Thoughts on the end of life in patients with oxygen-dependent chronic obstructive pulmonary disease: A qualitative interview study.
The aim of the study was to deepen the current knowledge of how patients with chronic obstructive pulmonary disease and long-term oxygen treatment think about and expect end-of-life. A qualitative design was used. A purposeful sample of 19 patients with oxygen-dependent chronic obstructive pulmonary disease was obtained from the Swedish National Registry on Respiratory Failure (Swedevox). Data was collected with semi-structured interviews and analysed using a hermeneutic approach. The analysis revealed three themes: Living in the present without a future; difficulty talking about the uncertainty; and feeling anxious about leaving family behind. Participants indicated that healthcare professionals should invite them to mutual discussions as it was easier to reject an invitation if they could not talk right then, than to initiate a discussion themselves. Start of home oxygen or a deteriorating health status may be an important time to clinically address existential and end-of-life issues.


Modernizing problem formulation for risk assessment necessitates articulation of mode of action.
The process of scientific hypothesis formulation affects the experimental designs, methods and interpretations applied, but to be testable, the hypotheses posed must conform to the state of scientific knowledge and available technology. An analogous situation exists in risk assessment, where the questions addressed are typically articulated in the problem formulation phase. Decades ago, regulatory agencies couched problem formulation according to the questions answerable by the science of the day. As regulatory requirements for risk assessment became codified, so too did the rudiments of problem formulation. Unfortunately, codifying problem formulation prevented it from evolving to keep pace with scientific advancements. Today's more advanced science is not always being used effectively and efficiently in risk assessment because the risk assessment problem formulation step still typically poses antiquated questions. Problem formulation needs to be improved so that modern science can inform risk considerations. Based on recent developments in the Human Relevance Framework and using well-studied example chemicals - chloroform and carbon tetrachloride - an approach is proposed for focusing problem formulation on human-relevant hypotheses. We contend that modernizing problem formulation in this way will make risk assessment more scientifically accurate, more practical, and more relevant for protecting human health and the environment.


The effect of adding kinesiotaping versus pelvic floor exercise to conventional therapy in the management of post-colonoscopy coccydynia: a single-blind randomized controlled trial.
Coccydynia is a challenging disorder that is frequently managed conservatively. This study aimed to evaluate the efficacy of adding kinesiotaping versus pelvic floor exercise to conventional therapy in the management of post-colonoscopy coccydynia. Forty-two participants, aged 25-45 years, were randomly assigned to: the conventional therapy group (CT) receiving Piriformis and Iliopsoas muscle stretching exercise, clamshell exercise, and seat cushioning; the CT plus kinesiotaping group (CT-KT) receiving additional kinesiotaping; or the CT plus pelvic floor exercise (PFE) group (CT-PFE) executing additional PFE. All groups completed 4 weeks of training, 3 days a week. Pain intensity, assessed by the Pain Numeric Rating Scale (PNRS), and functional disability, evaluated by the Oswestry Disability Index (ODI), were estimated at baseline and after 4 weeks. There were significant inter-group variations in PNRS and ODI, where (P = 0.0001) and (P = 0.03), respectively. Differences between experimental groups were noteworthy in terms of NPRS, where the major change was in favor of group CT-KT (P = 0.001). However, there was no significant difference between them regarding their impact on ODI. CT-KT is more effective than CT-PFE in reducing pain associated with coccydynia post-colonoscopy, but there is no difference in their effects on functional disability. CT-KT is therefore recommended as an alternative treatment option for post-colonoscopy coccydynia.


Predictors of impaired bone health in long-term survivors after allogeneic stem cell transplantation.
Survival after allogeneic stem cell transplantation (allo-HSCT) has improved, but so have long-term sequelae. We studied risk factors for fractures and impaired bone health in allo-HSCT patients in the Basel HSCT registry from 01/2003 to 12/2014 using cox proportional models adjusted for age, gender and Karnofsky Index. Our primary endpoint was the incidence of fractures. Out of 652 patients, 32 (5.0%) had a new fracture after transplantation (yearly incidence rate of 1.6%, 95% Confidence Interval [95%CI] 1.1-2.3%) and 325 (49.8%) had low bone mineral density (yearly incidence rate of 13.1%, 95%CI 11.6-14.8%), including 36.0% with osteopenia and 13.8% with osteoporosis. We found vitamin D deficiency during follow-up (Hazard Ratio [HR] 1.25, 95%CI 1.11-1.41, p < 0.001), hyperthyroidism before transplantation (HR 4.85, 95%CI 1.05-22.54, p = 0.044), cumulative years of immunosuppressant exposure (HR 1.23, 95%CI 1.07-1.41, p = 0.004 for steroidal and HR 1.09, 95%CI 1.01-1.18, p = 0.025 for non-steroidal drugs) and graft-versus-host disease (acute HR 1.24, 95%CI 1.11-1.40, p < 0.001; chronic HR 2.82, 95%CI 1.12-7.13, p = 0.028) to be significantly associated with fractures. Patients undergoing HSCT are at increased risk of fractures, which is associated with various disease and treatment-specific factors. Early identification of patients at risk may help to improve preventive measures.


On the sub-maximal yield and photo-electric stimulation of chlorophyll a fluorescence in single turnover excitations in plant cells.
A set of expressions is derived which quantifies the chlorophyll fluorescence yield in terms of rate constants of primary light reactions of PSII, the fraction of open and semi-open RCs and of the electric field sensed by the RC in the thylakoid membrane. The decay kinetics of the chlorophyll fluorescence yield after a single turnover excitation in the presence of DCMU show at least two components, one reversible within approx. 1 s and one with a dark reversion lasting more than 30 s. The latter is attributed to photochemical quenching; the fast component is interpreted to be associated at least partially with photo-electrochemical control. It will be illustrated that (i) the sub-maximal fluorescence yield in single turnover excitation is associated with semi-closure of RCs, (ii) the trapping efficiency of semi-closed centers is less than 50% of that of open centers and (iii) the fluorescence yield of antennas with semi-closed RCs has the highest sensitivity to changes in strength of photo-electric fields.


Fabrication of biomimetic high performance antireflective and antifogging film by spin-coating.
In this paper, we report a facile way to fabricate biomimetic high performance optical hybrid films with excellent antireflective and antifogging properties by one-step spin-coating the mixture of mesoporous SiO(2) particles and SiO(2) sol. The production process of the films is easy, low-cost, and time-efficient. Mesoporous SiO(2) particles containing surfactants disperse in SiO(2) sol stably without any chemical modification, which decrease the effective refractive index and increase the transmittance of the films. In addition, such films possess superhydrophilic properties and exhibit high performance antifogging properties. Due to the good film forming performance of SiO(2) sol, mesoporous SiO(2) particles are embedded in the films and impart the films high mechanical stability and durability. The surface morphology of the films can maintain well after repeated friction, and the performances of antireflective and antifogging also do not change as well.


Glucagon secretion determined by the RIA method is lower in patients with low left ventricular ejection fraction: The new glass study.
We investigated the glucagon levels in patients with heart failure (HF), using long oral glucose tolerance test (OGTT). In this prospective observational study, we enrolled 30 undiagnosed diabetes patients (age 69 ± 10 years, 70% males, HbA1c 43 mmol/mol). A 4-h OGTT was performed. Glucose, insulin, and glucagon (radioimmunoassay [RIA] and sandwich ELISA [S-W] methods) were evaluated during 4-h. We compared glucagon levels between HF and non-HF patients. There were 11 HF and 19 non-HF patients. In patients with HF, glucagon (S-W) during 4-h was lower than in patients without HF, with no significant difference. The area under the curve (AUC) of glucagon (RIA) during 4-h was significantly lower among HF patients. Moreover, in patients with reduced left ventricular ejection fraction (LVEF) (<40%), AUC glucagon (RIA) was significantly lower than in patients with non-reduced EF (≥40%). However, there was no difference in glucagon values between the high E/e' (≥13.0) and low E/e' (<13.0) groups. Although glucagon (S-W) showed no significant difference in patients with and without HF, especially reduced LVEF, glucagon (RIA) secretion was significantly lower in HF patients than in patients without HF. It is suggested that low glucagon secretion might be correlated with low EF.


A nonparametric instrumental approach to confounding in competing risks models.
This paper discusses nonparametric identification and estimation of the causal effect of a treatment in the presence of confounding, competing risks and random right-censoring. Our identification strategy is based on an instrumental variable. We show that the competing risks model generates a nonparametric quantile instrumental regression problem. Quantile treatment effects on the subdistribution function can be recovered from the regression function. A distinguishing feature of the model is that censoring and competing risks prevent identification at some quantiles. We characterize the set of quantiles for which exact identification is possible and give partial identification results for other quantiles. We outline an estimation procedure and discuss its properties. The finite sample performance of the estimator is evaluated through simulations. We apply the proposed method to the Health Insurance Plan of Greater New York experiment.


The plant cytolytic protein enterolobin assumes a dimeric structure in solution.
Enterolobin is a plant cytolytic protein similar to the bacterial cytolysin aerolysin. Biochemical and biophysical techniques were used to verify if enterolobin, like aerolysin, adopts a dimeric structure in solution. SDS-PAGE showed bands corresponding to enterolobin monomer, dimer and oligomers, whilst gel filtration chromatography and electrospray mass spectrometry revealed preferred association of enterolobin as a dimer. Atomic force microscopy (AFM) of enterolobin showed images of a dimer assembly at a concentration as low as 10 microg/ml, similarly to aerolysin. The enterolobin in silico docked structure is coherent with AFM enterolobin dimer shapes.


Immunogenicity correlation in cynomolgus monkeys between Luminex-based total IgG immunoassay and pseudovirion-based neutralization assay for a 14-valent recombinant human papillomavirus vaccine.
A new virus-like particle based vaccine covering 14 types of high-risk and disease-inducing human papillomavirus (HPV) can offer better coverage against HPV-induced diseases, particularly cervical cancers. However, the assessment of immunogenicity of the vaccine is an important task, representing not only its significant clinical characteristics, but also a major challenge, in terms of both the suitability of methods and the clinical sample testing throughput supporting clinical development. This work covers the development and evaluation of a method based on Luminex technology (a coded-bead and flow-cytometric approach) to assess the HPV-type specific total immunoglobulin G (IgG). This method can evaluate the antibodies in sera post immunization against multiple types of HPV simultaneously (i.e., with multiplexing capability), save time and cost, and improve test throughput with higher sensitivity and precision than the classical, plate-based enzyme-linked immunoassay and competitive Luminex-based immunoassays. Using cynomolgus monkeys as model, we demonstrated the good correlation between the results from the pseudovirion-based neutralization assay (PBNA), and the Luminex-based total IgG assay, supporting that the latter method can be considered as a viable, dependable replacement method for the PBNA supporting immunogenicity evaluation of HPV vaccine in preclinical development and clinical investigation.


A comparison of angiotensinase and placental leucine aminopeptidase during normal pregnancy.
1. Serum leucine aminopeptidase activity measured by its action on hydrolysis of L-leucyl-beta-naphthylamide was increased progressively as pregnancy advanced. Angiotensinase activity determined by bioasaay showed parallel increase during normal pregnancy. 2. The effects of heat treatment and L-methionine on the activity of angiotensinase were compared with those on leucine aminopeptidase activity in pregnancy serum.


Problem and pathological gambling are associated with poorer mental and physical health in older adults.
To evaluate the prevalence and correlates of problem and pathological gambling in older adults. Adults (n = 343) aged 60 years and older attending senior centers, bingo sites and other community activities completed a screening form containing the South Oaks Gambling Screen and the Short Form-12 Health Survey, to evaluate physical and mental health. Overall, 6.4% of the respondents were classified as problem gamblers and an additional 3.8% as pathological gamblers. Problem and pathological gamblers evidenced significantly greater physical and mental health problems than non-problem gamblers. These data suggest that about 10 percent of active older adults experience gambling problems, which are associated with poor physical and mental health.


Bilateral coronoid hyperplasia. Report of a case.
A case of bilateral coronoid hyperplasia is presented. This is a developmental abnormality that occurs primarily in males and is characterized by progressive, asymptomatic limitation of mandibular movements. The condition is usually detected shortly after the age of puberty. A diagnosis is usually made on the basis of the history and the radiographic findings. Surgical removal of the elongated coronoid processes, along with good postoperative range-of-motion exercises, will restore full mandibular movement.


Enalapril clinical pharmacokinetics and pharmacokinetic-pharmacodynamic relationships. An overview.
The conventional pharmacokinetic profile of the angiotensin converting enzyme (ACE) inhibitor, enalapril, is a lipid-soluble and relatively inactive prodrug with good oral absorption (60 to 70%), a rapid peak plasma concentration (1 hour) and rapid clearance (undetectable by 4 hours) by de-esterification in the liver to a primary active diacid metabolite, enalaprilat. Peak plasma enalaprilat concentrations occur 2 to 4 hours after oral enalapril administration. Elimination thereafter is biphasic, with an initial phase which reflects renal filtration (elimination half-life 2 to 6 hours) and a subsequent prolonged phase (elimination half-life 36 hours), the latter representing equilibration of drug from tissue distribution sites. The prolonged phase does not contribute to drug accumulation on repeated administration but is thought to be of pharmacological significance in mediating drug effects. Renal impairment [particularly creatinine clearance < 20 ml/min (< 1.2 L/h)] results in significant accumulation of enalaprilat and necessitates dosage reduction. Accumulation is probably the cause of reduced elimination in healthy elderly individuals and in patients with concomitant diabetes, hypertension and heart failure. Conventional pharmacokinetic approaches have recently been extended by more detailed descriptions of the nonlinear binding of enalaprilat to ACE in plasma and tissue sites. As a result of these new approaches, there have been significant improvements in the characterisation of concentration-time profiles for single-dose administration and the translation to steady-state. Such improvements have further importance for the accurate integration of the pharmacokinetic and pharmacodynamic responses to enalapril(at) in a concentration-effect model.(ABSTRACT TRUNCATED AT 250 WORDS)


A novel siderophore system is essential for the growth of Pseudomonas aeruginosa in airway mucus.
Pseudomonas aeruginosa establishes airway infections in Cystic Fibrosis patients. Here, we investigate the molecular interactions between P. aeruginosa and airway mucus secretions (AMS) derived from the primary cultures of normal human tracheal epithelial (NHTE) cells. PAO1, a prototype strain of P. aeruginosa, was capable of proliferating during incubation with AMS, while all other tested bacterial species perished. A PAO1 mutant lacking PA4834 gene became susceptible to AMS treatment. The ΔPA4834 mutant was grown in AMS supplemented with 100 μM ferric iron, suggesting that the PA4834 gene product is involved in iron metabolism. Consistently, intracellular iron content was decreased in the mutant, but not in PAO1 after the AMS treatment. Importantly, a PAO1 mutant unable to produce both pyoverdine and pyochelin remained viable, suggesting that these two major siderophore molecules are dispensable for maintaining viability during incubation with AMS. The ΔPA4834 mutant was regrown in AMS amended with 100 μM nicotianamine, a phytosiderophore whose production is predicted to be mediated by the PA4836 gene. Infectivity of the ΔPA4834 mutant was also significantly compromised in vivo. Together, our results identify a genetic element encoding a novel iron acquisition system that plays a previously undiscovered role in P. aeruginosa airway infection.


Regional specification of rodent and human neurospheres.
Neural precursor cells were isolated from various regions of the developing rat and human brain and grown in culture as aggregates termed neurospheres. We asked whether cells within human and rodent neurospheres are identical, or whether they have species specific characteristics or differences based on their region of origin. Under our culture conditions, rodent neurospheres isolated from the cortex (ctxNS) and striatum (strNS) grew faster than those from the mesencephalon (mesNS), but stopped growing after only eight to ten population doublings. In contrast, human neurospheres under identical culture conditions, continued to grow for over 40 population doublings. Following migration and differentiation of both rodent and human cultures, ctxNS and strNS generated high numbers of small neurons whereas mesNS generated small numbers of large neurons with many long fibres. Only very rare neurons from mesNS expressed dopaminergic markers, and thus may require further signals to fully mature. While the rat neurospheres generated high numbers of oligodendrocytes, very few were found to develop from human neurospheres from any region after a few weeks of passaging. FACS analysis revealed a unique population of smaller cells within human strNS and ctxNS, which appeared to be neuronal progenitors. However, large cells within neurospheres were capable of generating these small neuronal progenitors following further proliferation. Together, our data show that rat and human neurospheres have unique characteristics with regard to growth and differentiation, and that the majority of precursor cells within neurospheres are regionally specified to generate set numbers of neurons. These findings have important implications for understanding the nature of proliferating neural precursors isolated from the developing CNS, and their potential for brain repair.


Sources of transcription variation in Plasmodium falciparum.
Variation in transcript abundance can contribute to both short-term environmental response and long-term evolutionary adaptation. Most studies are designed to assess differences in mean transcription levels and do not consider other potentially important and confounding sources of transcriptional variation. Detailed quantification of variation sources will improve our ability to detect and identify the mechanisms that contribute to genome-wide transcription changes that underpin adaptive responses. To quantify innate levels of expression variation, we measured mRNA levels for more than 5000 genes in the malaria parasite, Plasmodium falciparum, among clones derived from two parasite strains across biologically and experimentally replicated batches. Using a mixed effects model, we partitioned the total variation among four sources-between strain, within strain, environmental batch effects, and stochastic noise. We found 646 genes with significant variation attributable to at least one of these sources. These genes were categorized by their predominant variation source and further examined using gene ontology enrichment analysis to associate function with each source of variation. Genes with environmental batch effect and within strain transcript variation may contribute to phenotypic plasticity, while genes with between strain variation may contribute to adaptive responses and processes that lead to parasite strain-specific survival under varied conditions.


Pathophysiological and diagnostic importance of fatty acid-binding protein 1 in heart failure with preserved ejection fraction.
Elevated intracardiac pressure at rest and/or exercise is a fundamental abnormality in heart failure with preserved ejection fraction (HFpEF). Fatty acid-binding protein 1 (FABP1) is proposed to be a sensitive biomarker for liver injury. We sought to determine whether FABP1 at rest would be elevated in HFpEF and would correlate with echocardiographic markers of intracardiac pressures at rest and during exercise. In this prospective study, subjects with HFpEF (n = 22) and control subjects without HF (n = 23) underwent resting FABP1 measurements and supine bicycle exercise echocardiography. Although levels of conventional hepatic enzymes were similar between groups, FABP1 levels were elevated in HFpEF compared to controls (45 [25-68] vs. 18 [14-24] ng/mL, p = 0.0008). FABP1 levels were correlated with radiographic and blood-based markers of congestion, hemodynamic derangements during peak exercise (E/e', r = 0.50; right atrial pressure, r = 0.35; pulmonary artery systolic pressure, r = 0.46), reduced exercise cardiac output (r = - 0.49), and poor exercise workload achieved (r = - 0.40, all p < 0.05). FABP1 distinguished HFpEF from controls with an area under the curve of 0.79 (p = 0.003) and had an incremental diagnostic value over the H2FPEF score (p = 0.007). In conclusion, FABP1 could be a novel hepatic biomarker that associates with hemodynamic derangements, reduced cardiac output, and poor exercise capacity in HFpEF.


EEG development of healthy boys and girls. Results of a longitudinal study.
This longitudinal study has extended an earlier cross-sectional one in order to investigate the development of background activity and the topographical distribution of different EEG parameters during childhood and adolescence. Ninety-six healthy children (47 boys and 49 girls) were followed for up to 7 years with serial EEGs. These were analysed by spectral analysis. Power spectra were calculated for conventional frequency bands. Three topics were followed: (i) differentiation between inter-individual and age-related changes, (ii) evaluation of sex differences and (iii) evaluation of developmental velocity of different EEG parameters. Between 4 and 17 years the content of theta waves decreases and the content of alpha waves increases. In girls more than in boys, slow alpha waves (7.5-9.5 c/sec) in occipital leads first increase, as theta decreases. Later they decrease themselves while fast alpha waves (9.5-12.5 c/sec) increase. The velocity of occipital changes is almost twice that centrally, and is greater in early childhood than after 10 years of age. Girls under 6 years of age show significantly more theta and less alpha waves compared with boys. However, the developmental velocity (change per year) is higher in girls than in boys and continues for a longer time; thus girls catch up with age-related changes of the boys by a later developmental spurt. Individual characteristics of the subjects determine the EEG twice as much as the age factor; this explains the wide range of results in previous studies. No trend was found to suggest earlier maturation in one hemisphere while in 5% of the subjects the amount of activity for one parameter in one hemisphere exceeds 1.5 times the amount of activity in the other. The individual characteristics of the EEG spectra show a remarkable stability over the years.


Parenting a newborn baby during the COVID-19 pandemic: a qualitative survey.
The COVID-19 pandemic caused long periods of lockdown, social isolation and intense challenges for parents. This study examines parenting in an infant cohort born at the pandemic onset. The CORAL study is a prospective longitudinal observational study looking at allergy, immune function and neurodevelopmental outcome in babies born between March and May 2020. Demographic information was collected, babies were reviewed at 6-monthly intervals, and serology for COVID-19 infection was recorded. When babies were 12 months old, parents were asked for 3-5 words to describe raising a baby during the pandemic. Frequency of word usage was compared between first time parents and parents with other children, and parents of babies with and without a diagnosis of COVID-19 infection. 354 babies were recruited to CORAL study. Social circles were small. At 6 months the median number of people (including parents) who had kissed the baby was 3, and by 12 months one-quarter of babies had never met another child of similar age. 304 parents completed the word choice. Commonly reported words were lonely (44.4%), isolating (31.9%) and strong bond (15.8%). 12 of those 304 babies had COVID-19 in their first year of life and there was no significant difference in reported negative or positive word number compared with parents of babies without a COVID-19 infection, or by first time parents or those who already had children. The lockdowns and social restrictions made raising an infant challenging for all parents in Ireland. It is important parents know this was a shared experience.


Inhibitory effect of pentagastrin and feeding natural and motilin-induced interdigestive contractions in the stomach of conscious dogs.
The interaction between motilin, pentagastrin and feeding was investigated during the interdigestive gastric motor state in 4 healthy conscious dogs. We confirmed previous reports that an i.v. infusion of synthetic motilin always induced a pattern precisely like that of the naturally-occurring interdigestive contractions of the stomach. Pentagastrin inhibited the occurrence of the interdigestive contractions at dose between 0.2-1.6 microgram/kg-hr; this inhibition had a dose-related relationship to the occurrence of the motilin-induced contractions. The ingestion of food strongly inhibited the occurrence of not only the natually-occurring contractions but also the motilin-induced contractions in the stomach and changed the motor pattern to the digestive one. These findings strongly support our hypothesis that the interdigestive contractions in the dog are, at least in part, under the control of motilin. Furthermore, it should be emphasized that pentagastrin not always stimulates gastric motor activity but conversely, in the interdigestive state it inhibits movements characteristic of that condition and replaces them with the digestive pattern.


Endovascular vs. open surgery of abdominal aortic aneurysm in high-risk patients: a single center experience.
We compared two groups of high-risk patients with abdominal aortic aneurysm to assess the safety and efficacy of endovascular repair vs. open surgery. From January 1998 to July 2003, sixty-two high-risk patients were divided into two groups: group A consisted of 28 (46 %) open surgery patients and group B consisted of 34 (54 %) patients who underwent endovascular repair. Four patients (14.3 %) in the open surgery group died, while no deaths occurred in the endovascular group ( p < 0.05). There were 14 complications in 8 patients of the open surgery group versus 2 complications in 2 patients of the endovascular group ( p = 0.01). At follow-up there were 4 (16.6 %) deaths in group A and 3 (8.8 %) in the endovascular group ( p = n. s.). While the use of endovascular repair in patients who are physiologically fit for open surgical repair remains controversial, we believe that patients with multiple or advanced comorbidities, i.e. high-risk patients, can benefit from the endografting procedure.


Myosin dynamics during relaxation in mouse soleus muscle and modulation by 2'-deoxy-ATP.
Skeletal muscle relaxation has been primarily studied by assessing the kinetics of force decay. Little is known about the resultant dynamics of structural changes in myosin heads during relaxation. The naturally occurring nucleotide 2-deoxy-ATP (dATP) is a myosin activator that enhances cross-bridge binding and kinetics. X-ray diffraction data indicate that with elevated dATP, myosin heads were extended closer to actin in relaxed muscle and myosin heads return to an ordered, resting state after contraction more quickly. Molecular dynamics simulations of post-powerstroke myosin suggest that dATP induces structural changes in myosin heads that increase the surface area of the actin-binding regions promoting myosin interaction with actin, which could explain the observed delays in the onset of relaxation. This study of the dATP-induced changes in myosin may be instructive for determining the structural changes desired for other potential myosin-targeted molecular compounds to treat muscle diseases. Here we used time-resolved small-angle X-ray diffraction coupled with force measurements to study the structural changes in FVB mouse skeletal muscle sarcomeres during relaxation after tetanus contraction. To estimate the rate of myosin deactivation, we followed the rate of the intensity recovery of the first-order myosin layer line (MLL1) and restoration of the resting spacing of the third and sixth order of meridional reflection (SM3 and SM6 ) following tetanic contraction. A transgenic mouse model with elevated skeletal muscle 2-deoxy-ATP (dATP) was used to study how myosin activators may affect soleus muscle relaxation. X-ray diffraction evidence indicates that with elevated dATP, myosin heads were extended closer to actin in resting muscle. Following contraction, there is a slight but significant delay in the decay of force relative to WT muscle while the return of myosin heads to an ordered resting state was initially slower, then became more rapid than in WT muscle. Molecular dynamics simulations of post-powerstroke myosin suggest that dATP induces structural changes in myosin that increase the surface area of the actin-binding regions, promoting myosin interaction with actin. With dATP, myosin heads may remain in an activated state near the thin filaments following relaxation, accounting for the delay in force decay and the initial delay in recovery of resting head configuration, and this could facilitate subsequent contractions.


A dynamic physical-distancing model to evaluate spatial measures for prevention of Covid-19 spread.
Motivated by the global pandemic of COVID-19, this study investigates the spatial factors influencing physical distancing, and how these affect the transmission of the SARS-CoV-2 virus, by integrating pedestrian dynamics with a modified susceptible-exposed-infectious model. Contacts between infected and susceptible pedestrians are examined by determining physical-distancing pedestrian dynamics in three types of spaces, and used to estimate the proportion of newly infected pedestrians in these spaces. Desired behaviour for physical distancing can be observed from simulation results, and aggregated simulation findings reveal that certain layouts enable physical distancing to reduce the transmission of SARS-CoV-2. We also provide policymakers with several design guidelines on how to proactively design more effective and resilient space layouts in the context of pandemics to keep low transmission risks while maintaining a high pedestrian volume. This approach has enormous application potential for other infectious-disease transmission and space assessments.


A modified gel filtration technique producing an unusual exclusion volume of IgM: a simple way of preparing monoclonal IgM.
The vast majority of monoclonal IgM proteins is eluted just before the total volume of the column when filtered through G200 Sephadex or S200 Sephacryl gels equilibrated in a 0.005 M phosphate buffer but eluted with 0.05 M phosphate buffer containing 1.7 M NaCl. This unusual behaviour in low-ionic buffer is probably due to the poor solubility of IgM in diluted buffers. It allows a 1-step purification procedure under mild conditions and is suitable for both large and small scale preparations.


Does patient assessment of the quality of the primary care they receive predict subsequent outcomes? An Oklahoma Physicians Resource/Research Network (OKPRN) study.
We analyzed data from a cohort of 782 older patients assembled in 1999 to 2000 to determine whether baseline patient assessments of the quality of the primary care services they had received, measured using the Components of Primary Care Index (CPCI), were associated with subsequent changes in health-related quality of life (HRQoL) and/or survival. Longitudinal growth curve models were used to analyze changes in Quality of Well-Being (QWB-SA) scores over an average (S.D.) of 2.07 (1.07) years. Cox proportional hazards models were used to identify variables associated with mortality over an average of 8.26 years (6460 person-years). To reduce confounding by severity of illness, subjects were stratified into disabled, nondisabled high utilizers of primary care, and nondisabled low utilizers. Within subgroups, we controlled for number of chronic illnesses and scores on the General Health subscale of the Medical Outcomes Study Short Form-36 (SF-36). We also controlled for baseline age, sex, marital status, income, body mass index, educational attainment, duration of the relationship with current primary care physician (PCP), and number of visits to the PCP in the year before enrollment. Analyses took into account clustering of patients within PCP. Neither total CPCI nor any CPCI subscale score was associated with QWB-SA change over time. Higher ratings of Coordination of Care were associated with reduced survival in the disabled and nondisabled high utilizer subgroups (P = .007). Assuming that effective primary care results in better HRQoL and longer survival and that the CPCI captures important primary care attributes, older patients' level of satisfaction with the quality of their primary care may not be a good surrogate measure of effectiveness.


Enantiospecific synthesis and cytotoxicity of 7-(4-methoxyphenyl)-6-phenyl-2,3,8,8a-tetrahydroindolizin-5(1H)-one enantiomers.
An enantiospecific synthesis was developed to generate both enantiomers of 7-(4-methoxyphenyl)-6-phenyl-2,3,8,8a-tetrahydroindolizin-5(1H)-one. A biological assay utilizing the HCT-116 colon cancer cell line to determine the cytotoxicity of these analogs revealed that only the (R)-enantiomer exhibited appreciable cytotoxicity with an IC(50) value of 0.2 microM.


The index of orthognathic functional treatment need accurately prioritises those patients already selected for orthognathic surgery within the NHS.
The index of orthognathic functional treatment need (IOFTN) is a newly-proposed system to help to prioritise patients for orthognathic treatment. The five categories are similar to those used in orthodontics, but include additional parameters such as sleep apnoea and facial asymmetry. The aim of this audit was to validate the index and find out the potential future implications, should such a system ever be adopted by commissioners. We calculated the IOFTN category of 100 consecutive patients who had orthognathic surgery between 2010-14 using clinical notes, photographs, study models, and radiographs, and determined the number in categories 4 or 5, analogous to the current indications for orthodontic treatment within the NHS. Sufficient clinical information was available to categorise 59/100 patients, and 56 of the 59 (95%) were in either category 4 or 5. All three of the remaining patients (in categories 1-3) who were operated on were treated because of the anticipated favourable impact on their quality of life. The IOFTN has been proposed for use in future commissioning of orthognathic services within the NHS, and this study has confirmed its efficacy in prioritising treatment accurately, with 95% of patients being in categories 4 or 5. We recommend that the orthognathic treatment index be adapted to include additional psychosocial assessment so that patients who fall into the lower functional categories are not automatically excluded from this potentially life-changing treatment.


Identification and partial characterization of three calcium- and zinc-independent gelatinases constitutively present in human circulation.
Three constitutive gelatinases in human plasma were identified and characterized relative to known matrix metalloproteinase (MMP) gelatinases: MMP-2 (fibroblast 72-kDa) and MMP-9 (neutrophil 92-, 130-, and 225-kDa). Substrate gel electrophoresis (gelatin zymography) revealed an apparent Mw of 78-, 82-, and 89-kDa for these gelatinases. Densitometry revealed that MMP-9 and MMP-2 were highly calcium sensitive requiring 50-150 microM and 500 microM calcium for half-maximal activity, respectively. Of the new gelatinases, only the 89-kDa form demonstrated slight calcium activation. The three gelatinases were unaffected by known MMP inhibitors: EDTA (5 mM), 1,10-phenanthroline (2 mM), and pepstatin (18 microM). Serine and thiol protease inhibitors (leupeptin, aprotinin, PMSF, TLCK, TPCK, antichymostatin, antipain) were also ineffective. Solution-phase IEF revealed that the 78- and 82-kDa forms focused at neutral pI 6.72-7.95 whereas the 89-kDa focused at an acidic pI 4.89-5.18 (similar to neutrophil and fibroblast forms). The data indicate that these gelatinases are not MMPs or partially activated MMPs. Their role in normal and pathological conditions is not known.


Orexins, a novel family of hypothalamic neuropeptides, modulate pituitary luteinizing hormone secretion in an ovarian steroid-dependent manner.
Orexin A and orexin B are a newly described family of orexigenic hypothalamic neuropeptides. The distribution of orexin immunoreactive fibers overlaps with the luteinizing hormone-releasing hormone (LHRH) neuronal system in the septo-preoptic area and the arcuate nucleus-median eminence region. To test the hypothesis that orexins may be involved in the regulation of pituitary LH secretion by influencing LHRH release, the effects of orexin A and orexin B on LH secretion were evaluated in ovariectomized (ovx) and ovarian steroid-treated ovx rats. Intracerebroventricular injection of orexin A or orexin B rapidly stimulated LH secretion in a dose- (0.3 and 3.0 nmol) and time-related fashion in estradiol benzoate- (EB) and progesterone (P)-pretreated ovx rats. Peak plasma LH concentration at 10 min were significantly higher in orexin A- than in orexin B-injected rats. On the other hand, both orexins (3 nmol) inhibited LH release in unprimed ovx rats. This ovarian steroid-dependent bimodal LH response is reminiscent of that of other orexigenic neuropeptides, such as neuropeptide Y. These findings show that orexins belong to a group of hypothalamic messengers that neurochemically bridge the regulatory networks that controls reproduction and energy balance.


Elaboration and controlling excited state double proton transfer mechanism of 2,5-bis(benzoxazol-2-yl)thiophene-3,4-diol.
In the present work, we theoretically illuminate the excited state double proton transfer (ESDPT) process about a novel synthesized system 2,5-bis(benzoxazol-2-yl)thiophene-3,4-diol (BBTD). Minor changes of both structure and charge redistribution deriving from photoexcitation result in obviously different excited state dynamical process. Exploration about our constructed S1-state potential energy surface (PES) indicates a stepwise ESDPT mechanism for BBTD. In addition, we present a new mechanism about regulating and controlling stepwise ESDPT process via external electric field.


Basic mechanisms of tendon fatigue damage.
Pathologic processes intrinsic and extrinsic to the tendons have been proposed as the underlying cause of rotator cuff disease, but the precise etiology is not known. Tear formation is, in part, attributable to the accumulation of subrupture tendon fatigue damage. We review the molecular, mechanical, and structural changes induced in tendons subjected to controlled amounts of fatigue loading in an animal model of early tendinopathy. The distinct tendon responses to low and moderate levels of loading, as opposed to high levels, provide insight into the potential mechanisms for the therapeutic benefits of exercise in the treatment of rotator cuff tendinopathy. The progression of damage accumulation leading to fiber rupture and eventual tendon tearing seen with higher loading illustrates the progression from tendinopathy to full-thickness tearing. We hope that this more realistic animal model of tendon fatigue damage will allow better assessment of biologic, mechanical, tissue-engineering, and rehabilitation strategies to improve repair success.


Effects of viral exposure of the two-cell mouse embryo on cleavage and blastocyst formation in vitro.
The effect of viral exposure of two-cell mouse embryos on their capacity to undergo subsequent cleavage and blastocyst formation in vitro was determined. Exposure to Coxsackie viruses B-4 and B-6, reovirus type 2, influenza virus type A, mouse cytomegalovirus, adenovirus type 5, and mouse adenovirus resulted in statistically significant inhibition of blastocyst formation. Development in vitro was unaffected by exposure to ECHO virus type 11, attenuated poliomyelitis virus type 2, parainfluenza virus type 1, mumps, rubella, and herpes simplex viruses types 1 and 2. Blastocyst formation was also unaffected by exposure of embryos to mouse interferon in a concentration 24 units/ml of culture fluid. Coxsackie virus B-4 was recovered from exposed embryos.


Pyrodextrinization of yam (Dioscorea sp.) starch isolated from tubers grown in Brazil and physicochemical characterization of yellow pyrodextrins.
This study optimizes the pyrodextrinization of yam (Dioscorea sp.) starch isolated from tubers grown in Brazil to produce a yellow pyrodextrin with the lowest enzymatically available starch (AS) content and color difference (ΔE) index. At 140 °C (fixed heating temperature), the effects of acid concentration (0.65 - 2.99 g of HCl/kg of starch) and incubation time (53 - 307 min) on the response variables were evaluated using a response surface methodology. Some physicochemical characteristics were also determined on pyrodextrins. Both factors negatively affected the AS content, although positively influenced the ΔE (P < 0.05). The yellow pyrodextrin produced with 1.82 g/kg and heating for 307 min, presented physicochemical properties similar to the commercial pyrodextrins from potato starch, with 46.6 % of AS, 24.5 of ΔE, high solubility and very low viscosity. The pyrodextrinization caused a decrease of 30 - 54 % in AS content (P < 0.05), making these yam pyrodextrins a promising material for water-soluble and very low viscous dietary fiber.


A novel protein truncating mutation in L2HGDH causes L-2-hydroxyglutaric aciduria in a consanguineous Pakistani family.
L-2-hydroxyglutaric aciduria (L2HGA) is a rare neurometabolic disorder that occurs due to accumulation of L-2-hydroxyglutaric acid in the cerebrospinal fluid (CSF), plasma and urine. The clinical manifestation of L2HGA includes intellectual disability, cerebellar ataxia, epilepsy, speech problems and macrocephaly. In the present study, we ascertained a multigenerational consanguineous Pakistani family with 5 affected individuals. Clinical studies were performed through biochemical tests and brain CT scan. Locus mapping was carried out through genome-wide SNP genotyping, whole exome sequencing and Sanger sequencing. For in silico studies protein structural modeling and docking was done using I-TASSER, Cluspro and AutoDock VINA tools. Affected individuals presented with cognitive impairment, gait disturbance, speech difficulties and psychomotor delay. Radiologic analysis of a male patient revealed leukoaraiosis with hypoattenuation of cerebral white matter, suggestive of hypomyelination. Homozygosity mapping in this family revealed a linkage region on chromosome 14 between markers rs2039791 and rs781354. Subsequent whole exome analysis identified a novel frameshift mutation NM_024884.3:c.180delG, p.(Ala62Profs*24) in the second exon of L2HGDH. Sanger sequencing confirmed segregation of this mutation with the disease phenotype. The identification of the most N-terminal loss of function mutation published thus far further expands the mutational spectrum of L2HGDH.


Disruption of Col9a2 expression leads to defects in osteochondral homeostasis and osteoarthritis-like phenotype in mice.
As one of the branched chains of Type IX collagen (Col9), Collagen IX alpha2 (Col9a2) has been reported to be associated with several orthopedic conditions. However, the relationship between Col9a2 and knee osteoarthritis (KOA) remains to be elucidated. To probe the relationship between Col9a2 and KOA, we performed a systematic analysis of Col9a2-deficient (Col9a2-/-) mice using whole-mount skeletal staining, Micro-CT (μCT), biomechanics, histomorphometry, immunohistochemistry (IHC), immunofluorescence (IF) and Elisa. We found that the subchondral bone (SCB) in the knee joint of Col9a2-/- mice became sparse and deformed in the early stage, with altered bone morphometric parameters, reduced load-bearing capacity, dysfunctional bone homeostasis (decreased osteogenesis capacity and elevated bone resorption capacity), diminished cartilage proteoglycans and disrupted cartilage extracellular matrix (ECM) anabolism and catabolism compared with the Col9a2+/+ mice. In the late stage, the cartilage degeneration in Col9a2-/- mice were particularly pronounced compared to Col9a2+/+ mice, as evidenced by severe cartilage destruction and a marked reduction in cartilage thickness and area. Overall, Col9a2 is essential for maintaining osteochondral homeostasis in the knee joint of mice, and the absence of this gene is accompanied by distinct sclerosis of the SCB and a reduction in load-bearing capacity; in the late stage, in the lack of SCB stress inhibition, excessive load is consistently exerted on the cartilage, ultimately leading to osteoarthritic-like articular cartilage damage.


Trapeziectomy with suture-button suspensionplasty versus ligament reconstruction and tendon interposition: a randomized controlled trial.
The purpose of the present study was to compare the results of patients operated with trapeziectomy and ligament reconstruction and tendon interposition (LRTI) using flexor carpi radialis tendon versus trapeziectomy followed by suspension of the first metacarpal to the second metacarpal using a Mini TightRope® suture button (suture button suspension: SBS). A single-center prospective randomized controlled trial was performed, comparing 37 patients with SBS and 39 with LRTI. All surgeries were performed by the same fellowship-trained hand surgeon. Patients were assessed by an independent observer at 40 months' follow-up. Pre- and postoperative strength, trapezial space ratio (TSR), range of motion, QuickDASH and visual analogue pain score were recorded. Both procedures improved functional parameters of pain, key strength, tip strength and grip strength while maintaining range of motion, without significant differences. In the SBS group, TSR decreased by 17%, compared to 28% in the LRTI group. The mean operative time was shorter in SBS (63 vs 91 minutes; p < 0.0001), as was immobilization time (2 vs 6 weeks; p < 0.0001), and patients resumed normal activity sooner (10 vs 12 week; p = 0.0138) and required less physical therapy (19.3 vs 13.1 weeks; p < 0.0001). We believe that our results are related to the hypothesis suggested by biomechanical studies that revealed better initial load bearing profile and maintenance of trapezial space following serial loading in cadaver models.


Prolonged isolated thrombocytopenia after hematopoietic stem cell transplantation: morphologic correlation.
Prolonged isolated thrombocytopenia, defined as recovery of other cell counts with continuous dependence on platelet transfusions for greater than 90 days after hematopoietic stem cell transplantation (HSCT), develops in approximately 5% of patients who undergo HSCT. Although the clinical conditions associated with prolonged isolated thrombocytopenia have been studied, a systematic review of bone marrow biopsies has not been performed and the pathophysiologic basis has not been defined. We reviewed all HSCT at one center from 1990 to 1995 (n = 454) and found 12 cases that met criteria for prolonged isolated thrombocytopenia (incidence = 12/454 or 3%). Bone marrow core biopsies from 12 patients with prolonged isolated thrombocytopenia were reviewed to determine cellularity, numbers of megakaryocytes, the presence of atypical forms, and clusters of megakaryocytes. These marrow megakaryocyte counts were compared to age and disease matched controls, and 11 normal donors. Patients (aged 1-56 years, mean 32 years) who underwent HSCT (four sibling HLA-identical, five autologous bone marrow, three autologous peripheral stem cell) with prolonged isolated thrombocytopenia had a statistically significant lower absolute megakaryocyte count in bone marrow biopsies performed before transplantation and more than 30 days after transplantation compared to control patients (aged 4 months to 50 years, mean 31 years) who underwent HSCT (four sibling HLA-identical, four autologous bone marrow, four autologous peripheral stem cell) for similar conditions. No apparent differences were seen in size of megakaryocytes, nuclear-cytoplasmic ratios, or clustering of megakaryocytes. Overall marrow cellularities were similar in the three groups. These findings suggest that decreased differentiation of megakaryocytes from stem cells, rather than ineffective platelet production or peripheral destruction of platelets, causes prolonged isolated thrombocytopenia in HSCT patients. Low megakaryocyte counts prior to HSCT may be a useful prognostic indicator, as this feature was associated with the development of prolonged isolated thrombocytopenia.


PACAP deficiency sensitizes nigrostriatal dopaminergic neurons to paraquat-induced damage and modulates central and peripheral inflammatory activation in mice.
Exposure to the pesticide paraquat (PQ) increases the risk of Parkinson's disease (PD), and its effect may be modulated by genetic or other environmental factors. The neuropeptide PACAP (pituitary adenylyl cyclase-activating polypeptide, Adcyap1) has been shown to enhance tyrosine hydroxylase (TH) and VMAT2 expression, protect dopaminergic (DA) neurons against the neurotoxin 6-hydroxydopamine, regulate neuronal mitochondria, and inhibit inflammation. Decreased expression of PACAP may thus interact with environmental factors such as PQ to increase the risk of PD. To mimic a low level environmental exposure to PQ, wild type (WT) and PACAP knockout (KO) mice were given a single [10 mg/kg] dose of PQ, a regimen that did not induce the loss of TH expression or DA neurons in WT mice. This treatment selectively reduced the number of TH-positive cell bodies in the substantia nigra pars compacta (SNpc) selectively in PACAP KO mice. Because inflammation is also a risk factor for PD, we performed a quantitative analysis of SNpc Iba⁺ microglia. As expected, PQ increased the number of larger microglial profiles, indicative of activation, in WT mice. Strikingly, microglial activation was already evident in PACAP KO mice in the basal state. PQ caused no further activation in these mice, although tumor necrosis factor-α gene expression was enhanced. In the periphery, PQ had no effects on the abundance of proinflammatory Th1 or Th17 cells in WT mice, but increased the numbers of anti-inflammatory regulatory T cells (Tregs). PACAP KO mice, in contrast, had elevated numbers of Th17 cells after PQ, and the induction of Tregs was impaired. The results indicate that endogenous PACAP acts to maintain the integrity of DA neurons during exposure to PQ, an action that may be linked to its ability to regulate microglia and/or other immune cells.


The fate of mitochondria during platelet activation.
Blood platelets undergo several successive motor-driven reorganizations of the cytoskeleton when they are recruited to an injured part of a vessel. These reorganizations take place during the platelet activation phase, the spreading process on the injured vessel or between fibrin fibers of the forming clot, and during clot retraction. All these steps require a lot of energy, especially the retraction of the clot when platelets develop strong forces similar to those of muscle cells. Platelets can produce energy through glycolysis and mitochondrial respiration. However, although resting platelets have only 5 to 8 individual mitochondria, they produce adenosine triphosphate predominantly via oxidative phosphorylation. Activated, spread platelets show an increase in size compared with resting platelets, and the question arises as to where the few mitochondria are located in these larger platelets. Using expansion microscopy, we show that the number of mitochondria per platelet is increased in spread platelets. Live imaging and focused ion beam-scanning electron microscopy suggest that a mitochondrial fission event takes place during platelet activation. Fission is Drp1 dependent because Drp1-deficient platelets have fused mitochondria. In nucleated cells, mitochondrial fission is associated with a shift to a glycolytic phenotype, and using clot retraction assays, we show that platelets have a more glycolytic energy production during clot retraction and that Drp1-deficient platelets show a defect in clot retraction.


Nanopatterning reconfigurable magnetic landscapes via thermally assisted scanning probe lithography.
The search for novel tools to control magnetism at the nanoscale is crucial for the development of new paradigms in optics, electronics and spintronics. So far, the fabrication of magnetic nanostructures has been achieved mainly through irreversible structural or chemical modifications. Here, we propose a new concept for creating reconfigurable magnetic nanopatterns by crafting, at the nanoscale, the magnetic anisotropy landscape of a ferromagnetic layer exchange-coupled to an antiferromagnetic layer. By performing localized field cooling with the hot tip of a scanning probe microscope, magnetic structures, with arbitrarily oriented magnetization and tunable unidirectional anisotropy, are reversibly patterned without modifying the film chemistry and topography. This opens unforeseen possibilities for the development of novel metamaterials with finely tuned magnetic properties, such as reconfigurable magneto-plasmonic and magnonic crystals. In this context, we experimentally demonstrate spatially controlled spin wave excitation and propagation in magnetic structures patterned with the proposed method.


Additional techniques adopted for major vascular anastomoses during orthotopic liver transplantation.
Of 372 patients who underwent liver transplants between January 1985 and March 1995, 7 required variations in vascular anastomoses due to discrepancies in the size-match of the donor and recipient vessels, the presence of a thrombosed hepatic artery or portal vein, or complete absence of the portal vein. The techniques described herein enabled us to perform successful transplantation in all patients. Although rethrombosis of the portal vein developed in one patient after a third transplant, this patient remains clinically well. The use of an operating microscope and the harvesting of an extended length of donor superior mesenteric vein or vascular grafts of the donor iliac or saphenous vessels, for potentially difficult transplants, are invaluable techniques.


Clinical inquiry: what risk factors contribute to C difficile diarrhea?
Certain antibiotics and using 3 or more antibiotics at one time are associated with Clostridium difficile-associated diarrhea. Hospital risk factors include proximity to other patients with C difficile and longer length of stay. Patient risk factors include advanced age and comorbid conditions. Acid suppression medication is also a risk factor for CDAD.


Genetic characterization of dengue virus serotype 1 circulating in Reunion Island, 2019-2021, and the Seychelles, 2015-2016.
An unprecedent increase in the number of cases and deaths reported from dengue virus (DENV) infection has occurred in the southwestern Indian ocean in recent years. From 2017 to mid-2021 more than 70,000 confirmed dengue cases were reported in Reunion Island, and 1967 cases were recorded in the Seychelles from 2015 to 2016. Both these outbreaks displayed similar trends, with the initial circulation of DENV-2 which was replaced by DENV-1. Here, we aim to determine the origin of the DENV-1 epidemic strains and to explore their genetic characteristics along the uninterrupted circulation, particularly in Reunion. Nucleic acids were extracted from blood samples collected from dengue positive patients; DENV-1 was identified by RT-qPCR. Positive samples were used to infect VERO cells. Genome sequences were obtained from either blood samples or infected-cell supernatants through a combination of both Illumina or MinION technologies. Phylogenetic analyses of partial or whole genome sequences revealed that all DENV-1 sequences from Reunion formed a monophyletic cluster that belonged to genotype I and were closely related to one isolate from Sri Lanka (OL752439.1, 2020). Sequences from the Seychelles belonged to the same major phylogenetic branch of genotype V, but fell into two paraphyletic clusters, with greatest similarity for one cluster to 2016-2017 isolate from Bangladesh, Singapore and China, and for the other cluster to ancestral isolates from Singapore, dating back to 2012. Compared to publicly available DENV-1 genotype I sequences, fifteen non-synonymous mutations were identified in the Reunion strains, including one in the capsid and the others in nonstructural proteins (NS) (three in NS1, two in NS2B, one in NS3, one in NS4B, and seven in NS5). In contrast to what was seen in previous outbreaks, recent DENV-1 outbreaks in Reunion and the Seychelles were caused by distinct genotypes, all likely originating from Asia where dengue is (hyper)endemic in many countries. Epidemic DENV-1 strains from Reunion harbored specific non-synonymous mutations whose biological significance needs to be further investigated.


A mutant of Mycobacterium smegmatis defective in dipeptide transport.
A mutant of Mycobacterium smegmatis unable to use the dipeptide carnosine (beta-alanyl-L-histidine) as a sole carbon or nitrogen source was isolated. Carnosinase activity and the ability to grow on beta-Ala and/or L-His were similar in the mutant and the wild type. However, the mutant showed significant impairment in the uptake of carnosine. This study is the first description of a peptide utilization mutant of a mycobacterium.


Establishment and characterization of adult human gastric epithelial progenitor-like cell lines.
Human gastric epithelial stem/progenitor cells are important for stomach homeostasis; however, the in vitro culture system of these cells remains immature. Although three-dimensional (3D) organoid culture has fundamentally changed the in vitro study of gastrointestinal tract, its use is limited by inaccessible luminal compartment, and difficulties of imaging and manipulation. To overcome these limitations of 3D organoid culture system, we established adult human gastric epithelial progenitor-like (hGEPL) cell lines using a novel robust monolayer cell culture system. We established an in vitro gel-based monolayer culture system for normal human adult gastric epithelium, and compared it with traditional two-dimensional (2D) and 3D organoid culture systems using transcriptomics, immunofluorescence and cell viability experiments. At the same time, we used single-cell transcriptomics to compare the differences of the hGEPL cells in conditioned medium (Cond.) and in chemically defined medium (Chem.), the two most common media for organoid culture, in maintaining the stemness and proliferative activity of hGEPL cells. Finally, we explored the role of key niche factors in inducing hGEPL cell differentiation. The hGEPL cells were similar to the in vivo gastric epithelial stem/progenitor cells, which could stably proliferate in culture for a long time. Based on the established culture system, we explored signalling pathways that were important for the homeostasis of hGEPL cells. We found that after blocking the WNT signalling pathway or activating the BMP signalling pathway, hGEPL cells could differentiate into mucous surface cells. Our culture system of hGEPL cells from adults is robust and easy to operate, and has the transformative potential of personalized and precision medicine, laying a solid foundation for studying the self-renewal and differentiation potentials of gastric epithelial stem/progenitor cells as well as modelling of related gastric diseases.


Evaluation of internal fixation techniques for condylar head fractures: A finite element analysis and comparison.
This study evaluated optimum stability of different screw techniques for condylar head fractures (CHF) (P close to an M fracture with the lateral pole preserved according to AO classification 2014) by finite element analysis (FEA) and provided a biomechanical basis for clinical treatment. Four CHF fixation models were evaluated: (A) single bicortical screw, (B) 2 bicortical screws, (C) 1 bicortical screw and 1 monocortical screw (used as a positional screw) inserted via a 2-hole titanium plate, and (D) 2 bicortical screws inserted via a titanium plate. Stresses were calculated (FEA) to measure mechanical properties. The displacement for A and C was larger than for B and D. The maximum stress on the screws for A and C exceeded their breaking limit but was safe for B and D. The stress on the titanium plate for C and D was safe. The stress on bone for A and C was larger than for B and D. The 2 bicortical screw fixation reduced the stress on implanted materials and surrounding bone tissue. Titanium plates further alleviated the lever action. Two bicortical screw fixation was more reliable for CHF, and early postoperative loading and functional training can be expected.


Pair housing affects anxiety-like behaviors induced by a social but not by a physiological stressor in male Swiss mice.
The role of pair housing in the modulation of anxiety-like behaviour in socially and physiologically stressed mice was investigated. The protocol of psychosocial stress consisted of submitting male adult mice to daily social confrontation with a male conspecific for a period of thirteen days. In an attempt to study a possible effect of pair housing as a social support, each male mouse was housed with a female throughout the period of experimentation, except during the agonistic interactions. As a physiological stressor, 10(9) sheep red blood cells (SRBC)/ml were injected intraperitoneally on the 1st and 7th days of the experiments. The respective control groups were as follows: non-socially stressed, non-pair housed and saline-injected mice. The humoral immune response was analysed by haemagglutination assay. The level of anxiety-like behaviours was measured in the elevated plus-maze test on the 13th day of the experiment. As a result, no significant changes in humoral immunity to SRBC were observed in mice subjected to social confrontation in a neutral arena as compared to non-socially stressed mice. As a consequence, no effect of pair housing on humoral immunity to SRBC could be evaluated. Concerning the effects of pair housing on the anxiety-like behaviours, it was possible to demonstrate that the pair housing proved to be effective in modulating anxiety-like behaviour, although in the stressed groups the percentage of time in the open arms and the time in risk assessment did not change in a symmetrical opposite form, as expected. The physiological stressor induced an anxiety-like behaviour that was not reversed by the pair housed condition. This suggests that different types of stressors activate different neural and peripheral pathways, which may or may not be modulated by pair housing, a finding that deserves our attention as a way to better understand the mechanisms that influence adaptations to stress.


Excellent outcome of living donor liver transplantation in patients with hepatopulmonary syndrome: a single centre experience.
Hepatopulmonary syndrome (HPS) worsens the prognosis of cirrhosis and liver transplantation is only definitive treatment. There is paucity of data about role of living donor liver transplantation (LDLT) in HPS. Fourteen patients with HPS and cirrhosis who underwent LDLT were prospectively included. HPS was defined as PaO2 < 80 mmHg in presence of demonstrable macro-aggregated albumin (MAA) scan shunt fraction >6%. The study group composed of 11 male and three female patients, mean age 50.3 ± 8.6 yr. Most common presentations were dyspnea (92.8%), cyanosis (78.5%) and clubbing (64.2%). Mean model for end-stage liver disease (MELD) score was 18.2 ± 4.7, mean MAA shunt fraction was 23.0 ± 13.2%, mean PaO2 was 58.7 ± 8.4 mmHg. Two patients had very severe HPS (PaO2 <50 mmHg), five had severe HPS (PaO2 >50 <60 mmHg) and seven had moderate HPS (PaO2 >60 <80 mmHg). All patients underwent right lobe LDLT. The overall time to extubation was 2 (1-32 days) and for hospital stay was 20 (17-46 days). The main complications in post-LT course were infection in 57% (cytomegalovirus or bacterial). All the patients are alive and off oxygen at a mean follow up of 29 ± 25 months. We report one of the largest series of LDLT in HPS which has shown excellent results.


Relative prevalence and outcome of fetal posterior fossa abnormality.
To find out the relative incidence and outcome of posterior fossa abnormality (PFA) in terms of survival at birth until 2 years of age. We conducted a prospective study; all fetuses diagnosed with posterior fossa abnormality were followed-up. The outcome was observed with respect to survival, the presence of associated anomalies, the existence of developmental delay after a telephonic interview. Out of 2703 children with congenital anomalies, 921 (34.1%) had a central nervous system defect; 76 cases of PFA were fully followed. Dandy-Walker malformation (DWM) was present in 50% (38/76), mega cisterna magna 18.4% (14/76), Blake pouch cyst 13.2% (10/76), vermian hypoplasia (VH) 13.2% (10/76) and arachnoid cyst 5.2% (4/76). The diagnosis was possible before 20 weeks in only 12 (15.8%) cases. The mean gestational age at delivery was 34.7 ± 6.7 weeks. Associated anomalies were seen in 35/76 (46.1%) cases. A total of 35/76 (46.1%) survived after 2 years; there was developmental delay in 9.2% of cases. There is a large variation in the outcome of PFA depending upon the type of anomaly. Associated anomalies are common in VH and DWM, making their prognosis worse.


[Anterior semifundoplication in the treatment of gastroesophageal reflux disease: long-term results following conventional surgery].
According to randomized studies, semifundoplication in the treatment of gastroesophageal reflux disease (GERD) is related to lower postoperative dysphagia rates than with fundoplication in comparable reflux controls. However there is a lack of long-term results. The object of this study was therefore to determine the influence of partial anterior fundoplication on the long-term clinical outcome (>1 year) in patients with GERD. From December 1986 until May 2000, 100 patients suffering from GERD were operated on. Seven of them were not evaluated, four because of revisional surgery, two because of preceding multiple abdominal operations (MEN, colitis ulcerosa), and one because of Nissen fundoplication. In all, the perioperative results of 93 patients (51 men and 42 women aged 21 to 86 years) were evaluated. Fourteen patients died during the follow-up period and two were lost because of changes in address. Thus, 77 patients with a median follow-up of 88 months (range 15-94) were interviewed with a list of standardized questions concerning reflux control and dysphagia. The median operation time was 110 m (range 55-270). In one patient, an esophageal mucosal tear was detected intraoperatively and promptly repaired. Postoperatively, 71.4% (55/77) had no reflux complaints, 85.7% (66/77) had dysphagia, 66.2% took no further medication, and 31.2% (24/77) continued taking medication. The median interval free of symptoms was 25.5 months, and in 40.9% of the patients, symptoms recurred within the first year after operation. Five patients were in need of reoperation. A total of 77.9% of the patients were satisfied with the results of the operation. With regard to reflux control, long-term results of anterior semifundoplication are comparable to those of fundoplication. However, considering postoperative dysphagia, the technically easier anterior semifundoplication is less eventful and therefore a good alternative which in the long run shows good results after laparoscopic intervention.


Cardiac-specific deletion of SOCS-3 prevents development of left ventricular remodeling after acute myocardial infarction.
The study investigated the role of myocardial suppressor of cytokine signaling-3 (SOCS3), an intrinsic negative feedback regulator of the janus kinase and signal transducer and activator of transcription (JAK-STAT) signaling pathway, in the development of left ventricular (LV) remodeling after acute myocardial infarction (AMI). LV remodeling after AMI results in poor cardiac performance leading to heart failure. Although it has been shown that JAK-STAT-activating cytokines prevent LV remodeling after AMI in animals, little is known about the role of SOCS3 in this process. Cardiac-specific SOCS3 knockout mice (SOCS3-CKO) were generated and subjected to AMI induced by permanent ligation of the left anterior descending coronary artery. Although the initial infarct size after coronary occlusion measured by triphenyltetrazolium chloride staining was comparable between SOCS3-CKO and control mice, the infarct size 14 days after AMI was remarkably inhibited in SOCS3-CKO, indicating that progression of LV remodeling after AMI was prevented in SOCS3-CKO hearts. Prompt and marked up-regulations of multiple JAK-STAT-activating cytokines including leukemia inhibitory factor and granulocyte colony-stimulating factor (G-CSF) were observed within the heart following AMI. Cardiac-specific SOCS3 deletion enhanced multiple cardioprotective signaling pathways including STAT3, AKT, and extracellular signal-regulated kinase (ERK)-1/2, while inhibiting myocardial apoptosis and fibrosis as well as augmenting antioxidant expression. Enhanced activation of cardioprotective signaling pathways by inhibiting myocardial SOCS3 expression prevented LV remodeling after AMI. Our data suggest that myocardial SOCS3 may be a key molecule in the development of LV remodeling after AMI.


Use of an oxygen therapeutic as an adjunct to intraoperative autologous donation to reduce transfusion requirements in patients undergoing coronary artery bypass graft surgery.
The benefits of intraoperative autologous donation (IAD) in reducing the need for allogeneic blood transfusion in surgery have been debated for several years. The purpose of this study was to determine if IAD alone or in conjunction with hemoglobin raffimer (HR) confers a reduction in red cell or blood component transfusion compared with results in standard clinical practice. The Phase III clinical trial was a multicenter, randomized, double-blind study to determine the efficacy and safety of HR versus 10% pentastarch when used to facilitate IAD in 299 patients undergoing primary coronary artery bypass grafting. The patients received HR or pentastarch as an adjunct to IAD immediately before cardiopulmonary bypass. Results were compared with transfusion requirements for 150 matched patients in the reference group. The frequency of allogeneic RBC transfusion in the HR, pentastarch, and reference groups was 56%, 76%, and 95%, respectively. The number of allogeneic red cell units used was 49 in the HR group, 104 in the pentastarch group, and 480 in the reference group (p < 0.001). The total number of non-RBC units administered was 150 in the HR group, 238 in the pentastarch group, and 270 in the reference group. In this study, patients treated with HR in conjunction with IAD received fewer transfusions overall and a lower volume of allogeneic RBCs and non-RBC allogeneic blood products than did the two comparison groups. This confers a real benefit on the overall blood supply by decreasing use and increasing availability.


[Tasks and function of the German Medical Review Board of the Statutory Health Insurance (MDK). Latest aspects in dermatology in view of social medicine].
The German Medical Review Board of the Statutory Health Insurance (MDK) is a sociomedical service for impartial medical opinion and consultation. Together with the statutory health insurance companies it takes part in shaping the public health care system. Many recent developments are changing the german health insurance system. This article illustrates the complex sociomedical framework of MDK's activities in relation to current issues in dermatology. The article focuses on: new diagnostic and therapeutic methods, drugs (off-label-use), out-patient and in-patient dermatological rehabilitation and hospital treatment.


Cancer cell-targeted nanoprobe for multilayer imaging of diverse biomarkers and precise photodynamic therapy.
A novel multifunctional nanoprobe was designed for cancer cell targeted multilayer imaging of two cancer biomarkers. Based on the proposed method, in situ imaging of membrane MUC1 mucin and cytoplasmic microRNA miR-21 coupled with precise photodynamic therapy was achieved.


Upper airways colonisation of Streptococcus pneumoniae in adults aged 60 years and older: A systematic review of prevalence and individual participant data meta-analysis of risk factors.
Colonisation with Streptococcus pneumoniae can lead to invasive pneumococcal disease and pneumonia. Pneumococcal acquisition and prevalence of colonisation are high in children. In older adults, a population susceptible to pneumococcal disease, colonisation prevalence is reported to be lower, but studies are heterogeneous. This is a systematic review and meta-analysis of prevalence of, and risk factors for, pneumococcal colonisation in adults ≥ 60 years of age (PROSPERO #42016036891). We identified peer-reviewed studies reporting the prevalence of S. pneumoniae colonisation using MEDLINE and EMBASE (until April 2016), excluding studies of acute disease. Participant-level data on risk factors were sought from each study. Of 2202 studies screened, 29 were analysable: 18 provided participant-level data (representing 6290 participants). Prevalence of detected pneumococcal colonisation was 0-39% by conventional culture methods and 3-23% by molecular methods. In a multivariate analysis, colonisation was higher in persons from nursing facilities compared with the community (odds ratio (OR) 2•30, 95% CI 1•26-4•21 and OR 7•72, 95% CI 1•15-51•85, respectively), in those who were currently smoking (OR 1•69, 95% CI 1•12-2•53) or those who had regular contact with children (OR 1•93, 95%CI 1•27-2•93). Persons living in urban areas had significantly lower carriage prevalence (OR 0•43, 95%CI 0•27-0•70). Overall prevalence of pneumococcal colonisation in older adults was higher than expected but varied by risk factors. Future studies should further explore risk factors for colonisation, to highlight targets for focussed intervention such as pneumococcal vaccination of high-risk groups. No funding was required.


Stage IB carcinoma of the cervix, the Norwegian Radium Hospital, 1968-1970: results of treatment and major complications. I. Lymphedema.
Different methods of treatment have been debated over the years for Stage IB carcinoma of the cervix, especially in view of the results and complications. We have reviewed the experience of the Norwegian Radium Hospital for carcinoma of the cervix, Stage IB, for the years 1968, 1969, and 1970; most of the patients receive preoperative intrauterine and intravaginal radium, followed by radical hysterectomy and pelvic lymphadenectomy; if the nodes are involved, they also receive external pelvic irradiation. During this period, 437 patients had Stage IB; the 5 year survival was 80.2% (not corrected). We focused our attention on lymphedema following such a method of treatment; of 402 patients operated upon, 23.4% developed lymphedema ranging from mild to moderate to severe; 20 patients (5%) had severe lymphedema; factors involved are analyzed.


Genetic transformation of rare Verbascum eriophorum Godr. plants and metabolic alterations revealed by NMR-based metabolomics.
To develop a protocol to transform Verbascum eriophorum and to study the metabolic differences between mother plants and hairy root culture by applying NMR and processing the datasets with chemometric tools. Verbascum eriophorum is a rare species with restricted distribution, which is poorly studied. Agrobacterium rhizogenes-mediated genetic transformation of V. eriophorum and hairy root culture induction are reported for the first time. To determine metabolic alterations, V. eriophorum mother plants and relevant hairy root culture were subjected to comprehensive metabolomic analyses, using NMR (1D and 2D). Metabolomics data, processed using chemometric tools (and principal component analysis in particular) allowed exploration of V. eriophorum metabolome and have enabled identification of verbascoside (by means of 2D-TOCSY NMR) as the most abundant compound in hairy root culture. Metabolomics data contribute to the elucidation of metabolic alterations after T-DNA transfer to the host V. eriophorum genome and the development of hairy root culture for sustainable bioproduction of high value verbascoside.


Spinal changes associated with mechanical hypersensitivity in a model of Guillain-Barré syndrome.
Guillain-Barré syndrome (GBS) is an inflammatory disease of the peripheral nervous system which can cause pain via mechanisms that are poorly understood. Here, we show that in rat experimental autoimmune neuritis (EAN) mechanical allodynia developed up to 9 days before the onset of detectable neurological deficits. Allodynia was associated with an increase in the number of microglial cells in the dorsal horn of the spinal cord. The expression of the chemokine CX3CL1 (fractalkine) and its receptor CX3CR1 were also higher in EAN than in control dorsal horns suggesting spinal microglia and CX3CL1/CX3CR1 may play a role in the pain-like behaviour.


Prophylactic, therapeutic and neutralizing effects of zinc oxide tetrapod structures against herpes simplex virus type-2 infection.
The attachment of Herpes simplex virus type-2 (HSV-2) to a target cell requires ionic interactions between negatively charged cell surface co-receptor heparan sulfate (HS) and positively charged residues on viral envelop glycoproteins, gB and gC. Effective blocking of this first step of HSV-2 pathogenesis demonstrates significant prophylactic effects against the viral disease; any in vitro therapeutic effects of blocking this interaction, however, are not clear. Here, we provide new evidence that zinc oxide tetrapod micro-nanostructures synthesized by flame transport approach significantly block HSV-2 entry into target cells and, in addition, demonstrate the potential to stop the spread of the virus among already infected cells. The zinc oxide tetrapods (ZnOTs) also exhibit the ability to neutralize HSV-2 virions. Natural target cells such as human vaginal epithelial and HeLa cells showed highly reduced infectivity when infected with HSV-2 virions that were pre-incubated with the ZnOTs. The mechanism behind the ability of ZnOTs to prevent, neutralize or reduce HSV-2 infection relies on their ability to bind the HSV-2 virions. We used fluorescently labeled ZnOTs and GFP-expressing HSV-2 virions to demonstrate the binding of the ZnOTs with HSV-2. We also show that the binding and hence, the antiviral effects of ZnOTs can be enhanced by illuminating the ZnOTs with UV light. Our results provide new insights into the anti-HSV-2 effects of ZnOT and rationalize their development as a HSV-2 trapping agent for the prevention and/or treatment of infection. The observed results also demonstrate that blocking HSV-2 attachment can have prophylactic as well as therapeutic applications.


A preliminary study investigating class characteristics in the Gurmukhi handwriting of 1st and 2nd generation Punjabis.
Gurmukhi is a written script of the Punjabi language spoken by 104 million people worldwide. It has been previously shown in a study of Punjabi residents to contain several unique class characteristics. In this paper these class characteristics and others were analysed in both 1st generation and 2nd generation Punjabi decedents residing in the United Kingdom. Using the Pearson Chi-squared test, eight characteristic features were found to be statistically different in the Gurmukhi handwriting of the two populations (p > 0.01). Additionally there are several changes in previously identified class characteristics, such as script type and angularity of characters, between the 1st generation and 2nd generation Punjabi populations. These class characteristics may be of value to forensic document examiners and allow them to identify the population and the generation of the writer of a suspect document.


Clinical study of brain retraction in different approaches and diseases.
Pressure of brain retraction was measured with a strain gauge spatula in 31 cases of 21 tumours and 10 aneurysms. They were operated on via different approaches: subfrontal (10 cases), interhemispheric (5), subtemporal (7) and suboccipital (9). The patients' age ranged from 11 to 74 years (average 50). The recorded data were averaged for every 5 minutes and divided into two groups: the first was data taken in the initial stage during the approach to the lesions, and the second were those taken during procedures at and around the lesion. Significant difference in the pressure among different approaches was found in the tumour group approached by the subtemporal route compared with other routes (p less than 0.005). The retraction pressures in the tumour cases were higher during the approach (31 +/- 15.5 torr) than during the main procedure (12 +/- 9.2 torr) (p less than 0.025). On the other hand, the pressures in the aneurysm cases were the reverse, being lower during the approach (22 +/- 15.2 torr) than during the main procedure (38 +/- 17.7 torr) (p less than 0.01). Early-stage operations for aneurysm required a higher retraction pressure than delayed operations.


Magnitude and causes of first-line antiretroviral therapy regimen changes among HIV patients in Ethiopia: a systematic review and meta-analysis.
Antiretroviral therapy (ART) has markedly decreased the morbidity and mortality due to HIV/AIDS. ART regimen change is a major challenge for the sustainability of human immunodeficiency virus (HIV) treatment program. This is found to be a major concern among HIV/AIDS patients in a resource-limited setting, where treatment options are limited. The aim of this review is to generate the best available evidence regarding the magnitude of first-line antiretroviral therapy regimen change and the causes for regimen change among HIV patients on ART in Ethiopia. The reviewed studies were accessed through electronic web-based search strategy from PubMed Medline, EMBASE, Hinari, Springer link and Google Scholar. Data were extracted using Microsoft Excel and exported to Stata software version 13 for analyses. The overall pooled estimation of outcomes was calculated using a random-effect model of DerSimonian-Laird method at 95% confidence level. Heterogeneity of studies was determined using I2 statistics. For the magnitude of regimen change, the presence of publication bias was evaluated using the Begg's and Egger's tests. The protocol of this systematic review and meta-analysis was registered in the Prospero database with reference number ID: CRD42018099742. The published methodology is available from: https://www.crd.york.ac.uk/PROSPERO/display_record.php?RecordID=99742 . A total of 22 studies published between the years 2012 and 2018 were included. Out of 22 articles, 14 articles reported the magnitude of regimen change and consisted of 13,668 HIV patients. The estimated national pooled magnitude of regimen change was 37% (95% CI: 34, 44%; Range: 15.1-63.8%) with degree of heterogeneity (I2), 98.7%; p-value < 0.001. Seventeen articles were used to identify the causes for first-line antiretroviral therapy regimen change. The major causes identified were toxicity, 58% (95% CI: 46, 69%; Range: 14.4-88.5%); TB co-morbidity, 12% (95% CI: 8, 16%; Range: 0.8-31.7%); treatment failure, 7% (95% CI: 5, 9%; Range: 0.4-24.4%); and pregnancy, 5% (95% CI: 4, 7%; Range: 0.6-11.9%). The original first-line regimen was changed in one-third of HIV patients on ART in Ethiopia. Toxicity of the drugs, TB co-morbidity, treatment failure, and pregnancy were the main causes for the change of the first-line regimen among HIV patients on antiretroviral therapy.


Vibrio vulnificus IlpA induces MAPK-mediated cytokine production via TLR1/2 activation in THP-1 cells, a human monocytic cell line.
Vibrio vulnificus is a pathogenic bacterium causing primary septicemia, which is followed by a classical septic shock pathway including an overwhelming inflammatory cytokine response. V. vulnificus IlpA is a potent immunogenic lipoprotein that triggers cytokine production in human monocytes by activating the toll-like receptor 2 (TLR2). In this study, we further defined the IlpA signaling pathways involved in cytokine production in the human monocytic cell line, THP-1. TLR2 was involved in cytokine production by complexing with TLR1, but not with TLR6. MyD88 was necessary for IlpA-induced cytokine expression through TLR1/TLR2. Three mitogen activated protein kinases (MAPK), p38, ERK1/2, and JNK, were activated in THP-1 cells stimulated with recombinant IlpA (rIlpA). Selective inhibition of each MAPK resulted in significant decrease of rIlpA-induced cytokine production. Especially, functional TLR2 was necessary for IlpA-induced activation of p38 and JNK. IlpA augmented the DNA-binding activity of nuclear factor-kappaB (NF-κB) and activator protein-1 (AP-1) transcriptional factors to their recognition sites in THP-1 cells. These results suggest that serial activation of TLR1/TLR2, MyD88, the three MAPKs, and NF-κB/AP-1 comprises the signaling pathway responsible for proinflammatory cytokine production by V. vulnificus IlpA.


Molecular separation in the lipid bilayer medium: electrophoretic and self-spreading approaches.
Molecular separation in a microchannel is a key technology for future miniature devices. Because of growing advances in microfabrication techniques, we now have various choices of microscopic molecular separation systems. Recent progress in this field is reviewed in this manuscript. In particular, the use of the lipid bilayer as a separation medium is highlighted, because of its possible application to the manipulation of relatively small biomaterials such as membrane proteins and lipids. In this context, an approach based on electrophoresis is reviewed for molecular separation in the bilayer. Although the methods based on electrophoresis are effective, we will also focus on their limitation, i.e., only charged molecules can be manipulated. To solve this dilemma, we review new techniques based on the self-spreading nature of the lipid bilayer. In this method, there is no need to input an external field, such as an electric field, to achieve molecular separation. Phenomenological insights into the self-spreading nature and newly proposed molecular separation effects are shown in detail. This novel concept enables us to establish a completely unbiased molecular separation system in future microscopic and nanoscopic devices.


The Pseudogene BMEA_B0173 Deficiency in Brucella melitensis Contributes to M-epitope Formation and Potentiates Virulence in a Mice Infection Model.
It is widely acknowledged that pseudogenes play important roles in bacterial diversification and evolution and participate in gene regulation and RNA interference (RNAi). However, the function of most pseudogenes in Brucella spp remains poorly understood, warranting further studies.To comprehensively analyze the function of the pseudogenes BMEA_B0173 in Brucella melitensis strain 63/9, a BMEA_B0173 in-frame deleted mutant strain was constructed. Then, the phenotypes of the mutant strain, such as growth characteristics and bacterial virulence, were assessed in mice infection models. Finally, iTRAQ analysis was performed to investigate the gene expression profile affected by the pseudogenes BMEA_B0173. In this study, we found that BMEA_B0173 deletion exhibited increased agglutination with M monospecific sera. In a mouse model of chronic infection, the BMEA_B0173 deletion strain displayed increased colonization in the spleen compared to the wild-type pathogen. The iTRAQ assay revealed that 252 proteins were differentially expressed between the BMEA_B0173 deletion and the wild-type strains. In addition, deletion of BMEA_B0173 significantly increased the expression of proteins involved in the denitrification pathway, iron metabolism, and several transcriptional regulators, which might cause increased virulence of the mutant strain. In conclusion, this study preliminary uncovered the function of the pseudogene BMEA_B0173 in Brucella melitensis 63/9 and provided novel insights for studying the pathogenesis of Brucella strains.


The perception of translucency from surface gloss.
Translucent objects (like fruit and wax) reflect and transmit incident light to generate complex retinal image structure. Understanding how we visually perceive translucency from these images is challenging, but previous studies have demonstrated that perceived shape and shading is important for perceiving translucency. We considered the possibility that perceived translucency might also depend on 3D shape inferred from surface gloss (i.e., shape from specular highlights). Here, we performed experiments to test whether interactions between specular and non-specular image properties generated by different 3D shape information influences perceived translucency. Results revealed that perceived translucency could be explained by incongruence in 3D shape used to generate specular and non-specular image components. We proposed a new computational model based on measurable image features informative of shading relative to specular highlights that accounted for 59% of the variability in judgments of perceived translucency from the result of 10-fold cross validation. This model was found to outperform other models based on explicit subjective measures of perceived surface shape, suggesting it implicitly taps much of the relevant geometric information necessary for predicting observer judgments of translucency for glossy materials. These results provide new insight into how the visual system might infer translucency from the structure of specular and non-specular shading generated by glossy semi-opaque materials.


Developmental exposure of fetal ovaries and fetal germ cells to endometriosis in an endometriosis model causes differential gene expression in the preimplantation embryos of the first-generation and second-generation embryos.
To characterize multigenerational gene expression anomalies in eight-cell stage embryos associated with developmental exposure to endometriosis. Using an endometriosis model in rats (F0 founder generation) to evaluate gene expression in F1 (fetal exposure) and F2 (fetal germ cell exposure) generation eight-cell stage embryos. Laboratory. Endometriosis model in rats (Endo) and controls (Sham). F0 Endo and Sham rats were bred; half the pregnant rats were killed on gestational day 3 to collect F1 eight-cell stage embryos and the others gestated to term (F1 females). Adult F1 females bred; F2 eight-cell embryos collected. Maintenance of differential gene expression in F1 and F2 generation eight-cell embryos in endometriosis. Developmental exposure to endometriosis altered the gene signaling pathways, with changes found in apoptosis, the cell cycle process, the response to oxidative stress, negative regulation of molecular function, and RNA processing. The apoptotic genes Diablo, Casp3, Parp1, Cad, and Dnaja3 were increased and the Nfkbia transcripts were decreased in F1 Endo versus F1 Sham embryos. In F2 Endo versus Sham embryos, Casp3 and Cad were statistically significantly increased, and Parp1 and Nfkbia tended to be elevated. Fetal and germ cell exposure to endometriosis alters apoptotic gene expression in first- and second-generation eight-cell stage embryos, supporting the hypothesis of multigenerational inheritance resulting from exposure to endometriosis in utero.


[The effect of respiratory exercise on the Hirtz index and thoracic value in subjects with idiopathic scoliosis].
The effects of prolonged gymnastics on respiratory capacity (assessed by means of the Hirtz index and the chest value) were evaluated in patients aged 10-15 yr with scoliotic curves of not more than 30 degrees. The mean overall Hirtz value, initially well below normal, displayed an average improvement of 0.44 (higher in lumbar forms, and decreasing in dorsolumbar, double and dorsal scoliosis). As expected, the chest value was virtually unchanged.


Anti-inflammatory activity of Echinosophora koreensis nakai root extract in lipopolysaccharides-stimulated RAW 264.7 cells and carrageenan-induced mouse paw edema model.
Echinosophora koreensis Nakai is an endemic plant species distributed in a limited area within the Korean province of Gangwon, including the Yanggu-gun, Inje-gun, Cheorwon-gun, Chuncheon-si, and Hongcheon-gun counties. It is used in traditional medicine to treat various disorders, such as fever, skin diseases, diuresis, and neuralgia. This study demonstrated the effects of E. koreensis Nakai root extract (EKRE) on lipopolysaccharide (LPS)-induced inflammatory responses in vitro and in vivo. Cell viability was assessed through a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. Nitric oxide (NO) production was measured using Griess reagent. Interleukin (IL)-6 and tumor necrosis factor (TNF) levels were assessed using enzyme-linked immunosorbent assays. Inducible nitric oxide synthase (iNOS), nuclear factor kappa-B (NF-κB), and mitogen-activated protein kinase (MAPK) expression were assessed using Western blot analysis. To examine the effects of EKRE in vivo, it was administered orally at doses of 50 or 200 mg/kg for 3 days in mice. Edema in the paws was induced through λ-carrageenan injection and measured hourly for up to 5 h using calipers. EKRE markedly suppressed LPS-generated NO, IL-6, and iNOS production in RAW 264.7 cells. Moreover, it suppressed the activation of the NF-κB and MAPK in LPS-stimulated cells. Furthermore, EKRE significantly inhibited carrageenan-induced edema in mouse paws. There were no significant differences in IL-6 and TNF production in paw tissue harvested from mice, but levels decreased at high EKRE concentrations (200 mg/kg). The results of this study provided validation for EKRE-induced inhibition of inflammatory responses in vitro and in vivo. This research suggested that EKRE is a promising treatment for inflammatory disorders.


Pharmacokinetics, safety, and tolerability of single- and multiple-dose exenatide once weekly in Chinese patients with type 2 diabetes mellitus.
This open-label, single-period study assessed the pharmacokinetics, safety, tolerability, and pharmacodynamics of exenatide once weekly (q.w.), following single and multiple weekly subcutaneous (s.c.) injections in native Chinese patients with type 2 diabetes (T2D). Patients (n = 25; mean [±SD] age 51.3 ± 8.2 years; body mass index 25.6 ± 2.4 kg/m(2) ; HbA1c 7.4 ± 1.2%; duration of diabetes 3.1 ± 3.1 years) previously treated with diet modification and exercise alone or incombination with stable metformin doses were enrolled in the study. Twenty-five patients received weekly doses of 2 mg, s.c., exenatide q.w. for 10 weeks, followed by 10 weeks observation. Pharmacokinetic parameters of exenatide, fasting plasma glucose (FPG), HbA1c, and body weight were summarized using descriptive statistics. Steady state plasma exenatide concentrations (299 pg/mL) were attained within 8 weeks. Exenatide q.w. was generally well tolerated, and the majority of adverse events reported were mild in severity. The most frequent study drug-related adverse events were diarrhea and vomiting. Decreases were observed from baseline to 10 weeks in FPG (~3.0 mmol/L), HbA1c (~1.0%), and body weight (~3.8 kg). This is the first clinical trial of exenatide q.w. in native Chinese patients with T2D. The results suggest that exenatide q.w. has a pharmacokinetic profile in this patient population similar to that observed in other ethnic and racial populations, and appears to be safe and generally well tolerated, with the potential to improve glycemic control and decrease body weight without increasing the risk of hypoglycemia.


Sarcoidosis presenting as bilateral vocal cord paralysis from bilateral compression of the recurrent laryngeal nerves from thoracic adenopathy.
Bilateral true vocal fold paralysis is rarely attributable to inflammatory diseases. We describe what appears to be the first case in the medical literature of sarcoidosis presenting as isolated, bilateral true vocal cord paralysis resulting from compressive bilateral mediastinal adenopathy. The presenting symptoms, clinical outcome, radiographs and laryngeal findings are discussed in detail. Sarcoidosis should therefore be added to the differential diagnosis of bilateral vocal fold paralysis.