the currently recommended triage strategy of HPV screening with Papanicolaou cytologic testing. Results for CIN2+ were very similar. Triage of HPV-positive women with DS was superior to Papanicolaou cytologic testing in this study, demonstrating equal immediate detection of precancerous lesions and substantially reduced referral to colposcopy. These findings suggest that DS can safely replace Papanicolaou cytologic testing as a triage strategy for primary HPV screening, and that retesting intervals in HPV16/18-negative women with negative DS results can be safely extended to 3 years.


Formulation and evaluation of floating tablets of liquorice extract.
Floating tablets prolong the gastric residence time of drugs, improve bioavailability, and facilitate local drug delivery to the stomach. With this objective, floating tablets containing aqueous extract of liquorice as drug was prepared for the treatment of Helicobacter pylori and gastric ulcers. The aqueous extract of liquorice was standardized by HPTLC. Tablets containing HPMC K100M (hydrophilic polymer), liquorice extract, sodium bicarbonate (gas generating agent), talc, and magnesium stearate were prepared using direct compression method. The formulations were evaluated for physical parameters like diameter, thickness, hardness, friability, uniformity of weight, drug content, buoyancy time, dissolution, and drug release mechanism. The formulations were optimized on the basis of buoyancy time and in vitro drug release. The diameter of all formulations was in the range 11.166-11.933 mm; thickness was in the range 4.02-4.086 mm. The hardness ranged from 3.1 to 3.5 kg/cm(2). All formulations passed the USP requirements for friability and uniformity of weight. The buoyancy time of all tablet formulations was less than 5 min and tablet remained in floating condition throughout the study. All the tablet formulations followed zero-order kinetics and Korsemeyer-Peppas model in drug release. The optimized formulation was found to be F6 which released 98.3% of drug in 8 h in vitro, while the buoyancy time was 3.5 min. Formulations containing psyllium husk, sodium bicarbonate and HPMC K100M in combination can be a promising for gastroretentive drug delivery systems.


The Landscape of SNCA Transcripts Across Synucleinopathies: New Insights From Long Reads Sequencing Analysis.
Dysregulation of alpha-synuclein expression has been implicated in the pathogenesis of synucleinopathies, in particular Parkinson's Disease (PD) and Dementia with Lewy bodies (DLB). Previous studies have shown that the alternatively spliced isoforms of the SNCA gene are differentially expressed in different parts of the brain for PD and DLB patients. Similarly, SNCA isoforms with skipped exons can have a functional impact on the protein domains. The large intronic region of the SNCA gene was also shown to harbor structural variants that affect transcriptional levels. Here, we apply the first study of using long read sequencing with targeted capture of both the gDNA and cDNA of the SNCA gene in brain tissues of PD, DLB, and control samples using the PacBio Sequel system. The targeted full-length cDNA (Iso-Seq) data confirmed complex usage of known alternative start sites and variable 3' UTR lengths, as well as novel 5' starts and 3' ends not previously described. The targeted gDNA data allowed phasing of up to 81% of the ~114 kb SNCA region, with the longest phased block exceeding 54 kb. We demonstrate that long gDNA and cDNA reads have the potential to reveal long-range information not previously accessible using traditional sequencing methods. This approach has a potential impact in studying disease risk genes such as SNCA, providing new insights into the genetic etiologies, including perturbations to the landscape the gene transcripts, of human complex diseases such as synucleinopathies.


Changes in the clinical and epidemiological features of group A streptococcal bacteraemia in Australia's Northern Territory.
Invasive group A streptococcus (iGAS) disease is an important cause of mortality globally. The incidence of iGAS in Australia's tropical Northern Territory (NT) has been previously reported as 32.2/100 000 in Indigenous people for the period 1991-1996. We aimed to measure the incidence and severity of iGAS disease in the NT since this time. We collected demographic data for all GAS blood culture isolates over a 12-year period (1998-2009) from the three hospital laboratories serving the tropical NT. We then collected detailed clinical information from hospital records and databases for the subset of these patients who were admitted to Royal Darwin Hospital during 2005-2009. There were 295 confirmed cases of GAS bacteraemia over the study period, with a mean (SD) age of 42.1 (22.0) years, and 163 (55.0%) were male. The annual age-adjusted incidence was 15.2 (95% CI 13.4-16.9)/100 000 overall and 59.4 (95% CI 51.2-67.6) in Indigenous Australians. For 2005-2009, there were 123 cases with the most common focus of infection being skin/soft tissue [44 (35.6%)]; 29 patients (23.6%) required intensive care unit admission and 20 (16.3%) had streptococcal toxic shock syndrome. Antecedent sore throat or use of non-steroidal anti-inflammatory drugs was rare, but current or recent scabies, pyoderma and trauma were common. The incidence and severity of iGAS are high and increasing in tropical northern Australia, and urgent attention is needed to improve surveillance and the social determinants of health in this population. This study adds to emerging data suggesting increasing importance of iGAS in low- and middle-income settings globally.


Digital pathology: current status and future perspectives.
During the last decade pathology has benefited from the rapid progress of image digitizing technology. The improvement in this technology had led to the creation of slide scanners which are able to produce whole slide images (WSI) which can be explored by image viewers in a way comparable to the conventional microscope. The file size of the WSI ranges from a few megabytes to several gigabytes, leading to challenges in the area of image storage and management when they will be used routinely in daily clinical practice. Digital slides are used in pathology for education, diagnostic purposes (clinicopathological meetings, consultations, revisions, slide panels and, increasingly, for upfront clinical diagnostics) and archiving. As an alternative to conventional slides, WSI are generally well accepted, especially in education, where they are available to a large number of students with the full possibilities of annotations without the problem of variation between serial sections. Image processing techniques can also be applied to WSI, providing pathologists with tools assisting in the diagnosis-making process. This paper will highlight the current status of digital pathology applications and its impact on the field of pathology.


Survey of cannabis use in patients with amyotrophic lateral sclerosis.
Cannabis (marijuana) has been proposed as treatment for a widening spectrum of medical conditions and has many properties that may be applicable to the management of amyotrophic lateral sclerosis (ALS). This study is the first, anonymous survey of persons with ALS regarding the use of cannabis. There were 131 respondents, 13 of whom reported using cannabis in the last 12 months. Although the small number of people with ALS that reported using cannabis limits the interpretation of the survey findings, the results indicate that cannabis may be moderately effective at reducing symptoms of appetite loss, depression, pain, spasticity, and drooling. Cannabis was reported ineffective in reducing difficulties with speech and swallowing, and sexual dysfunction. The longest relief was reported for depression (approximately two to three hours).


Treatment of temporohyoid osteoarthropathy in horses with a basihyoid-ceratohyoid disarticulation technique: 6 cases (2018-2019).
To describe a technique for basihyoid-ceratohyoid disarticulation (BCD) in standing sedated horses affected by temporohyoid osteoarthropathy (THO) and report outcomes for horses that underwent the procedure. 6 client-owned horses. Electronic medical records of a veterinary teaching hospital were searched to identify horses that underwent BCD for treatment of THO from 2018 to 2019. Signalment, clinical data, use of the horse, and complications were recorded. Follow-up data obtained by telephone interview with owners included the clinical outcome and time to improvement after surgery, any persistent clinical signs, horse's activity level before onset of clinical signs and after BCD, subsequent use of the horse, and whether they would pursue the same treatment again. All horses tolerated the procedure well, with no complications and improved neurologic function after BCD. Five of 6 horses had a reported activity level equal to or greater than that prior to having signs of THO. Three of 3 horses with acute ataxia prior to BCD reportedly had full resolution of this sign; 3 of 4 horses with facial nerve deficits prior to BCD had mild residual facial nerve deficits at follow-up. All owners indicated they would pursue BCD again. The BCD procedure was performed safely in this sample of THO-affected horses that were sedated while standing, avoiding risks associated with general anesthesia and resulting in no adverse effects such as iatrogenic injury to neurovascular structures.


Photopolymerization-induced crystallization and phase separation in poly(ethylene oxide)/triacrylate blends.
The present article describes experimental and theoretical investigations of miscibility and crystallization behavior of blends of poly(ethylene oxide) (PEO) and triacrylate monomer (TA) using differential scanning calorimetry and optical microscopy. The PEO/TA blends manifested a single T(g) varying systematically with composition suggestive of a miscible character in their amorphous states. Moreover, there occurs melting point depression of PEO crystals with increasing TA. A phase diagram was subsequently established that exhibited a solid+liquid coexistence region bound by the liquidus and solidus lines, followed by an upper critical solution temperature (UCST) at a lower temperature. The emerging phase morphology was investigated to verify the coexistence regions. Upon photopolymerization in the isotropic melt above the melting point depression curve, both the UCST and the melting temperatures move upward and eventually surpass the reaction temperature, resulting in phase separation as well as crystallization of PEO driven by the changing supercooling, i.e., the thermodynamic driving force. Of particular interest is the interplay between photopolymerization-induced phase separation and crystallization, which eventually determines the final phase morphology of the PEO/TA blend such as crystalline lamellae, sheaf, or spherulites in isotropic liquid, phase separated domains, and viscous fingering liquids.


Impact of fresh or used litter on the posthatch immune system of commercial broilers.
This study was carried out to investigate the effects of exposure of growing broiler chickens of commercial origin to used poultry litter on intestinal and systemic immune responses. The litter types evaluated were fresh wood shavings or used litter obtained from commercial poultry farms with or without a history of gangrenous dermatitis (GD). Immune parameters measured were serum nitric oxide (NO) levels, serum antibody titers against Eimeria or Clostridium perfringens, mitogen-induced spleen cell proliferation, and intestinal intraepithelial lymphocyte or splenic lymphocyte subpopulations. At 43 days posthatch, birds raised on used litter from a GD farm had higher serum NO levels and greater Eimeria or C. perfringens antibody levels compared with chickens raised on fresh litter or used, non-GD litter. Birds raised on non-GD and GD used litter had greater spleen cell mitogenic responses compared with chickens raised on fresh litter. Finally, spleen and intestinal lymphocyte subpopulations were increased or decreased depending on the litter type and the surface marker analyzed. Although it is likely that the presence of Eimeria oocysts and endemic viruses varies qualitatively and quantitatively between flocks and, by extension, varies between different used litter types, we believe that these data provide evidence that exposure of growing chicks to used poultry litter stimulates humoral and cell-mediated immune responses, presumably due to contact with contaminating enteric pathogens.


Free will and the obesity epidemic.
The increase in body weight in the USA over the past several decades is now commonly referred to as the 'obesity epidemic'. An empirical analysis of the literature suggests that the increased weight can be accounted for by an increase in food intake. The solution to the obesity epidemic, therefore, must centre on a reduction in food consumption, a position well accepted by the American population who think that they, as individuals, are responsible for their adiposity by holding the belief that the decision as to what and how much to eat is determined by their own free will. The evidence demonstrates, however, that this is not true. Variables such as portion size, variety of foods offered, fat content of the diet, the number of people eating, the location where eating occurs and even watching food advertisements act as 'food primes' causing individuals to increase their energy intake. Despite the plethora of diets, weight-loss clubs, drugs and mechanical devices available to facilitate weight loss, once treatment is terminated and people return to the 'free' environment, their weight returns to pre-treatment levels. Only when individuals are protected from environmental variables by gastric surgery or limited to consume only portion-controlled meals can they successfully maintain a reduced weight. Combining the technique of daily weight monitoring with accepting that our eating behaviour is not determined totally by our free choice, we may be able to curb the obesity epidemic.


Long-term ethanol effects on acute stress responses: modulation by dynorphin.
The brain stress-response system is critically involved in the addiction process, stimulating drug consumption and the relapse to drug taking in abstinent addicts. At the same time, its functioning is affected by chronic drug exposure. Here, we have investigated the role of the endogenous opioid peptide dynorphin as a modulator of effects of long-term ethanol consumption on the brain stress-response system. Using the two-bottle choice paradigm, we demonstrate an enhanced ethanol preference in male dynorphin knockout mice. Exposure to mild foot shock increased ethanol consumption in wild-type control littermates, but not in dynorphin-deficient animals. Blood adrenocorticotropic hormone levels determined 5 minutes after the shock were not affected by the genotype. We also determined the neuronal reactivity after foot shock exposure using c-Fos immunoreactivity in limbic structures. This was strongly influenced by both genotype and chronic ethanol consumption. Long-term alcohol exposure elevated the foot shock-induced c-Fos expression in the basolateral amygdala in wild-type animals, but had the opposite effect in dynorphin-deficient mice. An altered c-Fos reactivity was also found in the periventricular nucleus, the thalamus and the hippocampus of dynorphin knockouts. Together these data suggest that dynorphin plays an important role in the modulation of the brain stress-response systems after chronic ethanol exposure.


Linking snake habitat use to nest predation risk in grassland birds: the dangers of shrub cover.
Extremes in rangeland management, varying from too-frequent fire and intensive grazing to the suppression of both, threaten rangeland ecosystems worldwide. Intensive fire and grazing denude and homogenize vegetation whereas their suppression increases woody cover. Although habitat loss is implicated in grassland bird declines, degradation through intensive management or neglect also decreases breeding habitat and may reduce nesting success through increased rates of nest predation. Snakes are important nest predators, but little is known about how habitat use in snakes relates to predation risk for grassland birds nesting within tallgrass prairie subjected to different grazing and fire frequencies. We evaluated nest survival in the context of habitat used by nesting songbirds and two bird-eating snakes, the eastern yellowbelly racer Coluber constrictor flaviventris and Great Plains ratsnake Pantherophis emoryi. Daily nest survival rates decreased with increasing shrub cover and decreasing vegetation height, which characterize grasslands that have been neglected or intensively managed, respectively. Discriminant function analysis revealed that snake habitats were characterized by higher shrub cover, whereas successful nests were more likely to occur in areas with tall grass and forbs but reduced shrub cover. Because snakes often use shrub habitat, birds nesting in areas with increased shrub cover may be at higher risk of nest predation by snakes in addition to other predators known to use shrub habitat (e.g., mid-sized carnivores and avian predators). Depredated nests also occurred outside the discriminant space of the snakes, indicating that other predators (e.g., ground squirrels Spermophilus spp. and bullsnakes Pituophis catenifer) may be important in areas with denuded cover. Targeted removal of shrubs may increase nest success by minimizing the activity of nest predators attracted to shrub cover.


Determination of optical parameters and thickness of weakly absorbing thin films from reflectance and transmittance spectra.
A method for determining the optical constants and the thickness of weakly absorbing thin films on substrates is proposed. In this method only the reflectance and transmittance spectra obtained at a single arbitrary angle of incidence are used, provided that the former reveals several interference extrema. The calculation procedure is based on relatively simple relations suitable for the programmed realization and does not call for the prescription of the initial values of the parameters to be determined. The method proposed is fairly accurate and allows one to uniquely solve the inverse problem of spectrophotometry. The optical constants and the thickness of an As(x)Se(y) film formed on a glass substrate have been determined by the proposed method in the visible region of the spectrum.


Time-spatial drift of decelerating electromagnetic pulses.
A time dependent electromagnetic pulse generated by a current running laterally to the direction of the pulse propagation is considered in paraxial approximation. It is shown that the pulse envelope moves in the time-spatial coordinates on the surface of a parabolic cylinder for the Airy pulse and a hyperbolic cylinder for the Gaussian. These pulses propagate in time with deceleration along the dominant propagation direction and drift uniformly in the lateral direction. The Airy pulse stops at infinity while the asymptotic velocity of the Gaussian is nonzero.


A pilot study of vitamins to lower plasma homocysteine levels in Alzheimer disease.
Authors determined the impact of high-dose vitamin supplements on plasma homocysteine levels in patients with Alzheimer disease (AD). Authors used an open-label trial of folic acid, vitamin B(12), and vitamin B(6), in combination for 8 weeks, with measurement of plasma homocysteine levels in the fasting state and after methionine-loading. A total of 69 subjects with AD were enrolled, including 33 who were taking standard multivitamin supplements; 66 were available at 8-week follow-up. The high-dose vitamin regimen was associated with a significant reduction in fasting and post-methionine-loading homocysteine. Reductions were greater in the subgroup not using multivitamins, but were also significant in the multivitamin users. High-dose vitamin supplementation reduces homocysteine levels in patients with AD. The effect of supplementation on rate of cognitive decline will be assessed later in a randomized, double-blind study.


A STUDY OF THE FINE STRUCTURE OF THE EPIDERMIS OF RANA PIPIENS.
The epidermis of adult Rana pipiens has been studied by electron microscopy and histological and histochemical methods. It was found that the epidermis is engaged in the production of both keratin and mucus. The basal cells are mainly filled with tonofilaments, whereas the cells located in the mid-portion of the epidermis contain both tonofilaments and mucous granules. Golgi vesicles and endoplasmic reticulum are found in relative abundance in the mucus-producing cells and seem to be involved in the production of mucous granules. The mucus seen was partly retained within the cells and partly secreted into the intercellular spaces. The outermost keratinized cells contain mainly filaments and a few remnants of cell constituents.