Patients with hypothermic sepsis have a unique gene expression profile compared to patients with fever and sepsis.
The pathophysiology of hypothermia during sepsis is unclear. Using genomic profiling of blood leukocytes, we aimed to determine if hypothermia is associated with a different gene expression profile compared to fever during sepsis. Patients with sepsis and either hypothermia or fever within 24 hours after ICU admission were included in the study (n = 168). Hypothermia was defined as body temperature below 36 °C. Fever was defined as body temperature equal to or above 38.3°C. We compared blood gene expression (whole-genome transcriptome in leukocytes) in hypothermic septic compared to febrile septic patients in an unmatched analysis and matched for APACHE IV score and the presence of shock. In total, 67 septic patients were hypothermic and 101 patients were febrile. Hypothermia was associated with a distinct gene expression profile in both unmatched and matched analyses. There were significant differences related to the up- and downregulation of canonical signalling pathways. In the matched analysis, the top upregulated gene was cold-inducible mRNA binding protein (CIRBP) which plays a role in cold-induced suppression of cell proliferation. In addition, we found three signalling pathways significantly upregulated in hypothermic patients compared to febrile patients; tryptophan degradation X, phenylalanine degradation IV and putrescine degradation III. In conclusion, there are distinct signalling pathways and genes associated with hypothermia, including tryptophan degradation and CIRBP expression, providing a possible link to the modulation of body temperature and early immunosuppression. Future studies may focus on the canonical signalling pathways presented in this paper to further investigate spontaneous hypothermia in sepsis.


De novo primary squamous cell carcinoma of the ovary: a case of a rare malignancy with an aggressive clinical course.
Ovarian squamous cell carcinoma is a rare malignancy and the occurrence is attributable to malignant transformation of an existing ovarian dermoid cyst. The de novo occurrence of squamous cell carcinoma of the ovary, in the absence of an antecedent ovarian dermoid, is extremely rare. The case of a 31 year old Asian woman, evaluated for abdominal distension and discomfort is presented. Abdominal CT was suggestive of a malignant neoplastic process. Laparotomy confirmed a malignant tumour with involvement of the right adnexa and extension into the omentum and bowel. Surgical debulking, hysterectomy, bilateral salpingo-ophorectomy and total omentectomy and bowel resection was performed. Histopathology demonstrated squamous cell carcinoma arising from the right ovary with no co-existing ovarian dermoid. The postoperative period was significant for disease progression despite adjuvant chemotherapy.


Calcium regulates the commitment of murine erythroleukemia cells to terminal erythroid differentiation.
An alteration in the rate of calcium transport appears to be the rate-limiting event for the commitment of murine erythroleukemia (MEL) cells to initiate a program of terminal erythroid differentiation. The dimethyl sulfoxide (DMSO)-induced commitment of MEL cells to erythroid differentiation can be inhibited by treatment of cells with the calcium-chelating agent EGTA. Upon removal of EGTA, cells initiate commitment without the 12-h lag normally observed after treatment with DMSO alone. Treatment of cells with DMSO in the presence of calcium ionophore A23187 causes cells to initiate commitment from time zero with no lag. These results suggest that the lag is the time required for DMSO to alter the calcium transport properties of the cell.


A single-step procedure of recombinant library construction for the selection of efficiently produced llama VH binders directed against cancer markers.
Heavy chain antibodies are naturally occurring in camelidae (camels and llamas). Their variable domain (VHH) can be efficiently produced as a recombinant protein in E. coli with a large range of applications in the fields of diagnostics and immunotherapy. Standard cloning approach involves resolution of VHH from the heavy chain variable domain of conventional antibodies (VH) by a nested PCR amplification followed by a phage display based selection. Present work illustrates that in contrast to usual finding, specific, good affinity and efficiently expressed VH domain of conventional antibodies can be selected from the co-amplification products of VH and VHH cDNAs. Sequence analysis illustrated that following the two first rounds of selection against cancer markers, similar number of VH and VHH binders were observed. However, after a third round, the more specific binders directed against p53, VEGF, BCL-2 proteins surprisingly contain only VH specific hallmarks. Characterisation of the specificity, affinity and productivity of selected VH binders is described. Because llama VHs show higher sequence and structural homology with the human VH III group than llama VHHs (Vu et al., 1997), they constitute very interesting agents in therapeutic applications, especially in human immunotherapy and cancer treatment.


Sinorhizobium meliloti flavin secretion and bacteria-host interaction: role of the bifunctional RibBA protein.
Sinorhizobium meliloti, the nitrogen-fixing bacterial symbiont of Medicago spp. and other legumes, secretes a considerable amount of riboflavin. This precursor of the cofactors flavin mononucleotide and flavin adenine dinucleotide is a bioactive molecule that has a beneficial effect on plant growth. The ribBA gene of S. meliloti codes for a putative bifunctional enzyme with dihydroxybutanone phosphate synthase and guanosine triphosphate (GTP) cyclohydrolase II activities, catalyzing the initial steps of the riboflavin biosynthesis pathway. We show here that an in-frame deletion of ribBA does not cause riboflavin auxotrophy or affect the ability of S. meliloti to establish an effective symbiosis with the host plant but does affect the ability of the bacteria to secrete flavins, colonize host-plant roots, and compete for nodulation. A strain missing the RibBA protein retains considerable GTP cyclohydrolase II activity. Based on these results, we hypothesize that S. meliloti has two partly interchangeable modules for biosynthesis of riboflavin, one fulfilling the internal need for flavins in bacterial metabolism and the other producing riboflavin for secretion. Our data also indicate that bacteria-derived flavins play a role in communication between rhizobia and the legume host and that the RibBA protein is important in this communication process even though it is not essential for riboflavin biosynthesis and symbiosis.


Corneal Guttae After Descemet Membrane Endothelial Keratoplasty.
The aim of this study was to report on the occurrence of corneal guttae after Descemet membrane endothelial keratoplasty (DMEK). In this retrospective case series, 13 eyes of 13 patients who underwent DMEK at 2 tertiary referral centers between 2007 and 2021 (average available follow-up 73 ± 52 months, range 18-174 months) and showed corneal guttae during postoperative examinations were included. Eye bank images were retrospectively reviewed. Occurrence of guttae was observed by specular microscopy in 13 eyes. In 11 cases, presence of guttae was confirmed by confocal microscopy and in 1 case by histology. Five eyes showed an increase in guttae density during the postoperative course. Surgery indications were Fuchs endothelial corneal dystrophy (n = 11), pseudophakic bullous keratopathy (n = 1), and DMEK graft failure after allograft rejection (n = 1); the latter eye had shown no signs of guttae after primary DMEK. Two eyes with guttae required a repeat DMEK due to graft failure. At the last available follow-up, all 11 remaining eyes had clear corneas and 10 eyes had a best-corrected visual acuity of ≥0.9 (decimal). During donor cornea processing in the eye bank, no guttae were observed on the donor tissue. Corneal guttae can occur after DMEK including in eyes operated for indications other than Fuchs endothelial corneal dystrophy and most likely guttae were present on the donor graft but were not detectable by routine slit-lamp and light microscopy evaluation in the eye bank. Postoperative guttae density varies among patients and especially small isolated guttae do not seem to affect clinical outcomes.


Effects of dietary fat levels and feeding durations on musculoskeletal health in female rats.
This study evaluates the effects of dietary fat levels and feeding durations on body composition, bone properties, and endocrine factors in female rats. Forty-eight 3-month-old female rats were assigned into a 2 (low-fat diet vs. high-fat diet) × 2 (4-month vs. 8-month feeding duration) factorial design. The body composition, bone matrix and microstructure, and endocrine factors were examined. Compared to the low-fat diet, the high-fat diet (i) increased the % fat mass, femoral mineral density, trabecular and cortical formation and erosion, osteoblast and osteoclast numbers, insulin-like growth factor-I, and leptin and (ii) decreased the % fat-free mass, bone strength, and trabecular and cortical bone volume and thickness. Relative to the 4-month feeding duration, the 8-month feeding duration (i) increased the femoral mineral density, trabecular separation, formation and erosion, endocortical formation rate, and osteoclast number and (ii) decreased the trabecular bone volume, thickness, and number, osteoblast number, and adiponectin. Interactions between dietary fat level and feeding duration were observed in bone strength, trabecular thickness and formation rate, as well as cortical bone volume and mineral apposition rate. Both dietary fat level and feeding duration affect the bone mass and microstructure in female rats, possibly through the modulation of body composition and endocrine factors.


Helicobacter pylori CagA triggers expression of the bactericidal lectin REG3γ via gastric STAT3 activation.
Most of what is known about the Helicobacter pylori (H. pylori) cytotoxin, CagA, pertains to a much-vaunted role as a determinant of gastric inflammation and cancer. Little attention has been devoted to potential roles of CagA in the majority of H. pylori infected individuals not showing oncogenic progression, particularly in relation to host tolerance. Regenerating islet-derived (REG)3γ encodes a secreted C-type lectin that exerts direct bactericidal activity against Gram-positive bacteria in the intestine. Here, we extend this paradigm of lectin-mediated innate immunity, showing that REG3γ expression is triggered by CagA in the H. pylori-infected stomach. In human gastric mucosal tissues, REG3γ expression was significantly increased in CagA-positive, compared to CagA-negative H. pylori infected individuals. Using transfected CagA-inducible gastric MKN28 cells, we recapitulated REG3γ induction in vitro, also showing that tyrosine phosphorylated, not unphosphorylated CagA triggers REG3γ transcription. In concert with induced REG3γ, pro-inflammatory signalling downstream of the gp130 cytokine co-receptor via the signal transducer and activator of transcription (STAT)3 and transcription of two cognate ligands, interleukin(IL)-11 and IL-6, were significantly increased. Exogenous IL-11, but not IL-6, directly stimulated STAT3 activation and REG3γ transcription. STAT3 siRNA knockdown or IL-11 receptor blockade respectively abrogated or subdued CagA-dependent REG3γ mRNA induction, thus demonstrating a requirement for uncompromised signalling via the IL-11/STAT3 pathway. Inhibition of the gp130-related SHP2-(Ras)-ERK pathway did not affect CagA-dependent REG3γ induction, but strengthened STAT3 activation as well as augmenting transcription of mucosal innate immune regulators, IL-6, IL-8 and interferon-response factor (IRF)1. Our results support a model of CagA-directed REG3γ expression in gastric epithelial cells via activation of the IL-11/gp130/STAT3 pathway. This response might allow Gram-negative H. pylori to manipulate host immunity to favour its own survival, by reducing the fitness of co-habiting Gram-positive bacteria with which it competes for resources in the gastric mucosal niche.


Expression of intestinal MUC17 membrane-bound mucin in inflammatory and neoplastic diseases of the colon.
To determine the cellular location and expression of MUC17 mucin in specimens of normal, inflamed and neoplastic colon. Immunohistochemical analysis of human surgical resection specimens (n=106) was performed with a specific antibody to the MUC17 apomucin protein. A semi-quantitative scoring system was used to measure MUC17 expression. In various colon cancer cell lines, the MUC17 expression was examined by immunoblot analysis and normal RT-PCR. MUC17 was highly expressed on the surface epithelium and crypts of colonic mucosa. In contrast, the expression of MUC17 was significantly decreased in colonic mucosa of chronic ulcerative colitis (p<0.0001) and ischaemic colitis (p=0.003). Similarly, MUC17 expression was decreased in hyperplastic polyps (p=0.0003), tubular and tubulovillous adenomas (p<0.0001) and colon cancers (p<0.0001). Furthermore, of eight different colon cancer cell lines, MUC17 expression was only detected in LS174T and LS180 cells. Results indicate that the potential protective effects of this membrane-bound mucin are primarily or secondarily diminished in inflammatory and neoplastic conditions. Further research is needed to determine the specific role of MUC17 in the pathogenesis of these conditions.


Choline supplementation alleviates fluoride-induced testicular toxicity by restoring the NGF and MEK expression in mice.
Fluoride is known to cause male reproductive toxicity, and the elucidation of its underlying mechanisms is an ongoing research focus in reproductive toxicology and epidemiology. Choline, an essential nutrient, has been extensively studied for its benefits in nervous system yet was rarely discussed for its prospective effect in male reproductive system. This study aims to explore the potential protective role of choline against NaF-induced male reproductive toxicity via MAPK pathway. The male mice were administrated by 150mg/L NaF in drinking water, 5.75g/kg choline in diet, and their combination respectively from maternal gestation to postnatal 15weeks. The results showed that fluoride exposure reduced body weight growth, lowered sperm count and survival percentages, altered testicular histology, down-regulated the mRNA expressions of NGF, Ras, Raf, and MEK genes in testes, as well as significantly decreased the expressions of both NGF and phosphor-MEK proteins in testes. Examination of data from choline-treated mice revealed that choline supplementation ameliorated these fluoride-induced changes. Taken together, our findings suggest that choline supplementation alleviates fluoride-induced testicular toxicity by restoring the NGF and phosphor-MEK expression. The suitable dosage and supplementation periods of choline await further exploration.


Autonomous effects of shear stress and cyclic circumferential stretch regarding endothelial dysfunction and oxidative stress: an ex vivo arterial model.
Cyclic circumferential stretch and shear stress caused by pulsatile blood flow work in concert, yet are very different stimuli capable of independently mediating endothelial function by modulating eNOS expression, oxidative stress (via production of superoxide anion) and NO bioavailability. Porcine carotid arteries were perfused using an ex vivo arterial support system for 72 h. Groups we created by combining normal (5%) and reduced (1%) stretch with high shear (6 +/- 3 dynes/cm(2)) and oscillatory shear (0.3 +/- 3 dynes/cm(2)) stress while maintaining a pulse pressure of 80 +/- 10 mm Hg. Oscillatory flow and reduced stretch both proved detrimental to endothelial function, whereas oscillatory flow alone dominated total endogenous vascular wall superoxide anion production. Yet, when superoxide anion production was analyzed in just the endothelial region, we observed that it was modulated more significantly by reduced cyclic stretch than by oscillatory shear, emphasizing an important distinction between shear- and stretch-mediated effects to the vascular wall. Western blotting analysis of eNOS and nitrotyrosine proved that they too are more significantly negatively modulated by oscillatory flow than by reduced stretch. These findings point out how shear and stretch stimulate regions of the vascular wall differently, affecting NO bioavailability and contributing to vascular disease.


Organotin Release from Polyvinyl Chloride Microplastics and Concurrent Photodegradation in Water: Impacts from Salinity, Dissolved Organic Matter, and Light Exposure.
Photochemical weathering leads to degradation of microplastics and releases chemical additives, polymeric fragments, and/or byproducts. This study evaluated the release kinetics of organotin compounds (OTCs) from three different sized (10-300 μm) polyvinyl chloride (PVC) microplastics under UV- and visible light irradiation. Four OTCs, dimethyltin (DMT), monomethyltin (MMT), dibutyltin (DBT), and monobutyltin (MBT), were found to release from PVC particles after 24 h leaching in darkness ranging from 2 to 20 μg·g-PVC-1. Under UV/visible light irradiation, only DMT and DBT were detectable, whereas MMT and MBT were not detected due to rapid photodegradation. The total tin concentrations (including organic and inorganic tins) in the aqueous phase monotonically increased under light exposure. By contrast, they reached plateaus after 24 h in darkness, confirming the photodegradation of OTCs. A release kinetics model was established and correctly interpreted the microplastics size effect on the OTC release process. Finally, the impacts of salinity and dissolved organic matter (DOM) were investigated. The release and photodegradation of OTCs were both inhibited at high salinity conditions, probably due to the enhanced readsorption of OTCs on PVC microplastics and the formation of halogen radicals that were less reactive toward neutral OTCs. The presence of DOM, however, increased OTCs release probably because the excited state triplet DOM (3DOM*) formed and reacted with OTCs from PVC microplastics.


The power of touch: external applications from whole medical systems in the care of cancer patients (literature review).
In the context of whole medical systems (WMSs), external applications are manual treatments that stem from complementary medicine and are performed by practitioners, nurses and caregivers through the application of interpersonal touch. These applications work via physiological, psychological and social processes and are intended for holistic healing. In integrative oncology, these applications are used to improve patients' wellbeing. The purpose of the review is to collect and compare research results concerning external applications from WMSs applied to patients with cancer. We conducted a literature review to explore the effects of external applications on patients undergoing cancer treatment. Inclusion criteria were external applications from Anthroposophic Medicine, Traditional Chinese Medicine (TCM) and Ayurvedic Medicine, examined detached from the effect interdependencies of their respective WMSs. Searches identified 111 publications. Final inclusions were four Shiatsu studies (from TCM), supplemented by two case reports exploring anthroposophic external applications. The literature review suggests that external applications may improve patients' wellbeing during cancer treatment and are associated with physical and psychological effects that potentially enhance supportive cancer care. The number and quality of published studies in the field is limited. Future empirical investigations should assess the effectiveness and underlying effects in patients with cancer, adhere to a patient-tailored approach, and investigate the systemic complexity of touch-based applications from WMSs.


Vaginal microbiota and mucosal pharmacokinetics of tenofovir in healthy women using tenofovir and tenofovir/levonorgestrel vaginal rings.
Recent data support that the vaginal microbiota may alter mucosal pharmacokinetics (PK) of topically delivered microbicides. Our team developed an intravaginal ring (IVR) that delivers tenofovir (TFV) (8-10 mg/day) alone or with levonorgestrel (LNG) (20 ug/day). We evaluated the effect of IVRs on the vaginal microbiota, and describe how the vaginal microbiota impacts mucosal PK of TFV. CONRAD A13-128 was a randomized, placebo controlled phase I study. We randomized 51 women to TFV, TFV/LNG or placebo IVR. We assessed the vaginal microbiota by sequencing the V3-V4 regions of 16S rRNA genes prior to IVR insertion and after approximately 15 days of use. We measured the concentration of TFV in the cervicovaginal (CV) aspirate, and TFV and TFV-diphosphate (TFV-DP) in vaginal tissue at the end of IVR use. The change in relative or absolute abundance of vaginal bacterial phylotypes was similar among active and placebo IVR users (all q values >0.13). TFV concentrations in CV aspirate and vaginal tissue, and TFV-DP concentrations in vaginal tissue were not significantly different among users with community state type (CST) 4 versus those with Lactobacillus dominated microbiota (all p values >0.07). The proportions of participants with CV aspirate concentrations of TFV >200,000 ng/mL and those with tissue TFV-DP concentrations >1,000 fmol/mg were similar among women with anaerobe versus Lactobacillus dominated microbiota (p = 0.43, 0.95 respectively). There were no significant correlations between the CV aspirate concentration of TFV and the relative abundances of Gardnerella vaginalis or Prevotella species. Tissue concentrations of TFV-DP did not correlate with any the relative abundances of any species, including Gardnerella vaginalis. In conclusion, active IVRs did not differ from the placebo IVR on the effect on the vaginal microbiota. Local TFV and TFV-DP concentrations were high and similar among IVR users with Lactobacillus dominated microbiota versus CST IV vaginal microbiota. Trial registration: ClinicalTrials.gov NCT02235662.


Multiple Effects of Ascorbic Acid against Chronic Diseases: Updated Evidence from Preclinical and Clinical Studies.
Severe disease commonly manifests as a systemic inflammatory process. Inflammation is associated withthe enhanced production of reactive oxygen and nitrogen species and with a marked reduction in the plasma concentrations of protective antioxidant molecules. This imbalance gives rise to oxidative stress, which is greater in patients with more severe conditions such as sepsis, cancer, cardiovascular disease, acute respiratory distress syndrome, and burns. In these patients, oxidative stress can trigger cell, tissue, and organ damage, thus increasing morbidity and mortality. Ascorbic acid (ASC) is a key nutrient thatserves as an antioxidant and a cofactor for numerous enzymatic reactions. However, humans, unlike most mammals, are unable to synthesize it. Consequently, ASC must be obtained through dietary sources, especially fresh fruit and vegetables. The value of administering exogenous micronutrients, to reestablish antioxidant concentrations in patients with severe disease, has been recognized for decades. Despite the suggestion that ASC supplementation may reduce oxidative stress and prevent several chronic conditions, few large, randomized clinical trials have tested it in patients with severe illness. This article reviews the recent literature on the pharmacological profile of ASC and the role of its supplementation in critically ill patients.


Causal relationships among the gut microbiome, short-chain fatty acids and metabolic diseases.
Microbiome-wide association studies on large population cohorts have highlighted associations between the gut microbiome and complex traits, including type 2 diabetes (T2D) and obesity1. However, the causal relationships remain largely unresolved. We leveraged information from 952 normoglycemic individuals for whom genome-wide genotyping, gut metagenomic sequence and fecal short-chain fatty acid (SCFA) levels were available2, then combined this information with genome-wide-association summary statistics for 17 metabolic and anthropometric traits. Using bidirectional Mendelian randomization (MR) analyses to assess causality3, we found that the host-genetic-driven increase in gut production of the SCFA butyrate was associated with improved insulin response after an oral glucose-tolerance test (P = 9.8 × 10-5), whereas abnormalities in the production or absorption of another SCFA, propionate, were causally related to an increased risk of T2D (P = 0.004). These data provide evidence of a causal effect of the gut microbiome on metabolic traits and support the use of MR as a means to elucidate causal relationships from microbiome-wide association findings.


Current and frequency modulation characteristics for continuous-wave quantum cascade lasers at 9.06 μm.
We report the characteristics of current induced frequency modulation (FM) for two continuous-wave quantum cascade lasers (QCLs) at 9.06 μm. Both the frequency tuning rate and the phase shift between intensity modulation and FM are measured at different modulation frequencies from 10 Hz to 200 kHz. The frequency tuning rate of the QCLs depends on both the modulation frequency and amplitude. The tested QCL has been used to detect ambient water vapor with wavelength modulation spectroscopy for validation with a numerical model.


Ultrafast two-dimensional infrared vibrational echo chemical exchange experiments and theory.
Ultrafast two-dimensional (2D) infrared vibrational echo experiments and theory are used to examine chemical exchange between solute-solvent complexes and the free solute for the solute phenol and three solvent complex partners, p-xylene, benzene, and bromobenzene, in mixed solvents of the partner and CCl4. The experiments measure the time evolution of the 2D spectra of the hydroxyl (OD) stretching mode of the phenol. The time-dependent 2D spectra are analyzed using time-dependent diagrammatic perturbation theory with a model that includes the chemical exchange (formation and dissociation of the complexes), spectral diffusion of both the complex and the free phenol, orientational relaxation of the complexes and free phenol, and the vibrational lifetimes. The detailed calculations are able to reproduce the experimental results and demonstrate that a method employed previously that used a kinetic model for the volumes of the peaks is adequate to extract the exchange kinetics. The current analysis also yields the spectral diffusion (time evolution of the dynamic line widths) and shows that the spectral diffusion is significantly different for phenol complexes and free phenol.


Laugier-Hunziker syndrome in endocrine clinical practice.
Laugier-Hunziker syndrome (LHS) is a rare, benign and acquired disorder characterized by hyperpigmentation of the oral cavity and lips along with longitudinal melanonychia. No underlying systemic abnormalities or malignant predisposition is associated with it. In everyday clinical practice, an endocrinologist encounters certain endocrine conditions (e.g. Addison's disease, McCune-Albright syndrome) that present with, inter alia, mucocutaneous hyperpigmentation. Even though LHS is easily distinguished from endocrine entities mentioned earlier, diagnostic evaluation usually requires skilled and thorough practitioner. Since it is the diagnosis of exclusion, a number of systemic conditions must be ruled out prior to making the final diagnosis. However, its major differential diagnosis is primarily Peutz-Jeghers syndrome, which carries an increased risk of cancer. Here, we report a case of a young woman who was referred to the endocrinologist for diagnostic evaluation of dark-colored lesions of the oral cavity and nails. All performed laboratory tests were within reference range. Endoscopic gastrointestinal evaluation did not reveal neoplastic formations. Owing to an adult-onset, asymptomatic clinical course and negative diagnostic findings, we made a final diagnosis. In this case, target diagnostic evaluation notably reduced the need for additional expensive and invasive procedures and treatments. Laugier-Hunziker syndrome is a rare, acquired cause of asymptomatic, benign mucocutaneous hyperpigmentation.Prior to making a final diagnosis, certain medical entities with overlapping clinical features must be excluded.Endocrine conditions that usually present with the hyperpigmentation of the skin and mucous membranes (e.g. Addison's disease, McCune-Albright syndrome) can be easily ruled out based on clinical and laboratory findings.Its major differential diagnosis, Peutz-Jeghers syndrome is characterized by melanotic macules of the face and mouth, intestinal polyposis and significantly increased risk of different types of cancer, especially gastrointestinal.Anamnesis, physical examination and target diagnostic evaluation reduce the need for additional invasive and expensive procedures and treatment.


Relapse prevention in major depressive disorder: Mindfulness-based cognitive therapy versus an active control condition.
We evaluated the comparative effectiveness of mindfulness-based cognitive therapy (MBCT) versus an active control condition (ACC) for depression relapse prevention, depressive symptom reduction, and improvement in life satisfaction. Ninety-two participants in remission from major depressive disorder with residual depressive symptoms were randomized to either an 8-week MBCT or a validated ACC that is structurally equivalent to MBCT and controls for nonspecific effects (e.g., interaction with a facilitator, perceived social support, treatment outcome expectations). Both interventions were delivered according to their published manuals. Intention-to-treat analyses indicated no differences between MBCT and ACC in depression relapse rates or time to relapse over a 60-week follow-up. Both groups experienced significant and equal reductions in depressive symptoms and improvements in life satisfaction. A significant quadratic interaction (Group × Time) indicated that the pattern of depressive symptom reduction differed between groups. The ACC experienced immediate symptom reduction postintervention and then a gradual increase over the 60-week follow-up. The MBCT group experienced a gradual linear symptom reduction. The pattern for life satisfaction was identical but only marginally significant. MBCT did not differ from an ACC on rates of depression relapse, symptom reduction, or life satisfaction, suggesting that MBCT is no more effective for preventing depression relapse and reducing depressive symptoms than the active components of the ACC. Differences in trajectory of depressive symptom improvement suggest that the intervention-specific skills acquired may be associated with differential rates of therapeutic benefit. This study demonstrates the importance of comparing psychotherapeutic interventions to active control conditions.


Methods for objectively assessing clinical masticatory performance: protocol for a systematic review.
Chewing and masticatory function constitutes one of the most important oral health factors that affect quality of life, especially in older individuals. Little consensus currently exists regarding ways to objectively assess clinical masticatory performance (in this context, performance refers an individual's objective ability to mix or comminute food bolus). That said, many methods were developed to assess masticatory performance. Consequently, systematic review of the literature would be of great value when it comes to identifying various methods for objectively assessing clinical masticatory performance and for evaluating these methods. This study protocol describes a systematic review that intends to (i) identify methods to objectively assess clinical masticatory performance and (ii) evaluate psychometric properties (such as validity and reliability) of the identified methods. A systematic literature search is required to do so in these sources: MEDLINE (Ovid), Embase (embase.com), Web of Science Core Collection (Thomson Reuters), Cochrane (Wiley), and Cinahl (Ebsco). studies in scientific, full-text articles; development articles; validation articles; studies of the general adult population, ages ≥18. topics and article types that cover interview methods and self-reported questionnaires; methods/instruments that measure subjective masticatory performance; qualitative studies and case studies; opinion and editorial pieces; animal studies; studies of humans with severe oral health complications. This systematic review will result in a comprehensive assessment of various methods designed to objectively measure clinical masticatory performance. This systematic review will rate these methods, assess their reliability and validity, and identify one or more methods that can be recommended for use in clinical and scientific environments. From what is currently known, no systematic evaluation of various methods for objectively assessing clinical masticatory performance has been published. PROSPERO CRD42016037700.


Assessment of the prevalence of heart murmurs in overtly healthy cats.
To assess the frequency of heart murmurs in overtly healthy cats. Prospective study. 103 healthy domestic cats. Background information and physical characteristics were assessed in cats that were candidates for blood donation during an 8-month period. For cats with heart murmurs, additional information collected included murmur timing, grade, point of maximal intensity, and presence of additional heart sounds. Heart murmurs were detected in 22 of the 103 (21%) cats. Echocardiography was performed in 7 of those 22 cats. The echocardiogram was considered normal in 1 cat; in the other 6 cats, diagnoses included hypertrophic cardiomyopathy (interventricular septal hypertrophic form [IVSH]; n = 4), left ventricular concentric hypertrophy with valvular disease (1), and equivocal IVSH (1). Thirteen cats had more than 1 examination during the study; 3 of them developed heart murmurs. There were no significant differences in age, sex, breed, coat color, eye color, or heart rate between cats with and without murmurs. Among the 103 cats, there were 6 pairs of siblings from 6 multiple-cat households and 16 cats from 7 multiple-cat households in which the cats were not related; the proportion of cats with murmurs was higher in the related cats (5/12) than in the unrelated cats (3/16), but the difference was not significant. Results indicated that heart murmurs are detectable in a large proportion of overtly healthy cats and that many murmurs appear to be caused by structural heart disease that is in a clinically latent state.


Longitudinal MRI contrast enhanced monitoring of early tumour development with manganese chloride (MnCl2) and superparamagnetic iron oxide nanoparticles (SPIOs) in a CT1258 based in vivo model of prostate cancer.
Cell lines represent a key tool in cancer research allowing the generation of neoplasias which resemble initial tumours in in-vivo animal models. The characterisation of early tumour development is of major interest in order to evaluate the efficacy of therapeutic agents. Magnetic resonance imaging (MRI) based in-vivo characterisation allows visualisation and characterisation of tumour development in early stages prior to manual palpation. Contrast agents for MRI such as superparamagnetic iron oxide nanoparticles (SPIOs) and manganese chloride (MnCl2) represent powerful tools for the in-vivo characterisation of early stage tumours. In this experimental study, we labelled prostate cancer cells with MnCl2 or SPIOs in vitro and used 1 T MRI for tracing labelled cells in-vitro and 7 T MRI for tracking in an in-vivo animal model. Labelling of prostate cancer cells CT1258 was established in-vitro with MnCl2 and SPIOs. In-vitro detection of labelled cells in an agar phantom was carried out through 1 T MRI while in-vivo detection was performed using 7 T MRI after subcutaneous (s.c.) injection of labelled cells into NOD-Scid mice (n = 20). The animals were scanned in regular intervals until euthanization. The respective tumour volumes were analysed and corresponding tumour masses were subjected to histologic examination. MnCl2in-vitro labelling resulted in no significant metabolic effects on proliferation and cell vitality. In-vitro detection-limit accounted 105 cells for MnCl2 as well as for SPIOs labelling. In-vivo 7 T MRI scans allowed detection of 103 and 104 cells. In-vivo MnCl2 labelled cells were detectable from days 4-16 while SPIO labelling allowed detection until 4 days after s.c. injection. MnCl2 labelled cells were highly tumourigenic in NOD-Scid mice and the tumour volume development was characterised in a time dependent manner. The amount of injected cells correlated with tumour size development and disease progression. Histological analysis of the induced tumour masses demonstrated characteristic morphologies of prostate adenocarcinoma. To the best of our knowledge, this is the first study reporting direct in-vitro MnCl2 labelling and 7 T based in-vivo MRI tracing of cancer cells in a model of prostate cancer. MnCl2 labelling was found to be suitable for in-vivo tracing allowing long detection periods. The labelled cells kept their highly tumourigenic potential in-vivo. Tumour volume development was visualised prior to manual palpation allowing tumour characterisation in early stages of the disease.


Differentiating neurons activate transcription of the brain lipid-binding protein gene in radial glia through a novel regulatory element.
Formation and maintenance of a radial glial scaffold is fundamental for development of the vertebrate central nervous system. In mammals, radial glia arise in the neuroepithelium immediately prior to differentiation and migration of neurons away from the ventricular zones, and they are maintained until neuronal migration subsides. We have previously shown that expression of the brain lipid-binding protein (BLBP) in radial glia throughout the developing CNS is strictly correlated with the differentiation and migration of neurons upon these cells, and that BLBP function is required to maintain differentiation of primary cerebellar glial cells in vitro (Feng, L., Hatten, M. E. and Heintz, N. (1994). Neuron 12, 895-908). In this study, we demonstrate that BLBP transcription in vivo involves multiple regulatory elements, and that the dynamic temporal and spatial pattern of BLBP expression in radial and Bergmann glial cells throughout the developing CNS is programmed by a single radial glial cell-specific element (RGE). Furthermore, we demonstrate that BLBP expression in primary cerebellar glial cells requires coculture with differentiating neurons, and that this induction is regulated by the radial glia-specific element. The fact that transcription of BLBP in response to neurons in vitro and its dynamic regulation in radial glia throughout the CNS in vivo are both controlled by the RGE provides the first direct evidence supporting a role for differentiating neurons in the epigenetic regulation of radial glial cell function in vivo.


Exome sequencing identifies rare variants in multiple genes in atrioventricular septal defect.
The genetic etiology of atrioventricular septal defect (AVSD) is unknown in 40% cases. Conventional sequencing and arrays have identified the etiology in only a minority of nonsyndromic individuals with AVSD. Whole-exome sequencing was performed in 81 unrelated probands with AVSD to identify potentially causal variants in a comprehensive set of 112 genes with strong biological relevance to AVSD. A significant enrichment of rare and rare damaging variants was identified in the gene set, compared with controls (odds ratio (OR): 1.52; 95% confidence interval (CI): 1.35-1.71; P = 4.8 × 10(-11)). The enrichment was specific to AVSD probands, compared with a cohort without AVSD with tetralogy of Fallot (OR: 2.25; 95% CI: 1.84-2.76; P = 2.2 × 10(-16)). Six genes (NIPBL, CHD7, CEP152, BMPR1a, ZFPM2, and MDM4) were enriched for rare variants in AVSD compared with controls, including three syndrome-associated genes (NIPBL, CHD7, and CEP152). The findings were confirmed in a replication cohort of 81 AVSD probands. Mutations in genes with strong biological relevance to AVSD, including syndrome-associated genes, can contribute to AVSD, even in those with isolated heart disease. The identification of a gene set associated with AVSD will facilitate targeted genetic screening in this cohort.


Expression of delta-like ligand 4 (Dll4) and markers of hypoxia in colon cancer.
Delta-like ligand 4 (Dll4) is a Notch ligand that is upregulated by hypoxia and vascular endothelial growth factor-A (VEGF-A) and is reported to have a role in tumor angiogenesis. Evidence from xenograft studies suggests that inhibiting Dll4-Notch signalling may overcome resistance to anti-VEGF therapy. The aim of this study was to characterise the expression of Dll4 in colon cancer and to assess whether it is associated with markers of hypoxia and prognosis. In all, 177 colon cancers were represented in tissue microarrays. Immunohistochemistry was performed using validated antibodies against Dll4, VEGF, hypoxia-inducible factor (HIF)-1alpha, HIF-2alpha, prolyl hydroxylase (PHD)1, PHD2, PHD3 and carbonic anhydrase 9 (CA9). The expression of Dll4 was observed preferentially in the endothelium of 71% (125 out of 175) of colon cancers, but not in the endothelium adjacent to normal mucosa (none out of 107, P<0.0001). The expression of VEGF was significantly associated with HIF-2alpha (P<0.0001) and Dll4 (P=0.010). Only HIF-2alpha had a significant multivariate prognostic effect (hazard ratio 1.61, 95% confidence interval 1.01-2.57). Delta-like ligand 4 was also expressed by neoplastic cells, particularly neoplastic goblet cells. Endothelial expression of Dll4 is not a prognostic factor, but is significantly associated with VEGF. Assessing endothelial Dll4 expression may be critical in predicting response to anti-VEGF therapies.


Transcriptomic response of red grouse to gastro-intestinal nematode parasites and testosterone: implications for population dynamics.
A central issue in ecology is in understanding the relative influences of intrinsic and extrinsic effects on population regulation. Previous studies on the cyclic population dynamics of red grouse (Lagopus lagopus scoticus) have emphasized the destabilizing effects of either nematode parasites or territorial behaviour and aggression. The potential interacting effects of these processes, mediated through density-dependent, environmentally induced alterations of host immunocompetence influencing susceptibility to parasites have not been considered. Male red grouse at high density are more aggressive, associated with increased testosterone, which potentially could lead to reduced immunocompetence at a stage when parasites are most prevalent. This could depress individual condition, breeding performance and survival and thus drive or contribute to overall reductions in population size. Here, we characterize the transcriptomic response of grouse to nematode parasite infection and investigate how this is subsequently affected by testosterone, using a microarray approach contrasting red grouse with high and low parasite load at both high and low testosterone titre. A suite of 52 transcripts showed a significant level of up-regulation to either chronic parasite load or experimental parasite infection. Of these, 51 (98%) showed a reduced level of expression under conditions of high parasite load and high testosterone. The genes up-regulated by parasites and then down-regulated at high testosterone titre were not necessarily associated with immune response, as might be intuitively expected. The results are discussed in relation to the fitness and condition of individual red grouse and factors influencing the regulation of abundance in natural populations.


Efficacy of nicergoline in dementia and other age associated forms of cognitive impairment.
Nicergoline is an ergot derivative currently in use in over fifty countries for more than three decades, for the treatment of cognitive, affective, and behavioral disorders of older people. It was initially considered as a vasoactive drug and mainly prescribed for cerebrovascular disorders. Recent findings suggest other actions which has provided a rationale for the use of nicergoline for the treatment of various forms of dementia, including Alzheimer's Disease. To determine whether there is evidence of efficacy of nicergoline in the treatment of dementia and other age-associated forms of cognitive decline,and to assess the safety and tolerability of the drug. 1. Electronic databases search. The Cochrane Controlled Trials Register (which contains citations from the MEDLINE, EMBASE, Psych LIT, and hand searches of geriatric, dementia, psychogeriatric journals, and conference abstracts) was searched using the following terms: 'Nicergoline', 'Sermion'. 2. Reference search. The reference lists of all obtained studies was checked. 3. Pharmaceutical company Pharmacia & Upjohn, owners of the rights to produce and market nicergoline in various different countries, was asked to provide data and reports of clinical trials. In case of unavailability of numerical data in published studies, the authors of each paper, were asked for any published or unpublished data. - All unconfounded, double-blind, randomized, placebo-controlled, published and unpublished trials were sought. Non-randomized trials were excluded. Open trials were considered for inclusion if patients were randomized to the different treatment groups. - All patients diagnosed as having dementia or other cognitive disorder defined according to classification criteria accepted at the time of each study. - Nicergoline given at any dose for more than one day with placebo control. Type of outcome variables: 1. Cognitive function (as measured by psychometric tests). 2. Clinical impression (such as CIBIC or other clinical global measures of change). 3. Functional performance including dependency. 4. Behavioural disturbance. 5. Safety and acceptability as measured by the incidence of adverse effects (including side-effects) leading to withdrawal. 6. Death 7. Effect on carer 8. Use of services 9. Quality of life. A comprehensive search of the international literature and the producing company archives has been performed to identify all possible sources of data for this review. Only those trials fulfilling the inclusion criteria of belonging to either category A or B of allocation concealment, as defined by the Cochrane Organisation, were examined for data extraction by one reviewer. If there was doubt then the other reviewer was consulted. Data availability restricted analyses to 'completers' analyses for the outcome measures. Outcomes able to be assessed included: Behaviour, Cognition, Clinical Judgment, Tolerability, EEG. The Sandoz Clinical Assessment Geriatric Scale (SCAG) was the outcome used in the largest number of patients (814 patients). The results from these studies were homogeneous in nature despite including patients observed for periods of time ranging from 2 months to 12 months. There was a difference in favour of the active treatment in reducing the behavioural symptoms described by this scale, -5.18 points [-8.03, -2.33]. This scale has a maximum of 133 points. The therapeutic effects of nicergoline seem to be evident by 2 months of treatment and maintained for 6 months. In general other behavioural outcome measures which include the GRS, the IADL, and the MACC and were episodically used in few studies, failed to demonstrate statistically significant results although there was a trend favouring treatment. Cognitive assessment has been performed in a moderate number of patients with the MMSE (261 patients) and the ADAS-Cog (342 patients). No significant heterogeneity was found for these trials, despite the trials extending over periods of treatment of 3 to 12 months. There was a difference between treatment and control groups on the MMSE favouring nicergoline treatment. At 12 months the effect size was 2.86 [0.98, 4.74] The effect size for the ADAS-Cog, used exclusively with Alzheimer's disease patients, did not reveal a significant benefit. At 12 months the trend favoured treatment (-1.64 [-4.62, 1.34]). The other results from various cognitive measures tended to favour nicergoline but this was based on a small number of cases. The clinical impression of change obtained from a total of 921 patients was homogeneous across the studies, despite reflecting changes over periods of time ranging from 2 to 12 months. The Peto odd ratio for improvement in the subjects treated with nicergoline over these varying time periods was 3.33 [2.50, 4.43]. Tolerability assessed in 1427 patients was homogeneous across all studies and demonstrated a mildly increased risk of adverse events on treatment, OR 1.51[1.10, 2.07]. The clinical studies on nicergoline were carried out with diverse criteria and modalities of evaluation. Despite this, the 14 studies included in this review, have presented generally consistent results. Results of this meta-analysis provide some evidence of positive effects of nicergoline on cognition and behaviour and these effects are supported by an effect on clinical global impression. There was some evidence that there were increased risk of adverse effects associated with nicergoline. These results were obtained on older patients with mild to moderate cognitive and behavioural impairment of various clinical origins, including chronic cerebrovascular disorders and Alzheimer's dementia. The few studies specifically performed on patients with Alzheimer's disease were performed with too few people to give a definitive answer to the questions concerning the use of nicergoline for this form of dementia. This drug has not been evaluated using current diagnostic categories such as MCI or in association with therapeutic agents of different nature such as cholinesterase or antioxidant drugs.


Ameliorative effect of traditional Japanese medicine yokukansan on age-related impairments of working memory and reversal learning in rats.
Aging is thought to impair prefrontal cortical (PFC) structure-sensitive cognitive functions and flexibility, such as working memory and reversal learning. A traditional Japanese medicine, yokukansan (YKS), is frequently used to treat age-related neurodegenerative disorders such as Alzheimer's disease in Japan, but its pharmacological properties have not been elucidated. The present study was designed to examine whether YKS improves age-related cognitive deficits using aged rats. YKS was administered to 21-month-old rats for 3 months. The ability to learn initially a reward rule for a T-maze discrimination task (initial learning) was examined in young control (4-month-old), aged control (24-month-old) and YKS-treated aged (24-month-old) rats. Subsequently, working memory and reversal learning were examined in delayed alternation and reversal discrimination T-maze tasks, respectively. Locomotor activity was also measured in new environments. Although performance accuracy in the initial learning procedure did not differ among any experimental groups, accuracy in the delayed alternation task was significantly decreased in aged rats compared to young rats. Aged rats also showed significant decreases in accuracy in the reversal discrimination task. YKS treatment significantly ameliorated the age-related decreases in accuracy in the delayed alternation and reversal discrimination tasks. The ameliorative effects of YKS on impaired delayed alternation performance were reduced by intracranial infusions of a dopamine D1 receptor antagonist, SCH 23390, into the prelimbic cortical region of the PFC, and the YKS effects on impaired reversal learning were done by the infusions into the orbitofrontal cortex (OFC). Locomotor activity did not change in any experimental group. Thus, YKS ameliorated age-related impairments of working memory and reversal learning, which might be mediated by a dopaminergic mechanism in the PFC structure. These investigations provide information important for the treatment of brain dysfunctions in the elderly people.


c-Fos expression in the auditory pathways related to the significance of acoustic signals in rats performing a sensory-motor task.
Neuronal activity was established in the auditory pathways in relation to behavioural response and cognitive information processing during a sensory-motor acoustic learning. Rats were trained in three consecutive phases. The first phase was an association between an auditory stimulus and a food reward; the second phase a simple discrimination between two sounds of different frequency components, and the third phase a more complex discrimination involving both spectral and spatial sound dimensions. Auditory stimuli were bursts of complex sounds lasting 500 ms. Neuronal activity related to the behaviourally relevant stimuli was established in 20 "learning" rats undergoing this protocol, which were progressively sacrificed at the beginning, middle and end of each phase. For comparison, activity was also established in four "control" rats exposed to the same stimuli delivered pseudo-randomly, thus carrying no behavioural meaning. Neuronal activity was assessed immunocytochemically using the functional marker Fos. To establish a baseline, two rats were unexposed to controlled acoustic stimulation ("unstimulated" rats). In the superior olivary complex (SOC), inferior colliculus (IC) and medial geniculate body (MGB), the number of Fos-like immunopositive cells was comparable in "learning" and "control" animals, but higher than in the "unstimulated" rats. In the auditory cortex (AC), most prominently in the secondary area Te2, the number of Fos-like positive cells differed between "learning" and "control" rats, suggesting that the auditory cortical areas may be involved in the encoding of the behavioural significance of the acoustic stimuli.


Aging of cardiac myocytes in culture: oxidative stress, lipofuscin accumulation, and mitochondrial turnover.
Oxidative stress is believed to be an important contributor to aging, mainly affecting long-lived postmitotic cells such as cardiac myocytes and neurons. Aging cells accumulate functionally effete, often mutant and enlarged mitochondria, as well as an intralysosomal undegradable pigment, lipofuscin. To provide better insight into the role of oxidative stress, mitochondrial damage, and lipofuscinogenesis in postmitotic aging, we studied the relationship between these parameters in cultured neonatal rat cardiac myocytes. It was found that the content of lipofuscin, which varied drastically between cells, positively correlated with mitochondrial damage (evaluated by decreased innermembrane potential), as well as with the production of reactive oxygen species. These results suggest that both lipofuscin accumulation and mitochondrial damage have common underlying mechanisms, likely including imperfect autophagy and ensuing lysosomal degradation of oxidatively damaged mitochondria and other organelles. Increased size of mitochondria (possibly resulting from impaired fission due to oxidative damage to mitochondrial DNA, membranes, and proteins) also may interfere with mitochondrial turnover, leading to the appearance of so-called "giant" mitochondria. This assumption is based on our observation that pharmacological inhibition of autophagy with 3-methyladenine induced only moderate accumulation of large (senescent-like) mitochondria but drastically increased numbers of small, apparently normal mitochondria, reflecting their rapid turnover and suggesting that enlarged mitochondria are poorly autophagocytosed. Overall, our findings emphasize the importance of mitochondrial turnover in postmitotic aging and provide further support for the mitochondrial-lysosomal axis theory of aging.


Metabolic surgery in South Africa: an initial academic hospital experience.
In South Africa, 42.0% of adult females and 13.5% of adult males are classified as obese, the highest recorded numbers in Sub-Saharan Africa. Metabolic surgery has been proven to be a safe and effective treatment, yet due to demand on government resources has only been performed to a limited extent in public hospitals. The aim of this study was to describe the safety and efficacy of performing metabolic surgery at a single academic hospital in South Africa. This was a single centre retrospective review of 57 metabolic surgery procedures performed from October 2011 to September 2017 at Tygerberg Hospital, Cape Town, South Africa. The primary outcome was safety including mortality and adverse events. Secondary outcomes included effect of surgery on weight and diabetes resolution. A total of 57 patients underwent laparoscopic metabolic surgery, of which 44 (83.0%) were female with a mean age (standard deviation) of 42.8 (8.0) years. Fifty-six patients (98%) underwent Roux-and-Y gastric bypass and one (2%) had a sleeve gastrectomy performed. There were no mortalities and overall morbidity was 14.0%, with 3 (5.3%) classified as major and 5 (8.8%) as minor. The follow-up rate at 1 year was 100%. Mean preoperative body mass index (BMI) was 58.8 kg/m2, and comorbidities included hypertension (59.6%), Type 2 Diabetes (42.1%), and dyslipidaemia (36.8%). There were no conversions to open surgery and at one year the mean (95% confidence interval) percentage excess body mass index loss was 50.4% (44.0-56.8%). Metabolic surgery can be performed safely in the public sector in South Africa, with short-term safety and efficacy outcomes comparable to international reports. Larger scale studies are needed to determine long-term outcomes and cost-effectiveness.


Temporary Transfer of Firearms From the Home to Prevent Suicide: Legal Obstacles and Recommendations.
The presence of firearms in the home increases the risk of suicide for residents. As a result, clinicians and professional organizations recommend counseling about temporary removal of firearms from the home of potentially suicidal individuals. In some states, however, firearm laws may affect the ability to easily transfer a gun temporarily to reduce suicide risk. In particular, universal background check (UBC) laws-which require a background check whenever a gun is transferred, even by non-gun dealers-may also apply to temporary transfers intended to reduce suicide risk. Clinicians have previously reported that confusion regarding state firearm laws and uncertainty over the legality of a temporary transfer have affected their ability to effectively counsel patients. We summarize the laws of all 50 states and specifically examine the relevant firearm laws of 3 representative states with UBCs and different approaches-Maryland, Colorado, and California. We identify both helpful and problematic aspects of state laws regarding temporary transfer of firearms. We provide recommendations for amending UBC laws to make it easier for clinicians and patients to temporarily transfer firearms.


Educational inequalities in smoking cessation trends in Italy, 1982-2002.
Smoking prevalence rates are declining in most industrialised countries, partly because of growing cessation rates. However, little is known on recent time-trends in smoking cessation by socioeconomic position. This study aims to estimate educational inequalities in smoking cessation trends in Italy between 1982 and 2002. Data were derived from two national health interview surveys carried out in Italy in 1999-2000 (n = 34 789) and in 2004-2005 (n = 33 135). On the basis of respondents' age at starting and age at quitting smoking, we computed age-standardised smoking cessation rates at ages 20-44 years for subjects who were current smokers between 1982 and 2002. Smoking quit rates were approximately constant at a figure of about 2 per 100 person-years until the period 2000-2002, when they rapidly increased up to 3-4 per 100 person-years. Higher educated smokers constantly showed higher cessation rates than lower educated subjects (rate ratio 1.33; 95% CI 1.25 to 1.41 for men and 1.41; 95% CI 1.30 to 1.53 for women). The relative size of educational difference in smoking cessation did not significantly vary by period. However, in absolute terms, the increase in cessation rates in 2000-2002 was larger among higher educated smokers. Educational inequalities in smoking cessation persisted in both relative and absolute terms. The increase in smoking cessation rates in 2000-2002 suggests that tobacco control policies may have reached more disadvantaged smokers, although smokers of higher socioeconomic groups seem to have benefited the most.


Pigment Epithelial-Derived Factor Peptide Facilitates the Regeneration of a Functional Limbus in Rabbit Partial Limbal Deficiency.
To investigate the potential of a pigment epithelial-derived factor (PEDF) peptide 44-mer to promote limbal regeneration in a rabbit partial limbal deficiency model. Limbal excision (180°) was created surgically, and topical application of 44-mer-containing ointment once a day for 2 weeks was started immediately after injury. Limbal barrier function was inspected at 2 and 6 months after treatment. Corneal neovascularization was observed under slit-lamp microscope. The presence of goblet cells on the corneal surface was examined using impression cytology. The resulting repair tissue was assessed by immunohistochemical staining with antibodies for putative limbal stem cell (LSC) markers ΔNp63α and ABCG2. Cells harvested from the regenerated tissue were analyzed for colony-forming capacity and expression of LSC markers by immunostaining assay and quantitative real-time PCR (qPCR). Eyes treated with the 44-mer blocked vascularization and goblet cell migration onto the corneal surface. By means of immunohistochemical staining and cell isolation in the repair tissue, we showed that LSCs were widely distributed at the regenerated tissue after 44-mer treatment. The repaired limbus contributed robustly to corneal wound healing as effectively as undamaged limbus. We demonstrated that 44-mer regenerates a functional limbus-like structure on limbal excision wounds. Our finding suggests that the PEDF peptide derivative may be an innovative strategy for tissue engineering and repair therapy in partial LSC deficiency diseases.


Medial basal hypothalamic monoamine activity associated with intracerebroventricular p-chlorophenylalanine-induced hyperphagia.
There is evidence for reciprocal interactions between the brain monoamine neurotransmitters serotonin and noradrenaline which may play a critical role in homeostasis. The aim of the present study was to establish the effect of drug-induced damage to the serotoninergic system on noradrenergic activity in the hypothalamus. Bilateral intracerebroventricular injections of p-chlorophenylalanine (PCPA; 3 mg/kg in 2 x 6 microliters) were made to induce destruction in the serotoninergic system. Relative to saline-injected controls, PCPA-injected rats began overeating by 3 days postinjection. On day 10, when the experimental rats were consuming approximately 120% that of controls, animals were 4-h food deprived, sacrificed and the medial basal hypothalamus was removed for later analysis (by gas chromatography/mass spectrometry) of noradrenaline (NA), serotonin (5-HT) and dopamine (DA) and their principal metabolites dihydroxyphenylethyleneglycol (DHPG), 5-hydroxyindoleacetic acid (5-HIAA) and 3,4-dihydroxyphenylacetic acid (DOPAC), respectively. The ratio of metabolite to monoamine provided an index of functional activity. Trunk blood was collected for analysis of serum insulin and glucose. PCPA-injected animals had higher levels of DHPG (P less than 0.05), an increase in the DHPG/NA ratio (P less than 0.02), lower serum insulin (P less than 0.05) and increased serum glucose (P less than 0.05). There were significant correlations between noradrenergic activity (DHPG/NA ratio) and: (1) food intake (day 9 and 10 average; r = 0.62, P less than 0.05); and (2) serum glucose (r = 0.59, P less than 0.05).(ABSTRACT TRUNCATED AT 250 WORDS)


Phospholipid monolayers probed by vibrational sum frequency spectroscopy: instability of unsaturated phospholipids.
The surface specific technique vibrational sum frequency spectroscopy has been applied to in situ studies of the degradation of Langmuir monolayers of 1,2-diacyl-phosphocholines with various degrees of unsaturation in the aliphatic chains. To monitor the degradation of the phospholipids, the time-dependent change of the monolayer area at constant surface pressure and the sum frequency intensity of the vinyl CH stretch at the carbon-carbon double bonds were measured. The data show a rapid degradation of monolayers of phospholipids carrying unsaturated aliphatic chains compared to the stable lipids carrying fully saturated chains when exposed to the ambient laboratory air. In addition, the degradation of the phospholipids can be inhibited by purging the ambient air with nitrogen. This instability may be attributed to spontaneous degradation by oxidation mediated by various reactive species in the air. To further elucidate the process of lipid oxidation in biological membranes artificial Langmuir monolayers probed by a surface specific spectroscopic technique as in this study can serve as a model system for studying the degradation/oxidation of cell membrane constituents.


Doubling of the known set of RNA viruses by metagenomic analysis of an aquatic virome.
RNA viruses in aquatic environments remain poorly studied. Here, we analysed the RNA virome from approximately 10 l water from Yangshan Deep-Water Harbour near the Yangtze River estuary in China and identified more than 4,500 distinct RNA viruses, doubling the previously known set of viruses. Phylogenomic analysis identified several major lineages, roughly, at the taxonomic ranks of class, order and family. The 719-member-strong Yangshan virus assemblage is the sister clade to the expansive class Alsuviricetes and consists of viruses with simple genomes that typically encode only RNA-dependent RNA polymerase (RdRP), capping enzyme and capsid protein. Several clades within the Yangshan assemblage independently evolved domain permutation in the RdRP. Another previously unknown clade shares ancestry with Potyviridae, the largest known plant virus family. The 'Aquatic picorna-like viruses/Marnaviridae' clade was greatly expanded, with more than 800 added viruses. Several RdRP-linked protein domains not previously detected in any RNA viruses were identified, such as the small ubiquitin-like modifier (SUMO) domain, phospholipase A2 and PrsW-family protease domain. Multiple viruses utilize alternative genetic codes implying protist (especially ciliate) hosts. The results reveal a vast RNA virome that includes many previously unknown groups. However, phylogenetic analysis of the RdRPs supports the previously established five-branch structure of the RNA virus evolutionary tree, with no additional phyla.


Enterocolic lymphocytic phlebitis of the cecal pole and appendix vermiformis with increase of IgG4-positive plasma cells.
Here we describe the clinicopathological course of a 20-year-old female patient with enterocolic lymphocytic phlebitis (ELP) of the appendix vermiformis and cecal pole with increase of IgG4-positive plasma cells. The patient presented with acute abdomen, suspicious of acute appendicitis. Diagnostic laparoscopy showed tumefaction of the cecal pole and appendix vermiformis. Histologic examination revealed mural thickening and a dense lymphoplasmocytic, partly obliterative infiltrate of the veins with sparing of the arteries, diagnostic of ELP. In addition, we found an elevated number of IgG4-positive plasma cells blended in with the lymphocytes. The IgG4-to-IgG ratio accounted for >40 %. This case meets the histopathological criteria requested for IgG4-related disease (IgG4-RD) and thus opens the possibility that ELP might be part of the IgG4-RD spectrum.


Methodological quality of economic evaluations of new pharmaceuticals in The Netherlands.
In the Netherlands, decisions about the reimbursement of new pharmaceuticals are based on cost effectiveness, as well as therapeutic value and budget impact. Since 1 January 2005, drug manufacturers are formally required to substantiate the cost effectiveness of drugs that have therapeutic added value in comparison with existing ones through pharmacoeconomic evaluations. Dutch guidelines for pharmacoeconomic research provide methods guidance, ensuring consistency in both the evidence and the decision-making process about drug reimbursement. This study reviewed the methodological quality of all 21 formally required pharmacoeconomic evaluations of new pharmaceuticals between 1 January 2005 and 1 October 2008, and verified whether these evaluations complied with pharmacoeconomic guidelines. Data on the quality of the pharmacoeconomic evaluations were extracted from the pharmacoeconomic reports published by the Dutch Health Care Insurance Board (CVZ). The Board's newsletters provided information on the advice to, and reimbursement decisions made by, the Dutch Minister of Health. All data extraction was carried out by two independent reviewers, and descriptive analyses were conducted. The methodological quality was sound in only 8 of the 21 pharmacoeconomic evaluations. In most cases, the perspective of analysis, the comparator drugs, and the reporting of both total and incremental costs and effects were correct. However, drug indication, form (i.e. cost utility/cost effectiveness) and time horizon of the evaluations were frequently flawed. Moreover, the costs and effects of the pharmaceuticals were not always analysed correctly, and modelling studies were often non-transparent. Twelve drugs were reimbursed, and nine were not. The compliance with pharmacoeconomic guidelines in economic evaluations of new pharmaceuticals can be improved. This would improve the methodological quality of the pharmacoeconomic evaluations and ensure consistency in the evidence and the decision-making process for drug reimbursement in the Netherlands.


Benefits to faculty involved in medical school learning communities.
Job satisfaction plays a large role in enhancing retention and minimizing loss of physicians from careers in academic medicine. The authors explored the effect of learning communities (LCs) on the faculty members' job satisfaction. Between October 2011 and May 2012, the authors surveyed 150 academic clinical faculty members serving as LC mentors for students at five US medical schools. Factor analysis was used to explore satisfaction themes and relationships between these themes and other characteristics. Factor analysis revealed two major sources of this satisfaction: a Campus Engagement factor (e.g., feeling happier, improved sense of community, better communication skills, and feeling more productive) and a skills factor (e.g., improved clinical skills, being a better doctor). Higher Campus Engagement factor satisfaction was associated with less desire to leave the learning community (p = 0.01) and more FTE support for role in LC (p = 0.01). Higher skills factor satisfaction was associated with the school that provided more structured faculty development (p = 0.0001). Academic clinical faculty members reported serving as a mentor in an LC was a strong source of job satisfaction. LC may be a tool for retaining clinical faculty members in academic careers.


Clinical and CT features in pediatric patients with COVID-19 infection: Different points from adults.
To discuss the different characteristics of clinical, laboratory, and chest computed tomography (CT) in pediatric patients from adults with 2019 novel coronavirus (COVID-19) infection. The clinical, laboratory, and chest CT features of 20 pediatric inpatients with COVID-19 infection confirmed by pharyngeal swab COVID-19 nucleic acid test were retrospectively analyzed during 23 January and 8 February 2020. The clinical and laboratory information was obtained from inpatient records. All the patients were undergone chest CT in our hospital. Thirteen pediatric patients (13/20, 65%) had an identified history of close contact with COVID-19 diagnosed family members. Fever (12/20, 60%) and cough (13/20, 65%) were the most common symptoms. For laboratory findings, procalcitonin elevation (16/20, 80%) should be pay attention to, which is not common in adults. Coinfection (8/20, 40%) is common in pediatric patients. A total of 6 patients presented with unilateral pulmonary lesions (6/20, 30%), 10 with bilateral pulmonary lesions (10/20, 50%), and 4 cases showed no abnormality on chest CT (4/20, 20%). Consolidation with surrounding halo sign was observed in 10 patients (10/20, 50%), ground-glass opacities were observed in 12 patients (12/20, 60%), fine mesh shadow was observed in 4 patients (4/20, 20%), and tiny nodules were observed in 3 patients (3/20, 15%). Procalcitonin elevation and consolidation with surrounding halo signs were common in pediatric patients which were different from adults. It is suggested that underlying coinfection may be more common in pediatrics, and the consolidation with surrounding halo sign which is considered as a typical sign in pediatric patients.


[Childhood bacterial meningitis trends in Japan from 2009 to 2010].
We conducted a pediatric survey of bacterial meningitis epidemiology from January 2009 to December 2010 in Japan, and obtained the following results for 314 cases (186 boys, 124 girls, and 4 with gender not reported). Children younger than one year old accounted for the majority of cases (51.2%, 161/314), and the incidence decreased with increasing age. Haemophilus influenzae (in children aged 1 month to 5 years old) was the most common cause of infection (53.2%), followed by Streptococcus pneumoniae (1 month to 12 years, 24.2%), Streptococcus agalactiae (0-4 months, 7.6%), and Escherichia coli (0-3 months, 3.2%). Susceptibility tests showed that 50.1% (78/153) of the H. influenzae isolates and 63.0% (46/73) of the S. pneumoniae isolates were drug-resistant. Combinations of ampicillin and cephem or carbapenem and other beta-lactams were mainly used as the initial antibiotics for patients under 4 months of age (77.8%, 42/54), and a carbapenem and other beta-lactam combination was used for patients aged 4 months and older (76.4%, 198/259). The final antibiotics for H. influenzae and S. pneumoniae were mainly cefotaxime (CTX) or ceftriaxone (CTRX) and carbapenem, respectively. The overall fatality rate was 2.0% (6/305). Since the Haemophilus influenzae type b vaccine (Hib vaccine) and the 7 valent pneumococcal conjugate vaccine (PCV7) are not widely used in Japan, only 5 patients in our cohort (all with meningitis not caused by H. influenzae) had been immunized with the Hib vaccine, and none had been immunized with the PCV7 vaccine. No remarkable changes in the characteristics of pediatric meningitis have been observed for several years in Japan.


Deletion of Vhlh in chondrocytes reduces cell proliferation and increases matrix deposition during growth plate development.
The von Hippel Lindau tumor suppressor protein (pVHL) is a component of a ubiquitin ligase that promotes proteolysis of the transcription factor hypoxia-inducible-factor 1alpha (HIF1alpha), the key molecule in the hypoxic response. We have used conditional inactivation of murine VHL (Vhlh) in all cartilaginous elements to investigate its role in endochondral bone development. Mice lacking Vhlh in cartilage are viable, but grow slower than control littermates and develop a severe dwarfism. Morphologically, Vhlh null growth plates display a significantly reduced chondrocyte proliferation rate, increased extracellular matrix, and presence of atypical large cells within the resting zone. Furthermore, stabilization of the transcription factor HIF1alpha leads to increased expression levels of HIF1alpha target genes in Vhlh null growth plates. Lastly, newborns lacking both Vhlh and Hif1a genes in growth plate chondrocytes display essentially the same phenotype as Hif1a null single mutant mice suggesting that the Vhlh null phenotype could result, at least in part, from increased activity of accumulated HIF1alpha. This is the first study reporting the novel and intriguing findings that pVHL has a crucial role in endochondral bone development and is necessary for normal chondrocyte proliferation in vivo.


Perinatal risk factors for hospitalization for pneumococcal disease in childhood: a population-based cohort study.
The objective of this study was to examine the relation of factors that are present at birth to subsequent hospitalization for childhood pneumococcal disease. We conducted a cohort study of all singletons born in 3 counties in western Denmark from 1980 through 2001, using population-based registries to obtain data on pregnancy- and birth-related variables and hospitalizations through age 12. We calculated incidence rates of pneumococcal disease hospitalization overall and within strata of study variables and used Poisson regression to estimate rate ratios for pneumococcal disease hospitalization while accounting for other birth characteristics. Among 338,504 eligible births, 1052 children were later hospitalized for pneumococcal disease. Pneumonia accounted for most hospitalizations (81.9%). The pneumococcal disease hospitalization rate was highest among 7- to 24-month-olds, followed by 0- to 6-month-olds and 25- to 60-month-olds. The highest rates, typically over 200 hospitalizations per 100,000 person-years, were in 0- to 6- and 7- to 24-month-old children who were born preterm or with low birth weight, a low 5-minute Apgar score, or birth defects. The hospitalization rate was lower for first-born children at 0 to 6 months but not at older ages. At older ages, hospitalization rates were not substantially different for children whose mothers smoked during pregnancy, but at 0 to 6 months, the rate was higher for children of multiparous nonsmokers than for others. Adjusted rate ratios were elevated across all age categories for several variables, including low birth weight, presence of birth defects, and low 5-minute Apgar. For several others, including preterm birth, maternal multiparity, age < or = 20 years, and non-Danish/European Union citizenship, adjusted rate ratios were elevated only for 0- to 6-month-olds. This large cohort study of hospitalization for childhood pneumococcal disease clarifies the roles of some gestation and birth factors while raising new questions about how these factors work.


In vivo EPR on spin labeled colicin A reveals an oligomeric assembly of the pore-forming domain in E. coli membranes.
We report on the application of site-directed spin labeling (SDSL) and electron paramagnetic resonance (EPR) spectroscopy to study possible oligomerization of the bacterial toxin colicin A (ColA) upon membrane insertion in vitro and in vivo. We applied SDSL-EPR protocols and optimized experimental conditions to perform continuous wave EPR experiments and double electron-electron resonance distance measurements on intact Escherichia coli cells interacting with nitroxide spin-labeled ColA. Our data suggest that ColA forms dimers upon membrane insertion, thus explaining previously reported pore diameters of about 1 nm, which are unlikely to be formed by a single colicin A monomer.


Ant navigation: resetting the path integrator.
Desert ants use path integration as their predominant system of long-distance navigation, but they also make use of route-defining and nest-defining visual landmarks. Such landmark-gained information might override the information provided by the path integrator, but nevertheless the path integrator keeps running. Here we show that only cues that are associated with the ant being inside the nest are able to reset the path integrator to zero state. Ants were captured at a feeder, i.e. without having run off their home vector, and were forced to enter the nest. On their next outbound run their walking direction differed by 180 degrees from that of ants that had also been captured at the feeder but instead of having been forced to enter the interior of the nest were released at its entrance. Whereas these latter ants still ran off their home vector pointing in the feeder-nest direction (and by this departed from the nest in a direction opposite to the feeder direction) the former ants had reset their home vector to zero state, and had therefore been able to reload their learned feeder vector, and consequently departed from the nest in the feeder direction. Owing to its egocentric nature the path-integrator is error prone. Hence, it is a suitable strategy to reset the path integrator if the ant has appeared at its final goal, the nest. Otherwise during consecutive foraging runs navigational errors would steadily increase.


Beliefs about personal control and self-management in 30-40 year olds living with Inflammatory Bowel Disease: a qualitative study.
Inflammatory Bowel Disease is a collective term for two distinct long term conditions: Ulcerative Colitis and Crohn's disease. There is increasing emphasis on patients taking greater personal control and self-management of this condition, reflecting earlier research into the management of chronic illness. Nurses play a pivotal role in this process, yet how optimal personal control is self-assessed and self-managed in Inflammatory Bowel Disease is poorly understood. This study set out to explore beliefs about personal control and self-management of Inflammatory Bowel Disease. It focused on the role of physical, psychological and socio-economic factors within the individual's life experience. A qualitative approach was used comprising 24, one-to-one, semi-structured interviews with participants aged 30-40 years. Participants with a histological diagnosis of Inflammatory Bowel Disease for at least 12 months were eligible and recruited by gastrointestinal specialist staff from outpatient clinics at a large National Health Service Trust in the United Kingdom. Interviews were transcribed verbatim. Data analysis was informed by existing theories of personal control and used the 'systematic framework analysis' approach. In addition to existing theories of personal control, self-discrepancy theory helped to explain how people viewed the control and self-management of Inflammatory Bowel Disease. One main theme emerged from the findings: 'Reconciliation of the self in IBD', this was supported by three sub-themes and eight basic themes. Some participants found that being unable to control and predict the course of their condition was distressing, however for others this limited control was not viewed as a negative outcome. Being able to share control of IBD with specialist health care staff was beneficial, and participants stated that other priorities in life were as equally important to manage and control. A key barrier to ensuring greater personal control and self-management was a lack of knowledge and awareness by non-specialist health care staff, employers and the wider society. Nurses involved in the care of individuals with Inflammatory Bowel Disease should support and prepare patients for the discrepancies and uncertainties of living with the condition. Greater training about Inflammatory Bowel Disease is recommended, specifically for non-specialist health care staff and employers.


Extent of mangrove nursery habitats determines the geographic distribution of a coral reef fish in a South-Pacific archipelago.
Understanding the drivers of species' geographic distribution has fundamental implications for the management of biodiversity. For coral reef fishes, mangroves have long been recognized as important nursery habitats sustaining biodiversity in the Western Atlantic but there is still debate about their role in the Indo-Pacific. Here, we combined LA-ICP-MS otolith microchemistry, underwater visual censuses (UVC) and mangrove cartography to estimate the importance of mangroves for the Indo-Pacific coral reef fish Lutjanus fulviflamma in the archipelago of New Caledonia. Otolith elemental compositions allowed high discrimination of mangroves and reefs with 83.8% and 98.7% correct classification, respectively. Reefs were characterized by higher concentrations of Rb and Sr and mangroves by higher concentrations of Ba, Cr, Mn and Sn. All adult L. fulviflamma collected on reefs presented a mangrove signature during their juvenile stage with 85% inhabiting mangrove for their entire juvenile life (about 1 year). The analysis of 2942 UVC revealed that the species was absent from isolated islands of the New Caledonian archipelago where mangroves were absent. Furthermore, strong positive correlations existed between the abundance of L. fulviflamma and the area of mangrove (r = 0.84 for occurrence, 0.93 for density and 0.89 for biomass). These results indicate that mangrove forest is an obligatory juvenile habitat for L. fulviflamma in New Caledonia and emphasize the potential importance of mangroves for Indo-Pacific coral reef fishes.


YphC and YsxC GTPases assist the maturation of the central protuberance, GTPase associated region and functional core of the 50S ribosomal subunit.
YphC and YsxC are GTPases in Bacillus subtilis that facilitate the assembly of the 50S ribosomal subunit, however their roles in this process are still uncharacterized. To explore their function, we used strains in which the only copy of the yphC or ysxC genes were under the control of an inducible promoter. Under depletion conditions, they accumulated incomplete ribosomal subunits that we named 45SYphC and 44.5SYsxC particles. Quantitative mass spectrometry analysis and the 5-6 Å resolution cryo-EM maps of the 45SYphC and 44.5SYsxC particles revealed that the two GTPases participate in the maturation of the central protuberance, GTPase associated region and key RNA helices in the A, P and E functional sites of the 50S subunit. We observed that YphC and YsxC bind specifically to the two immature particles, suggesting that they represent either on-pathway intermediates or that their structure has not significantly diverged from that of the actual substrate. These results describe the nature of these immature particles, a widely used tool to study the assembly process of the ribosome. They also provide the first insights into the function of YphC and YsxC in 50S subunit assembly and are consistent with this process occurring through multiple parallel pathways, as it has been described for the 30S subunit.


Association of alcohol consumption with the risk of stomach cancer in a Japanese population: a prospective cohort study.
Biological studies have provided confirmation of alcohol-related carcinogenesis in the stomach, but the association between alcohol consumption and the risk of stomach cancer remains controversial. We aimed to investigate whether quantitative alcohol intake is associated with the risk of stomach cancer in a large prospective cohort study among a Japanese population. Study participants included 30 714 participants (14 171 men and 16 543 women) aged 35 years or older, who were enrolled in the Takayama study launched on 1 September 1992. Alcohol consumption was assessed quantitatively using a validated food frequency questionnaire. According to alcohol intake (g/day), male participants were classified into quartile groups: Q1, Q2, Q3, or Q4. Female participants were classified into three groups: nondrinkers, and drinkers below or above the median alcohol level. We estimated the hazard ratios and 95% confidence intervals (CIs) for stomach cancer adjusted for age, smoking, BMI, education, total energy intake, salt intake, physical activity, and medical history of diabetes mellitus for each alcohol intake group using the Cox proportional hazards regression model. By the end of March 2008, a total of 678 participants had been diagnosed with stomach cancer. For men, the multivariate-adjusted HRs of stomach cancer for Q2, Q3, and Q4 relative to Q1 were 1.39 (95% CI: 1.07-1.81), 1.35 (95% CI: 1.02-1.79), and 1.38 (95% CI: 1.02-1.87), respectively. In women, no associations were observed. These data suggest that alcohol consumption could be associated with an increased risk of stomach cancer among Japanese men.


GERD evaluation: time for a new paradigm?
In the past there has been reliance on heartburn and acid regurgitation as the cardinal symptoms of gastro-esophageal reflux. Now it is recognized that diverse additional symptoms including extraesophageal manifestations are essential components of gastroesophageal reflux disease (GERD). Furthermore, previous reliance in mucosal damage at endoscopy as evidence of therapeutic success in the management of GERD has proven of only limited value in the diverse world of GERD symptoms. Indeed is now apparent that the relationship between symptoms and lesions is modest at best. Most patients with GERD have no overt erosions or ulceration at endoscopy and they frequently suffer from a wide array of complaints other than heartburn that contribute to their diminished perception of life's quality. To address these problems, multidimensional questionnaires designed to encompass the diverse symptomatology of GERD have been investigated, principally, to provide a reproducible metric of change in the subjective response to pharmacologic therapy during trials for patients with nonerosive reflux disease. The most promising of these instruments is The ReQuest questionnaire, because it has been extensively validated in multiple patient groups and cultures, appears highly promising in this regard and for erosive disease too and may ultimately prove to be of value for the initial evaluation of patients with GERD. We propose that accurate symptom delineation complemented with endoscopy performed in selected groups of patients will become the mainstay in diagnosis of GERD and the assessment of therapy efficacy.


Rupture of granular rafts: effects of particle mobility and polydispersity.
Reversible encapsulation of liquid materials is a technical challenge in many applications such as for the transport and controlled delivery of active ingredients. In contrast to most state-of-the-art processes, capillary adsorbed solid particles can achieve chemical-reaction-free encapsulation by forming dense rafts which isolate the liquid from its surroundings. While the production conditions of such capsules have been characterized, the control of the armor robustness remains poorly described and understood. In this paper, we probe the armor robustness via impacts of droplets on encapsulated materials. Thereby, we establish the mechanisms and conditions of armor rupture and derive models that predict the rupturing thresholds or probabilities. Using monodisperse sized particles and gradually increasing the impacting drop velocity, a sharp transition from sustained to coalescing drops is observed. On mobile rafts made of particles at the water/air interface, the velocity threshold increases with increasing particle diameter while an opposite trend is observed on immobile rafts made of particles trapped at a gelified interface. Two models based on particle pair and triplet interactions, respectively, quantitatively match the experiments. Assembling rafts with particles of two different sizes significantly smoothens the coalescence transition, regardless of particle mobility. Beyond apparent similarities, rationalizing the rupturing probability of mobile and immobile armor evidences very different sensitivity to heterogeneities. On immobile armor, drop coalescence remains random and thus well described by the statistical particle distribution while on mobile armor the ruptures are preferably localized at the non-percolated parts of the granular network.


Neutrophils Promote Mycobacterial Trehalose Dimycolate-Induced Lung Inflammation via the Mincle Pathway.
Trehalose 6,6'-dimycolate (TDM), a cord factor of Mycobacterium tuberculosis (Mtb), is an important regulator of immune responses during Mtb infections. Macrophages recognize TDM through the Mincle receptor and initiate TDM-induced inflammatory responses, leading to lung granuloma formation. Although various immune cells are recruited to lung granulomas, the roles of other immune cells, especially during the initial process of TDM-induced inflammation, are not clear. In this study, Mincle signaling on neutrophils played an important role in TDM-induced lung inflammation by promoting adhesion and innate immune responses. Neutrophils were recruited during the early stage of lung inflammation following TDM-induced granuloma formation. Mincle expression on neutrophils was required for infiltration of TDM-challenged sites in a granuloma model induced by TDM-coated-beads. TDM-induced Mincle signaling on neutrophils increased cell adherence by enhancing F-actin polymerization and CD11b/CD18 surface expression. The TDM-induced effects were dependent on Src, Syk, and MAPK/ERK kinases (MEK). Moreover, coactivation of the Mincle and TLR2 pathways by TDM and Pam3CSK4 treatment synergistically induced CD11b/CD18 surface expression, reactive oxygen species, and TNFα production by neutrophils. These synergistically-enhanced immune responses correlated with the degree of Mincle expression on neutrophil surfaces. The physiological relevance of the Mincle-mediated anti-TDM immune response was confirmed by defective immune responses in Mincle⁻/⁻ mice upon aerosol infections with Mtb. Mincle-mutant mice had higher inflammation levels and mycobacterial loads than WT mice. Neutrophil depletion with anti-Ly6G antibody caused a reduction in IL-6 and monocyte chemotactic protein-1 expression upon TDM treatment, and reduced levels of immune cell recruitment during the initial stage of infection. These findings suggest a new role of Mincle signaling on neutrophils during anti-mycobacterial responses.


Visual detection of DNA on paper chips.
On-site DNA analysis for diagnostic or forensic purposes is much anticipated in the future of molecular testing. Yet the challenges to achieve this goal remain large with rapid and inexpensive detection and visualization being key factors for any portable analysis system. We have developed a filter paper-based nucleic acid assay, which is able to identify and distinguish dog and human genomic and mitochondrial samples in a forensic setting. The filter paper material allows for transport by capillary force of the sample DNA through the detection surface, allowing the targets to hybridize specifically to their complementary capture sequences. Coupling micrometer-sized beads to DNA allows the results to be visualized by the naked eye, enabling instant, cost-efficient, and on-site detection, while eliminating the need for advanced expensive instrumentation.


Design and development of an injection-molded demultiplexer for optical communication systems in the visible range.
Optical simulation software based on the ray-tracing method offers easy and fast results in imaging optics. This method can also be applied in other fields of light propagation. For short distance communications, polymer optical fibers (POFs) are gradually gaining importance. This kind of fiber offers a larger core diameter, e.g., the step index POF features a core diameter of 980 μm. Consequently, POFs have a large number of modes (>3 million modes) in the visible range, and ray tracing could be used to simulate the propagation of light. This simulation method is applicable not only for the fiber itself but also for the key components of a complete POF network, e.g., couplers or other key elements of the transmission line. In this paper a demultiplexer designed and developed by means of ray tracing is presented. Compared to the classical optical design, requirements for optimal design differ particularly with regard to minimizing the insertion loss (IL). The basis of the presented key element is a WDM device using a Rowland spectrometer setup. In this approach the input fiber carries multiple wavelengths, which will be divided into multiple output fibers that transmit only one wavelength. To adapt the basic setup to POF, the guidance of light in this element has to be changed fundamentally. Here, a monolithic approach is presented with a blazed grating using an aspheric mirror to minimize most of the aberrations. In the simulations the POF is represented by an area light source, while the grating is analyzed for different orders and the highest possible efficiency. In general, the element should be designed in a way that it can be produced with a mass production technology like injection molding in order to offer a reasonable price. However, designing the elements with regard to injection molding leads to some inherent challenges. The microstructure of an optical grating and the thick-walled 3D molded parts both result in high demands on the injection molding process. This also requires complex machining of the molding tool. Therefore, different experiments are done to optimize the process parameter, find the best molding material, and find a suitable machining method for the molding tool. The paper will describe the development of the demultiplexer by means of ray-tracing simulations step by step. Also, the process steps and the realized solutions for the injection molding are described.


Detection of p53 in Hodgkin's disease using the monoclonal antibody PAb248.
The recent demonstration that the murine anti-p53 monoclonal antibody PAb248 can identify human p53 in a variety of normal tissues proves that immunohistochemical detection does not necessarily indicate the presence of mutations. PAb248 can detect p53 protein in a cytoplasmic-perinuclear localization, not previously described. The present study presents the expression of this antibody in a series of 34 cases of Hodgkin's disease, comparing it with the antibodies CM1, PAb1801, and PAb240. In all cases, PAb248 showed uniform cytoplasmic-perinuclear staining in small and medium-sized lymphocytes, while it was constantly negative in Hodgkin, Reed-Sternberg (R-S/H) cells, and variants. This pattern of staining was the opposite to that observed with the antibodies CM1, PAb1801, and PAb240, where the staining was nuclear and restricted to the R-S/H cells, with the small lymphocytes being negative. p53 can be found in different conformations and localizations, with the cytoplasmic-perinuclear localization mainly, although not exclusively, being found in normal and reactive tissues and the nuclear localization being mainly expressed by neoplastic cells. These results give further support to the theory that the R-S/H cells are the neoplastic population in Hodgkin's disease, while the surrounding lymphocytes are reactive.