Interannual variability in the skate assemblage on the South Patagonian shelf and slope.
Observer data from the commercial fishery on the Patagonian shelf and slope around the Falkland Islands (home to an assemblage of >16 skate species (Rajiformes), for which commercial catches have been recorded since 1987), as well as survey data from an area closed to skate target fishing after exploitation, were summarized by species to examine changes in the population status of individual skate species. Total skate catch per unit effort increased significantly in the target fishery since 1994, and four species have made up >85% of all skate catch. Bathyraja brachyurops and Zearaja chilensis increased significantly in catch proportions and abundance from 1994 to 2013. Bathyraja albomaculata and Bathyraja griseocauda decreased significantly before rebounding with trends of increasing abundance. Concurrently, B. brachyurops and Z. chilensis showed decreasing trends in size at 50% maturity in areas where skates continue to be targeted commercially. The increasing abundances and concomitant reductions in size at maturity of B. brachyurops and Z. chilensis suggest either plasticity in life-history traits or a density-dependent growth response to fishing pressure. Bathyraja griseocauda decreased in size at 50% maturity in the area that was closed to skate target fishing, where it was initially larger, but only decreased to the same average size as in the commercially targeted areas. Bathyraja albomaculata and Z. chilensis are IUCN-listed as vulnerable and B. griseocauda is listed as endangered, but their abundance trends since 1994 indicate that these populations are not declining in Falkland waters.


Neuroendocrine predictors of vasoplegia after cardiopulmonary bypass.
Vasoplegia often complicates on-pump cardiac surgery. Systemic inflammatory response induced by extracorporeal circulation represents the major determinant, but adrenal insufficiency and postoperative vasopressin deficiency may have a role. Pathophysiological meaning of perioperative changes in endocrine markers of hydro-electrolyte balance has not still fully elucidated. Objectives of the present research study were to estimate the incidence of vasoplegia in a homogeneous cohort of not severe cardiopathic patients, to define the role of presurgical adrenal insufficiency, to evaluate copeptin and NT-proBNP trends in the perioperative. We conducted a prospective cohort study in the cardiac intensive care unit of a tertiary referral center. We evaluated 350 consecutive patients scheduled for cardiac surgery; 55 subjects completed the study. Both standard and low-dose corticotropin stimulation tests were performed in the preoperative; copeptin and NT-proBNP were evaluated in the preoperative (T0), on day 1 (T1) and day 7 (T2) after surgery. Nine subjects (16.3%) developed vasoplegic syndrome with longer bypass and clamping time (p < 0.001). Reduced response to low-dose ACTH test was not associated to vasoplegia. Preoperative copeptin > 16.9 pmol/L accurately predicted the syndrome (AUC 0.86, 95% CI 0.73-0.94; OR 1.17, 95% CI 1.04-1.32). An evident correlation was observed at 7 days postoperative between NT-proBNP and copeptin (r 0.88, 95% CI 0.8-0.93; p < 0.001). Preoperative impaired response to low-dose ACTH stimulation test is not a risk factor for post-cardiotomic vasoplegia; conversely, higher preoperative copeptin predicts the complication. On-pump cardiac surgery could be an interesting model of rapid heart failure progression.


Single-passage extraction and permeability estimation of sodium in normal dogs lungs.
Bolus injections of 24NaCl, 131I-albumin, and indocyanine green dye were made into the right atria of anesthetized, ventilated dogs. Blood was sampled from the femoral artery, and from the dilution curves, instantaneous extractions, E2(t), and area averaged extractions, E3(t), were calculated for sodium at various plasma flows (Fp). Flow reduction was produced by transient inferior vena cava obstruction. E2(t) and E3(t) within any dilution curve generally started off with a high value, then decreased with time. The E3 that occurred at the peak of the albumin-dilution curve were about 0.11 for plasma flow of 0.75 liter/min and tended to decrease as flow increased. Parallel study of the sodium extravascular space at equilibrium gave values of 0.3-0.4 g of plasma sodium per g of tissue, suggesting that this volume is not infinitely small. Since the E was low it is unlikely that the high initial E(t) and the decreasing E(t) were due to early back flux. Calculation of capillary permeability surface area product [PS = Fp loge (1 - E)] showed an increasing PS with plasma flows. The injection of 9-mum microspheres at low and high total blood flow gave evidence supporting a decrease of capillary surface area (S) with decreasing total blood flow. Regional pulmonary blood flow also showed marked heterogeneity. Because of the low average extraction of sodium in the lung, insensitivity of the method in normal lungs cannot be excluded.


Emergency-aid for self-employed in the Covid-19 pandemic: A flash in the pan?
The self-employed faced strong income losses during the Covid-19 pandemic. Many governments introduced programs to financially support the self-employed during the pandemic, including Germany. The German Ministry for Economic Affairs announced a €50bn emergency-aid program in March 2020, offering one-off lump-sum payments of up to €15,000 to those facing substantial revenue declines. By reassuring the self-employed that the government 'would not let them down' during the crisis, the program had also the important aim of motivating the self-employed to get through the crisis. We investigate whether the program affected the confidence of the self-employed to survive the crisis using real-time online-survey data comprising more than 20,000 observations. We employ propensity score matching, making use of a rich set of variables that influence the subjective survival probability as main outcome measure. We observe that this program had significant effects, with the subjective survival probability of the self-employed being moderately increased. We reveal important effect heterogeneities with respect to education, industries, and speed of payment. Notably, positive effects only occur among those self-employed whose application was processed quickly. This suggests stress-induced waiting costs due to the uncertainty associated with the administrative processing and the overall pandemic situation. Our findings have policy implications for the design of support programs, while also contributing to the literature on the instruments and effects of entrepreneurship policy interventions in crisis situations.


Teleological and referential understanding of action in infancy.
There are two fundamentally different ways to attribute intentional mental states to others upon observing their actions. Actions can be interpreted as goal-directed, which warrants ascribing intentions, desires and beliefs appropriate to the observed actions, to the agents. Recent studies suggest that young infants also tend to interpret certain actions in terms of goals, and their reasoning about these actions is based on a sophisticated teleological representation. Several theorists proposed that infants rely on motion cues, such as self-initiated movement, in selecting goal-directed agents. Our experiments revealed that, although infants are more likely to attribute goals to self-propelled than to non-self-propelled agents, they do not need direct evidence about the source of motion for interpreting actions in teleological terms. The second mode of action-based mental state attribution interprets actions as referential, and allows ascription of attentional states, referential intents, communicative messages, etc., to the agents. Young infants also display evidence of interpreting actions in referential terms (for example, when following others' gaze or pointing gesture) and are very sensitive to the communicative situations in which these actions occur. For example, young infants prefer faces with eye-contact and objects that react to them contingently, and these are the very situations that later elicit gaze following. Whether or not these early abilities amount to a 'theory of mind' is a matter of debate among infant researchers. Nevertheless, they represent skills that are vital for understanding social agents and engaging in social interactions.


Collaborative study of speaker identiication by the voiceprint method.
In the voiceprint method known and unknown voices are compared to determine if they are the same speaker. An unknown voice is recorded over the telephone onto a quality recorder. The known voice is similarly recorded, repeating the questioned message verbatim. An examiner determines, by means of acoustic spectrography and aural analysis, the similarities or differences existing between the known and unknown speakers. A positive conclusion is based on at least 10 pairs of like sounds. Five decisions are available for the examiner to make: positive identification, positive elimination, probable identification, probable elimination, and unable to make a decision with the sample submitted. Seven collaborators, chosen from various parts of the United States, were given recordings of 4 unknown speakers, 2 male and 2 female, to compare with similar recordings of 6 known speakers, 3 male and 3 female. A total of 21 positive identifications and 63 positive eliminations was possible. Twenty correct identifications and 47 correct positive eliminations were made. In 16 tasks, all involving the same unknown sample, collaborators reported they could not make a determination due to the poor quality of the sample. No false identifications were made; however, a trainee examiner, with less than 2 years experience, eliminated a known speaker when in fact a match did exist, thereby falsely eliminating the speaker. Examiners with more than 2 years of experience correctly identified all existing matches. This study indicates that a trained examiner can make very reliable decisions, using the aural and visual methods of comparing known and unknown voices. The voiceprint method has been adopted as official first action.


Borrelia burgdorferi population dynamics and prototype gene expression during infection of immunocompetent and immunodeficient mice.
The population dynamics of Borrelia burgdorferi were quantified by real-time PCR targeting the flaB gene in skin (inoculation site, noninoculation site, and ear), heart (heart base and ventricle), quadriceps muscle, and the tibiotarsal joint at 1, 2, 4, 6, and 8 weeks after intradermal inoculation in C3H and C3H-scid mice. In addition, RNA transcription was assessed for several prototype genes, including flaB, ospA, ospC, dbpA, arp, vlsE, fbp, oppA-2, and p37-42. Spirochete numbers were equivalent in C3H and C3H-scid mice at 1 or 2 weeks and then declined in C3H mice, but they continued to rise and then plateaued in C3H-scid mice. Gene transcription was likewise higher in C3H-scid mice than in C3H mice, particularly at 4 or more weeks of infection. Gene transcription showed variation among tissues, with the highest levels of transcription in heart and joint tissue, which are sites of inflammation.


Hydrogenation of the exocyclic olefinic bond at C-16/C-17 position of ent-kaurane diterpene glycosides of Stevia rebaudiana using various catalysts.
Catalytic hydrogenation of the exocyclic double bond present between C16 and C17 carbons of the four ent-kaurane diterpene glycosides namely rebaudioside A, rebaudioside B, rebaudioside C, and rebaudioside D isolated from Stevia rebaudiana has been carried out using Pt/C, Pd(OH)2, Rh/C, Raney Ni, PtO2, and 5% Pd/BaCO3 to their corresponding dihydro derivatives with 17α and 17β methyl group isomers. Reactions were performed using the above-mentioned catalysts with the solvents methanol, water, and ethanol/water (8:2) under various conditions. Synthesis of reduced steviol glycosides was performed using straightforward chemistry and their structures were characterized on the basis of 1D and 2D NMR spectral data, including a comparison with reported spectral data.


Axons are injured by antigen-specific CD8(+) T cells through a MHC class I- and granzyme B-dependent mechanism.
Axon injury is a central determinant of irreversible neurological deficit and disease progression in patients with multiple sclerosis (MS). CD8(+) lymphocytes (CTLs) within inflammatory demyelinated MS lesions correlate with acute axon injury and neurological deficits. The mechanisms of these correlations are unknown. We interrogated CTL-mediated axon injury using the transgenic OT-I antigen-specific CTL model system in conjunction with a chambered cortical neuron culture platform that permitted the isolated manipulation of axons independent of neuron cell bodies and glia. Interferon gamma upregulated, through a dose dependent mechanism, the axonal expression of functional major histocompatibility complex class I (MHC I) molecules competent to present immunologically-relevant antigens derived from endogenously expressed proteins. Antigen-specific CTLs formed cytotoxic immune synapses with and directly injured axons expressing antigen-loaded MHC I molecules. CTL-mediated axon injury was mechanistically dependent upon axonal MHC I antigen presentation, T cell receptor specificity and axoplasmic granzyme B activity. Despite extensive distal CTL-mediated axon injury, acute neuron cell body apoptosis was not observed. These findings present a novel model of immune-mediated axon injury and offer anti-axonal CTLs and granzyme B as targets for the therapeutic protection of axons and prevention of neurological deficits in MS patients.


Sleep disturbances in depressed and nondepressed pregnant women.
Sleep disturbances and symptoms of depression are common during pregnancy. Both are independent and interrelated risk factors for adverse outcomes. It is unclear the degree to which sleep differs between depressed and nondepressed pregnant women. We sought to (1) describe and compare sleep disturbances in depressed pregnant and nondepressed pregnant women, (2) determine the impact of selective serotonin reuptake inhibitors (SSRI) treatment on sleep, and (3) evaluate whether sleep at 20 weeks is associated with increased depressive symptoms and major depressive disorder (MDD) in later pregnancy. Pregnant women (N = 240) were recruited in the second trimester (20 weeks gestation) and assigned to depressed (N = 59) and nondepressed (N = 181) groups based on a Structured Clinical Interview for DSM-IV diagnosis of major depressive disorder. The Structured Interview Guide for the Hamilton Rating Scale with Atypical Depression Supplement was administered at 20, 30, and 36 weeks gestation from which the sleep variables were obtained. Depressed women had more fragmented sleep at each assessment (P values≤.05). However, the frequency of insomnia symptoms was greater for depressed women only at 20 weeks gestation. SSRI use, regardless of MDD status, did significantly affect several sleep variables. Among the nondepressed women, those with short or longer sleep duration, symptoms of insomnia and long periods of nocturnal waketime had higher Structured Interview Guide for the Hamilton Rating Scale with Atypical Depression Supplement scores later in pregnancy (P values≤.05). At 20 and 30 weeks gestation sleep was more disturbed in depressed pregnant women compared to nondepressed pregnant women. At 36 weeks, sleep was disturbed regardless of depression status or SSRI use. Among the nondepressed women, disturbed sleep in conjunction with SSRI use was associated with higher depressive symptoms.


Survivors of acute myocardial infarction: who is at risk for sudden cardiac death?
Coronary artery disease is the leading cause of death in the United States. Approximately half of the deaths attributable to coronary artery disease are sudden cardiac deaths. A logical approach to prevention of sudden death is to identify those who are at risk and then to initiate effective therapy. Left ventricular dysfunction, frequent ventricular ectopic activity, nonsustained ventricular tachycardia, and late potentials have been identified as markers for increased risk of sudden cardiac death. The sensitivity and specificity of these risk factors vary, and the positive predictive power is less than satisfactory. The value of invasive electrophysiologic testing for risk stratification in the general postinfarction patient population remains unclear. In addition to these diagnostic difficulties, prevention of sudden death also has been limited by imperfect efficacy and potential lethal effects of the currently available antiarrhythmic agents. Automatic implantable defibrillators are effective for aborting sudden death; however, the potential for more general use of automatic defibrillators in asymptomatic but high-risk postinfarction patients has not been evaluated.


[Cultivation of Neisseria meningitidis serogroups A, B and C in a synthetic nutrient broth].
The processes of the cultivation of N. meningitidis, serogroups A, B and C, in a liquid synthetic culture medium have been studied. Strictly group-specific biomass has been obtained. The maximum productivity at all stages of the batch cultivation of N. meningitidis strains 125 and 133 in this medium does not differ from that at similar stages of cultivation in modified Cohen-Wheller semisynthetic medium. In the serotype antigen preparations obtained from N. meningitidis strain 125 grown in the above-mentioned liquid synthetic culture medium basic polypeptides with a molecular weight of 33000, 36000 and 41000 D have been detected. Their presence in N. meningitidis cells is linked with the growth phase of the population.


Prospectively assessed clinical outcomes in concussive blast vs nonblast traumatic brain injury among evacuated US military personnel.
Blast injury has been identified as the signature injury in the conflicts in Iraq and Afghanistan. However it remains to be determined whether fundamental differences may exist between blast-related traumatic brain injury (TBI) and TBI due to other mechanisms. To determine similarities and differences between clinical outcomes in US military personnel with blast-related vs. non-blast-related concussive TBI and to identify the specific domains of impairment that best correlate with overall disability. Prospective cohort study involving active duty US Military personnel evacuated from Iraq or Afghanistan to Landstuhl Regional Medical Center, in Landstuhl, Germany. Four groups of participants were enrolled from 2010 to 2013: (1) blast plus impact complex TBI (n=53), (2) non-blast related TBI with injury due to other mechanisms (n=29), (3) blast-exposed controls evacuated for other medical reasons (n=27) (4) non-blast-exposed controls evacuated for other medical reasons (n=69). All patients with TBI met Department of Defense criteria for concussive (mild) TBI. The study participants were evaluated 6-12 months after injury at Washington University in St Louis. In total, 255 subjects were enrolled in the study, and 183 participated in follow-up evaluations, 5 of whom were disqualified. In-person clinical examinations included evaluation for overall disability, a standardized neurological exam, headache questionnaires, neuropsychological test battery, combat exposure and alcohol use surveys, and structured interview evaluations for post-traumatic stress disorder (PTSD) and depression. Global outcomes, headache severity, neuropsychological performance, and surprisingly even PTSD severity and depression were indistinguishable between the two TBI groups, independent of mechanism of injury. Both TBI groups had higher rates of moderate to severe overall disability than the respective control groups: 41/53 (77%) of blast plus impact TBI and 23/29 (79%) of nonblast TBI vs. 16/27 (59%) of blast-exposed controls and 28/69 (41%) of non-blast-exposed controls. In addition, blast-exposed controls had worse headaches and more severe PTSD than non-blast-exposed controls. Self-reported combat exposure intensity was higher in the blast plus impact TBI group than in nonblast TBI group and was higher in blast-exposed controls than in non-blast-exposed controls. However, combat exposure intensity did not correlate with PTSD severity in the TBI groups, but a modest positive correlation was observed in the controls. Overall outcomes were most strongly correlated with depression, headache severity, and number of abnormalities on neuropsychological testing. However a substantial fraction of the variance in overall outcome was not explained by any of the assessed measures. One potential interpretation of these results is that TBI itself, independent of injury mechanism and combat exposure intensity, is a primary driver of adverse outcomes. Many other important factors may be as yet unmeasured, and adverse outcomes following war-time injuries are difficult to fully explain. clinicaltrials.gov Identifier: NCT01313130.


Urinary Neuropilin-1: A Predictive Biomarker for Renal Outcome in Lupus Nephritis.
At present, Lupus Nephritis (LN) is still awaiting a biomarker to better monitor disease activity, guide clinical treatment, and predict a patient's long-term outcome. In the last decade, novel biomarkers have been identified to monitor the disease, but none have been incorporated into clinical practice. The transmembrane receptor neuropilin-1 (NRP-1) is highly expressed by mesangial cells and its genetic deletion results in proteinuric disease and glomerulosclerosis. NRP-1 is increased in kidney biopsies of LN. In this work we were interested in determining whether urinary NRP-1 levels could be a biomarker of clinical response in LN. Our results show that patients with active LN have increased levels of urinary NRP-1. When patients were divided according to clinical response, responders displayed higher urinary and tissue NRP-1 levels at the time of renal biopsy. Areas under the receiver operating characteristic curve, comparing baseline creatinine, proteinuria, urinary NRP-1, and VEGFA protein levels, showed NRP-1 to be an independent predictor for clinical response. In addition, in vitro studies suggest that NRP-1could promote renal recovery through endothelial proliferation and migration, mesangial migration and local T cell cytotoxicity. Based on these results, NRP-1 may be used as an early prognostic biomarker in LN.


Polarity of recombination in transformation of Streptococcus pneumoniae.
In transformation of Streptococcus pneumoniae DNA enters the cell as single-strand fragments and integrates into the chromosome by homologous recombination. Deletions and insertions of a few hundred base pairs frequently stop the recombination process of a donor strand. In this work we took advantage of such interruptions of recombination to compare the transformation efficiencies of the segments 5'- and 3'-ward from a deletion. The deletion was created in the center of a fragment of the ami locus, and sites around the deletion were labeled by a frameshift generating a restriction site. Heteroduplexes were constructed containing two restriction sites on one strand and two different ones on the complementary strand. ami+ bacteria were transformed with such heteroduplexes. ami- transformants were isolated and individually underwent amplification of the transformed ami region. We have obtained two kinds of amplification products: short when the deletion was integrated, long when recombination stops at the deletion. Each long fragment was tested by the four restriction enzymes to detect which strand and which side of the deletion had recombined. We found that 80% of the cuts were located 5' to the deletion, showing that, in vivo, the 5' side is strongly favored by recombination. Further results suggest that exchanges occurring from 5' to 3' relative to the donor strand are more efficient than in the opposite direction, thus accounting for the 5' preference.


Vinorelbine Plus Gemcitabine or Cisplatin as First-line Treatment of HER2-negative Advanced Breast Cancer.
To evaluate the efficacy and toxicity of vinorelbine and gemcitabine (NG) versus vinorelbine and cisplatin (NP) in anthracycline- and taxane-pretreated patients with HER2-negative advanced breast cancer. Patients were randomly assigned on a 1:1 schedule to receive no more than six cycles of NG or NP. Dosing for the NG group was 25 mg/m2 vinorelbine and 1,000 mg/m2 gemcitabine, given on days 1 and 8 every 3 weeks; for the NP group,25 mg/m2 vinorelbine was given on days 1 and 8, and 75 mg/m2 cisplatin was given on day 1 every 3 weeks. The primary endpoint was disease control rate (DCR). The secondary endpoints were the progression-free survival (PFS), overall survival (OS) and safety. The full analysis set comprised of 37 patients receiving NG and 37 receiving NP. The DCR was 70.3% with NG and 64.9% with NP (p=0.619). Median PFS were 7 months (95% CI=5.88-8.12) and 6 months (95% CI=5.29-6.71) respectively in NG and NP group [hazard ratio (HR)=1.696; 95% confidence interval (CI)=0.73-3.93; p=0.217)]. Corresponding median OS was 18 (95% CI=10.35-13.65) months and 12 (95% CI=15.83-20.17) months (HR=1.219; 95% CI=0.67-2.23; p=0.521). For adverse events, neutropenia and nausea/vomiting were milder in the NG group than in the NP group (all p<0.05). Although no significant differences were observed in terms of DCR, PFS and OS, with milder toxicity, NG appeared to be a more valuable first-line treatment regimen than NP in anthracycline- and taxane-pretreated patients with HER2-negative advanced breast cancer.


Simplification and its consequences in biological modelling: conclusions from a study of calcium oscillations in hepatocytes.
Systems Biology requires that biological modelling is scaled up from small components to system level. This can produce exceedingly complex models, which obscure understanding rather than facilitate it. The successful use of highly simplified models would resolve many of the current problems faced in Systems Biology. This paper questions whether the conclusions of simple mathematical models of biological systems are trustworthy. The simplification of a specific model of calcium oscillations in hepatocytes is examined in detail, and the conclusions drawn from this scrutiny generalized. We formalize our choice of simplification approach through the use of functional 'building blocks'. A collection of models is constructed, each a progressively more simplified version of a well-understood model. The limiting model is a piecewise linear model that can be solved analytically. We find that, as expected, in many cases the simpler models produce incorrect results. However, when we make a sensitivity analysis, examining which aspects of the behaviour of the system are controlled by which parameters, the conclusions of the simple model often agree with those of the richer model. The hypothesis that the simplified model retains no information about the real sensitivities of the unsimplified model can be very strongly ruled out by treating the simplification process as a pseudo-random perturbation on the true sensitivity data. We conclude that sensitivity analysis is, therefore, of great importance to the analysis of simple mathematical models in biology. Our comparisons reveal which results of the sensitivity analysis regarding calcium oscillations in hepatocytes are robust to the simplifications necessarily involved in mathematical modelling. For example, we find that if a treatment is observed to strongly decrease the period of the oscillations while increasing the proportion of the cycle during which cellular calcium concentrations are rising, without affecting the inter-spike or maximum calcium concentrations, then it is likely that the treatment is acting on the plasma membrane calcium pump.


Ranking docking poses by graph matching of protein-ligand interactions: lessons learned from the D3R Grand Challenge 2.
A novel docking challenge has been set by the Drug Design Data Resource (D3R) in order to predict the pose and affinity ranking of a set of Farnesoid X receptor (FXR) agonists, prior to the public release of their bound X-ray structures and potencies. In a first phase, 36 agonists were docked to 26 Protein Data Bank (PDB) structures of the FXR receptor, and next rescored using the in-house developed GRIM method. GRIM aligns protein-ligand interaction patterns of docked poses to those of available PDB templates for the target protein, and rescore poses by a graph matching method. In agreement with results obtained during the previous 2015 docking challenge, we clearly show that GRIM rescoring improves the overall quality of top-ranked poses by prioritizing interaction patterns already visited in the PDB. Importantly, this challenge enables us to refine the applicability domain of the method by better defining the conditions of its success. We notably show that rescoring apolar ligands in hydrophobic pockets leads to frequent GRIM failures. In the second phase, 102 FXR agonists were ranked by decreasing affinity according to the Gibbs free energy of the corresponding GRIM-selected poses, computed by the HYDE scoring function. Interestingly, this fast and simple rescoring scheme provided the third most accurate ranking method among 57 contributions. Although the obtained ranking is still unsuitable for hit to lead optimization, the GRIM-HYDE scoring scheme is accurate and fast enough to post-process virtual screening data.


Everything Is Everywhere: Physiological Responses of the Mediterranean Sea and Eastern Pacific Ocean Epiphyte Cobetia Sp. to Varying Nutrient Concentration.
Bacteria are essential in the maintenance and sustainment of marine environments (e.g., benthic systems), playing a key role in marine food webs and nutrient cycling. These microorganisms can live associated as epiphytic or endophytic populations with superior organisms with valuable ecological functions, e.g., seagrasses. Here, we isolated, identified, sequenced, and exposed two strains of the same species (i.e., identified as Cobetia sp.) from two different marine environments to different nutrient regimes using batch cultures: (1) Cobetia sp. UIB 001 from the endemic Mediterranean seagrass Posidonia oceanica and (2) Cobetia sp. 4B UA from the endemic Humboldt Current System (HCS) seagrass Heterozostera chilensis. From our physiological studies, both strains behaved as bacteria capable to cope with different nutrient and pH regimes, i.e., N, P, and Fe combined with different pH levels, both in long-term (12 days (d)) and short-term studies (4 d/96 h (h)). We showed that the isolated strains were sensitive to the N source (inorganic and organic) at low and high concentrations and low pH levels. Low availability of phosphorus (P) and Fe had a negative independent effect on growth, especially in the long-term studies. The strain UIB 001 showed a better adaptation to low nutrient concentrations, being a potential N2-fixer, reaching higher growth rates (μ) than the HCS strain. P-acquisition mechanisms were deeply investigated at the enzymatic (i.e., alkaline phosphatase activity, APA) and structural level (e.g., alkaline phosphatase D, PhoD). Finally, these results were complemented with the study of biochemical markers, i.e., reactive oxygen species (ROS). In short, we present how ecological niches (i.e., MS and HCS) might determine, select, and modify the genomic and phenotypic features of the same bacterial species (i.e., Cobetia spp.) found in different marine environments, pointing to a direct correlation between adaptability and oligotrophy of seawater.


End-of-life and palliative care of patients on maintenance hemodialysis treatment: a focus group study.
Despite complex illness trajectories and a high symptom burden, palliative care has been sub-optimal for patients with end-stage kidney disease and hemodialysis treatment who have a high rate of hospitalization and intensive care towards end of life. There is a growing awareness that further development of palliative care is required to meet the needs of these patients and their family members. In this process, it is important to explore healthcare professionals' views on provision of care. The aim of this study was therefore to describe nurses' and physicians' perspectives on end-of-life and palliative care of patients treated with maintenance hemodialysis. Four focus group interviews were conducted with renal nurses (17) and physicians (5) in Sweden. Qualitative content analysis was used to analyze data. Participants were committed to giving the best possible care to their patients, but there were challenges and barriers to providing quality palliative care in nephrology settings. Professionals described palliative care as end-of-life care associated with hemodialysis withdrawal or palliative dialysis, but also identified care needs and possibilities that are in line with an earlier integrated palliative approach. This was perceived as complex from an organizational point of view. Participants identified challenges related to coordination of care and different perspectives on care responsibilities that impacted symptom management and patients' quality of life. Communication issues relating to the provision of palliative care were revealed where the hemodialysis setting was regarded as an impediment, and personal and professional experiences, beliefs and knowledge were considered of major importance. Nurses and physicians identified a need for the improvement of both late and earlier palliative care approaches. The results highlighted a requirement for and possibilities of training, counselling and support of health care professionals in the dialysis context. Further, multi-professional palliative care collaborations should be developed to improve the coordination and organization of end-of-life and palliative care of patients and their family members. A climate allowing conversations about advance care planning throughout the illness trajectory may facilitate the gradual integration of palliative care alongside life-prolonging treatment for improved support of patients and families.


Experimental study of plasmid TGF-beta 1 DNA gene transfer with lipofectamine into rabbit corneal epithelial cells in vitro.
To investigate whether the TGF-beta 1 plasmid DNA carried by lipofectamine could be introduced into cultured rabbit corneal epithelial cells, specific expression of the plasmid pMAM TGF-beta 1 in the cultured corneal epithelial cells was studied. Two days after 12 h of transfection of pMAMT-GF-beta 1 mediated by lipofectamine into the cultured corneal epithelial cells, the TGF-beta 1 protein expression specific for pMAMTGF-beta 1 in the cells was detected by means of immunohistochemical staining and the positive rate was 23.37%. The results suggested that foreign plasmid DNA could be effectively delivered into cultured rabbit corneal epithelial cells by means of lipofectamine, and this will provide a promising method of studying TGF-beta 1 on the mechanism of physiology and pathology concerned with corneal epithelial cells.


SolRgene: an online database to explore disease resistance genes in tuber-bearing Solanum species.
The cultivated potato (Solanum tuberosum L.) is an important food crop, but highly susceptible to many pathogens. The major threat to potato production is the Irish famine pathogen Phytophthora infestans, which causes the devastating late blight disease. Potato breeding makes use of germplasm from wild relatives (wild germplasm) to introduce resistances into cultivated potato. The Solanum section Petota comprises tuber-bearing species that are potential donors of new disease resistance genes. The aim of this study was to explore Solanum section Petota for resistance genes and generate a widely accessible resource that is useful for studying and implementing disease resistance in potato. The SolRgene database contains data on resistance to P. infestans and presence of R genes and R gene homologues in Solanum section Petota. We have explored Solanum section Petota for resistance to late blight in high throughput disease tests under various laboratory conditions and in field trials. From resistant wild germplasm, segregating populations were generated and assessed for the presence of resistance genes. All these data have been entered into the SolRgene database. To facilitate genetic and resistance gene evolution studies, phylogenetic data of the entire SolRgene collection are included, as well as a tool for generating phylogenetic trees of selected groups of germplasm. Data from resistance gene allele-mining studies are incorporated, which enables detection of R gene homologs in related germplasm. Using these resources, various resistance genes have been detected and some of these have been cloned, whereas others are in the cloning pipeline. All this information is stored in the online SolRgene database, which allows users to query resistance data, sequences, passport data of the accessions, and phylogenic classifications. Solanum section Petota forms the basis of the SolRgene database, which contains a collection of resistance data of an unprecedented size and precision. Complemented with R gene sequence data and phylogenetic tools, SolRgene can be considered the primary resource for information on R genes from potato and wild tuber-bearing relatives.


[The in vitro effect of ascorbic acid on sodium nitrite intoxication].
The in vitro effect of ascorbic acid on the content of sodium nitrite in relation to pH and time of incubation and also effect of sodium nitrite on the total content of vitamin C as a sum of both reduced and oxidized forms was demonstrated. Ascorbic acid in respective quantities of 60 mg, 120 mg, 180 mg and sodium nitrite in quantity of 12 mg were added to the in vitro simulated stomach content at pH 1.5, 2.5, 3.5. The control samples were tested without addition of ascorbic acid. The Griess method for determination of nitrite and the Tillmans method with Pijanowski's modification were used for the determination of vitamin C. It was found that the choice of lower pH significantly decreased the level of nitrite in the control samples produced the disappearance of nitrite increased depending on the quantity of ascorbic acid added to the sample and on pH. The total content of vitamin C did not change after adding of sodium nitrite to the samples. The performed studies suggested a positive effect of ascorbic acid decreasing of the nitrite level.


Time-resolved dynamic computational modeling of human EEG recordings reveals gradients of generative mechanisms for the MMN response.
Despite attempts to unify the different theoretical accounts of the mismatch negativity (MMN), there is still an ongoing debate on the neurophysiological mechanisms underlying this complex brain response. On one hand, neuronal adaptation to recurrent stimuli is able to explain many of the observed properties of the MMN, such as its sensitivity to controlled experimental parameters. On the other hand, several modeling studies reported evidence in favor of Bayesian learning models for explaining the trial-to-trial dynamics of the human MMN. However, direct comparisons of these two main hypotheses are scarce, and previous modeling studies suffered from methodological limitations. Based on reports indicating spatial and temporal dissociation of physiological mechanisms within the timecourse of mismatch responses in animals, we hypothesized that different computational models would best fit different temporal phases of the human MMN. Using electroencephalographic data from two independent studies of a simple auditory oddball task (n = 82), we compared adaptation and Bayesian learning models' ability to explain the sequential dynamics of auditory deviance detection in a time-resolved fashion. We first ran simulations to evaluate the capacity of our design to dissociate the tested models and found that they were sufficiently distinguishable above a certain level of signal-to-noise ratio (SNR). In subjects with a sufficient SNR, our time-resolved approach revealed a temporal dissociation between the two model families, with high evidence for adaptation during the early MMN window (from 90 to 150-190 ms post-stimulus depending on the dataset) and for Bayesian learning later in time (170-180 ms or 200-220ms). In addition, Bayesian model averaging of fixed-parameter models within the adaptation family revealed a gradient of adaptation rates, resembling the anatomical gradient in the auditory cortical hierarchy reported in animal studies.


Computer analysis of orbital fat and muscle volumes in Graves ophthalmopathy.
A technique using special computer programs and data from high-resolution computed tomographic scans has been developed that provides accurate and reproducible volume measurements for muscle and fat in the orbit. Normal values for retrobulbar fat and muscle were established in 19 adults. Nineteen patients with Graves exophthalmopathy were then studied, and a spectrum of change was found that included varying degrees of increased muscle volume or increased volume of both muscle and fat. Further categorization of these changes related to clinical characteristics should lead to a better understanding of the mechanism for ophthalmopathy in Graves disease.


An update on the Society for Immunotherapy of Cancer consensus statement on tumor immunotherapy for the treatment of cutaneous melanoma: version 2.0.
Cancer immunotherapy has been firmly established as a standard of care for patients with advanced and metastatic melanoma. Therapeutic outcomes in clinical trials have resulted in the approval of 11 new drugs and/or combination regimens for patients with melanoma. However, prospective data to support evidence-based clinical decisions with respect to the optimal schedule and sequencing of immunotherapy and targeted agents, how best to manage emerging toxicities and when to stop treatment are not yet available. To address this knowledge gap, the Society for Immunotherapy of Cancer (SITC) Melanoma Task Force developed a process for consensus recommendations for physicians treating patients with melanoma integrating evidence-based data, where available, with best expert consensus opinion. The initial consensus statement was published in 2013, and version 2.0 of this report is an update based on a recent meeting of the Task Force and extensive subsequent discussions on new agents, contemporary peer-reviewed literature and emerging clinical data. The Academy of Medicine (formerly Institute of Medicine) clinical practice guidelines were used as a basis for consensus development with an updated literature search for important studies published between 1992 and 2017 and supplemented, as appropriate, by recommendations from Task Force participants. The Task Force considered patients with stage II-IV melanoma and here provide consensus recommendations for how they would incorporate the many immunotherapy options into clinical pathways for patients with cutaneous melanoma. These clinical guidleines provide physicians and healthcare providers with consensus recommendations for managing melanoma patients electing treatment with tumor immunotherapy.


Cefamandole treatment of pulmonary infection caused by gram-negative rods.
The increasing incidence of pneumonia caused by H. influenza and the problem of beta lactamase production (18% of strains in recent reports) are important considerations in the therapy of pneumonia. An antibiotic that is effective for these strains and other common respiratory pathogens will be useful for the therapy of pneumonia. Cefamandole nafate is a new cephalosporin antibiotic with an antimicrobial spectrum similar to cephalothin with increased activity against Escherichia coli, Proteus spp., Enterobacter spp., and Haemophilus influenzae. Seventeen patients with pneumonia presumed to be due to susceptible gram-negative organisms isolated from transtracheal aspirate or sputum were treated with 6 to 8 g/day of parenteral cefamandole nafate. Organisms isolated were Haemophilus influenzae in 6, E. coli in 3, Proteus mirabilis in 2, Klebsiella pneumoniae in 1, Serratia marcescens in 1 and mixed gram-negative rods in 4. The Serratia were resistant (MIC greater than 100 microgram/ml and 50 microgram/ml): other MIC's ranged from 0.2 to 6.2 microgram/ml; median 1.6 microgram/ml. Satisfactory clinical response (improvement in pulmonary function; resolution of infiltrate; decrease in temperature, sputum production and white count) was noted in 13 of 17 patients. Two patients died from their underlying disease. Adverse clinical reactions questionably related to cefamandole included SGOT rises in 3 and rash in one. Serum antibiotic levels were 22.0 to 88.0 microgram/ml (peak) and 1.1 to 12.5 microgram/ml (trough). Sputum levels were 0.27 to 2.5 microgram/ml. Cefamandole appears to be an effective antibiotic for treatment of gram-negative pneumonia caused by susceptible organisms.


Long-term effects of a tube extension on bronchodilator treatment with pressurized aerosol.
The long-term effects of a tube extension to the terbutaline sulphate aerosol were studied in 23 patients. The trial was of open, randomised, cross-over design, and compared the ordinary actuator with the tube extension over two 3-week periods. Lung function before and after bronchodilator was measured at the start of the trial, at the treatment cross-over and at the end of the trial. The mean baseline FEV1 value obtained following 3 weeks of treatment with the tube extension was significantly greater (P less than 0.05) than that following treatment with the ordinary actuator. Treatment with the latter, however, produced significantly greater FEV1 values than at the start of the trial (P less than 0.05). At home, the patients recorded symptoms daily, as well as PEF each morning and evening. With the tube extension, the increase in PEF was significantly larger (P less than 0.05) than with the ordinary actuator. There were no differences with regard to symptom score.


Methotrexate in the treatment of ankylosing spondylitis.
The authors carried out an open prospective study analyzing methotrexate (MTX) efficacy and toxicity in 34 patients with ankylosing spondylitis (AS) for a period of one year. All the patients presented with active axial disease, characterized by inflammatory spinal pain, prolonged morning stiffness, erythrocyte sedimentation rate (ESR) > or = 25 mm, and failure on treatment with non-steroidal anti-inflammatory drugs for a period of more than two years. MTX was taken at a single weekly intramuscular dose of 12.5 mg. Thirty-one patients (91%) concluded treatment. Eighteen patients (53%) were considered responders to MTX; most of them presented peripheral arthritis. Despite clinical improvement, axial measures were unaltered at the end of the study. The mean value of ESR decreased significantly at the end of the treatment (p=0.0001), predominantly in the responders group. Side effects were observed in 23 patients (68%) and included dyspeptic syndrome, transient elevation of liver enzymes, and dizziness. The results of this one year open study suggest that MTX can be an efficient drug in the treatment of AS.


β-PIX controls intracellular viscoelasticity to regulate lung cancer cell migration.
Cancer metastasis occurs via a progress involving abnormal cell migration. Cell migration, a dynamic physical process, is controlled by the cytoskeletal system, which includes the dynamics of actin organization and cellular adhesive organelles, focal adhesions (FAs). However, it is not known whether the organization of actin cytoskeletal system has a regulatory role in the physiologically relevant aspects of cancer metastasis. In the present studies, it was found that lung adenocarcinoma cells isolated from the secondary lung cancer of the lymph nodes, H1299 cells, show specific dynamics in terms of the actin cytoskeleton and FAs. This results in a higher level of mobility and this is regulated by an immature FA component, β-PIX (PAK-interacting exchange factor-β). In H1299 cells, β-PIX's activity was found not to be down-regulated by sequestration onto stress fibres, as the cells did not bundle actin filaments into stress fibres. Thus, β-PIX mainly remained localized at FAs, which allowed maturation of nascent adhesions into focal complexes; this resulted in actin polymerization, increased actin network integrity, changes in the intracellular microrheology at the peripheral of the cell, and cell polarity, which in turn regulated cell migration. Perturbation of β-PIX caused an inhibition of cell migration, including migration velocity, accumulated distance and directional persistence. Our results demonstrate the importance of β-PIX to the regulation of high mobility of lung adenocarcinoma cell line H1299 and that this occurs via regulation of FA dynamics, changes in actin cytoskeleton organization and cell polarity.


[Semax and some glyproline peptides accelerate the healing of acetic ulcers in rats].
Previously we showed that peptides PGP, PG, GP and semax have protective anti-ulcer properties using different models of ulcerogenesis. Semax (MEHFPGP) is a nootropic analogue of adrenocorticotropin 4-7 stabilized by PGP in the C-terminal region. In this study we examined the impact of these peptides on the development and healing of acetic ulcers in rats. The histomorphological and dynamical characteristics of acetic ulcers are most similar to human chronic ulcers. All the peptides were shown to accelerate the process of ulcer cicatrisation in a varying degree (GP semax PG PGP). The dynamics of structural changes in the place of ulcer formation investigated with the use of cytohistological control demonstrated that their anti-ulcer effects can be related to their ability to accelerate ulcer clarification from necrotic tissues and to activate the process of cicatrisation and epithelization. PGP and GP were also shown to reduce the inflammation in the ulcer zone on the fifth day after acetic acid application.


Understanding Suicide Attempts Among Gay Men From Their Self-perceived Causes.
Gay men are at higher risk of suicidality. This paper describes the causes of suicide attempts as perceived by the men themselves and analyzes their impact on severity and recidivism. Mental health surveys conducted among gay men in Geneva, Switzerland, from two probability-based time-space samples in 2007 and 2011, were merged to yield a combined sample N = 762. Suicide ideation, plans, and attempts were assessed, and respondents who had ever attempted suicide answered open questions about perceived causes which were coded and categorized for analysis within the framework of cultural epidemiology. In all, 16.7% of the respondents reported a suicide attempt in their lifetime (59.5% of them with multiple attempts). At their latest attempt, over two thirds asserted intent to die, and half required medical assistance. There was a wide variety of perceived causes, with most individuals reporting multiple causes and many of the most common causes cited at both the first and most recent subsequent attempts. Social/inter-personal problems constitute the most prominent category. Problems with love/relationship and accepting one's homosexuality figure consistently among the top three causes. Whereas the former tend to be associated with weaker intent to die, the latter are associated with the strongest intent to die and reported at multiple attempts. Problems with family are among the most common perceived causes at first attempt but not at the most recent subsequent attempt. Nevertheless, they tend to be related to the strongest intent to die and the greatest medical severity of all the perceived causes. Ten percent of men attempting suicide cited depression as a cause. Although it tended to be associated with weaker intent to die, depression was most likely to be reported at multiple attempts. Respondent-driven assessment yielded both common and idiosyncratic causes of suicide and their distinct effects. Some of these perceived causes are not prominent in the current literature, yet they have important implications for understanding risk and preventing suicide among gay men.


Endoscopic Management of Esophageal Perforations: Who, When, and How?
Esophageal perforations can be spontaneous or iatrogenic. Although they are a rare occurrence, they are associated with a significant morbidity and mortality. Traditionally, management of esophageal perforation consisted of surgery. However, endoscopic management is now emerging as the primary treatment modality and is less invasive and morbid than surgery. Endoscopic modalities include through-the-scope clips (TTS), over-the-scope clips (OTSC), placement of covered stents, and suturing. Suturing can be used for primary closure of the perforation as well as anchoring of stents to prevent migration. Smaller defects (<2 cm) can be closed with clips (TTS or OTSC), whereas larger defects require a stent placement or suturing to achieve closure. If the perforation is associated with a mediastinal collection, drainage is mandatory and can be done via CT-guided percutaneous drainage, surgery, or endoscopic vacuum therapy.


Trace Amine-Associated Receptor 1 Regulation of Methamphetamine Intake and Related Traits.
Continued methamphetamine (MA) use is dependent on a positive MA experience and is likely attenuated by sensitivity to the aversive effects of MA. Bidirectional selective breeding of mice for high (MAHDR) or low (MALDR) voluntary consumption of MA demonstrates a genetic influence on MA intake. Quantitative trait locus (QTL) mapping identified a QTL on mouse chromosome 10 that accounts for greater than 50% of the genetically-determined differences in MA intake in the MAHDR and MALDR lines. The trace amine-associated receptor 1 gene (Taar1) is within the confidence interval of the QTL and encodes a receptor (TAAR1) that modulates monoamine neurotransmission and at which MA serves as an agonist. We demonstrate the existence of a non-functional allele of Taar1 in the DBA/2J mouse strain, one of the founder strains of the selected lines, and show that this non-functional allele co-segregates with high MA drinking and with reduced sensitivity to MA-induced conditioned taste aversion (CTA) and hypothermia. The functional Taar1 allele, derived from the other founder strain, C57BL/6J, segregates with low MA drinking and heightened sensitivity to MA-induced CTA and hypothermia. A role for TAAR1 in these phenotypes is corroborated in Taar1 transgenic mice: Taar1 knockout mice consume more MA and exhibit insensitivity to MA-induced CTA and hypothermia, compared with Taar1 wild-type mice. These are the first data to show that voluntary MA consumption is, in part, regulated by TAAR1 function. Behavioral and physiological studies indicate that TAAR1 function increases sensitivity to aversive effects of MA, and may thereby protect against MA use.


Cloning of the mouse fusin gene, homologue to a human HIV-1 co-factor.
Previous studies have demonstrated that mouse cells do not become infected with HIV-1 despite transfection with human CD4. Recently, a human protein termed "fusin" with characteristics of a seven-transmembrane-spanning receptor was found to be a co-factor required for the entry and fusion of HIV-1 with human CD4-bearing lymphocytes. Thus, cloning of the murine homologue of the human fusin (also termed CXCR-4) gene could provide an important comparative tool for identification of the structures crucial for fusin function. Using degenerate PCR, the mouse homologue of human fusin was cloned from a peritoneal exudate cell cDNA library. The predicted amino acid sequence is 91% identical to human fusin. Twenty-eight of the 37 amino acid differences between mouse and human fusin are located in the ectodomains, suggesting that the intracytoplasmic components that mediate G protein binding and signaling are highly conserved. Northern blot analysis showed a message of 2.2 kb in thymus, spleen, neutrophils, and primary astrocyte cultures. Lymphoid and monocyte cell lines also expressed message for fusin. The coding regions of most chemokine receptors lack introns. In contrast, cloning of genomic DNA for mouse fusin revealed the presence of a 2.3-Kb intron separating the first seven amino acids from the remaining 352 residues. Therefore, the mouse fusin gene has a unique genomic organization compared with other chemokine receptors.


BCG vaccine powder-laden and dissolvable microneedle arrays for lesion-free vaccination.
Live attenuated Bacille Calmette-Guerin (BCG) bacillus is the only licensed vaccine for tuberculosis prevention worldwide to date. It must be delivered intradermally to be effective, which causes severe skin inflammation and sometimes, permanent scars. To minimize the side effects, we developed a novel microneedle array (MNA) that could deliver live attenuated freeze-dried BCG powder into the epidermis in a painless, lesion-free, and self-applicable fashion. The MNA was fabricated with biocompatible and dissolvable hyaluronic acid with a deep cave formed in the basal portion of each microneedle, into which BCG powder could be packaged directly. Viability of BCG vaccine packaged in the caves and the mechanical strength of the powder-laden MNA did not alter significantly before and after more than two months of storage at room temperature. Following insertion of the MNA into the skin, individual microneedle shafts melted away by interstitial fluid from the epidermis and upper dermis, exposing the powder to epidermal tissues. The powder sucked interstitial fluid, dissolved slowly, and diffused into the epidermis in a day against the interstitial fluid influx. Vaccination with BCG-MNA caused no overt skin irritation, in marked contrast to intradermal vaccination that provoked severe inflammation and bruise. While causing little skin irritation, vaccination efficacy of BCG-MNAs was comparable to that of intradermal immunization whether it was evaluated by humoral or cellular immunity. This powder-laden and dissolvable MNA represents a novel technology to sufficiently deliver live attenuated vaccine powders into the skin.


Kynurenic acid is a potent endogenous aryl hydrocarbon receptor ligand that synergistically induces interleukin-6 in the presence of inflammatory signaling.
Inflammatory signaling plays a key role in tumor progression, and the pleiotropic cytokine interleukin-6 (IL-6) is an important mediator of protumorigenic properties. Activation of the aryl hydrocarbon receptor (AHR) with exogenous ligands coupled with inflammatory signals can lead to synergistic induction of IL6 expression in tumor cells. Whether there are endogenous AHR ligands that can mediate IL6 production remains to be established. The indoleamine-2,3-dioxygenase pathway is a tryptophan oxidation pathway that is involved in controlling immune tolerance, which also aids in tumor escape. We screened the metabolites of this pathway for their ability to activate the AHR; results revealed that kynurenic acid (KA) is an efficient agonist for the human AHR. Structure-activity studies further indicate that the carboxylic acid group is required for significant agonist activity. KA is capable of inducing CYP1A1 messenger RNA levels in HepG2 cells and inducing CYP1A-mediated metabolism in primary human hepatocytes. In a human dioxin response element-driven stable reporter cell line, the EC(25) was observed to be 104nM, while in a mouse stable reporter cell line, the EC(25) was 10muM. AHR ligand competition binding assays revealed that KA is a ligand for the AHR. Treatment of MCF-7 cells with interleukin-1beta and a physiologically relevant concentration of KA (e.g., 100nM) leads to induction of IL6 expression that is largely dependent on AHR expression. Our findings have established that KA is a potent AHR endogenous ligand that can induce IL6 production and xenobiotic metabolism in cells at physiologically relevant concentrations.


Znhit1 Regulates p21Cip1 to Control Mouse Lens Differentiation.
The transparency of the ocular lens is essential for refracting and focusing light onto the retina, and transparency is controlled by many factors and signaling pathways. Here we showed a critical role of chromatin remodeler zinc finger HIT-type containing 1 (Znhit1) in maintaining lens transparency. To explore the roles of Znhit1 in lens development, the cre-loxp system was used to generate lens-specific Znhit1 knockout mice (Znhit1Mlr10-Cre; Znhit1 cKO). Morphological changes in mice lenses were examined using hematoxylin and eosin staining. RNA sequencing (RNA-seq) and assay for transposase accessible chromatin using sequencing (ATAC-seq) were applied to screen transcriptome changes. Immunofluorescence staining were performed to assess proteins distribution and terminal deoxynucleotidyl transferase dUTP nick-end labeling staining were used for determining apoptosis. The mRNAs expression was examined by quantitative RT-PCR and proteins expression by Western blot. Lens-specific conditional knockout mice had a severe cataract, microphthalmia phenotype, and seriously abnormal lens fiber cells differentiation. Deletion of Znhit1 in the lens resulted in decreased cell proliferation and increased cell apoptosis of the lens epithelia. ATAC-seq showed that Znhit1 deficiency increased chromatin accessibility of cyclin-dependent kinase inhibitors, including p57Kip2 and p21Cip1, and upregulated the expression of these genes in mRNA and protein levels. And we also showed that loss of Znhit1 lead to lens fibrosis by upregulating the expression of p21Cip1. Our findings suggested that Znhit1 is required for the survival of lens epithelial cells. The loss of Znhit1 leads to the overexpression of p21Cip1, further resulting in lens fibrosis, and impacted the establishment of lens transparency.


Functional variants of the sphingosine-1-phosphate receptor 1 gene associate with asthma susceptibility.
The genetic mechanisms underlying asthma remain unclear. Increased permeability of the microvasculature is a feature of asthma, and the sphingosine-1-phosphate receptor (S1PR1) is an essential participant regulating lung vascular integrity and responses to lung inflammation. We explored the contribution of polymorphisms in the S1PR1 gene to asthma susceptibility. A combination of gene resequencing for single nucleotide polymorphism (SNP) discovery, case-control association, functional evaluation of associated SNPs, and protein immunochemistry studies was used. Immunohistochemistry studies demonstrated significantly decreased S1PR1 protein expression in pulmonary vessels in lungs of asthmatic patients compared with those of nonasthmatic subjects (P < .05). Direct DNA sequencing of 27 multiethnic samples identified 39 S1PR1 variants (18 novel SNPs). Association studies were performed based on genotyping results from cosmopolitan tagging SNPs in 3 case-control cohorts from Chicago and New York totaling 1,061 subjects (502 cases and 559 control subjects). The promoter SNP rs2038366 (-1557G/T) was found to be associated with asthma (P = .03) in European Americans. In African Americans an association was found for both asthma and severe asthma for intronic SNP rs3753194 (c.-164+170A/G; P = .006 and P = .040, respectively) and for promoter SNP rs59317557 (-532C/G) with severe asthma (P = .028). Consistent with predicted in silico functionality, alleles of the promoter SNPs rs2038366 (-1557G/T) and rs59317557 (-532C/G) influenced the activity of a luciferase S1PR1 reporter vector in transfected endothelial cells exposed to growth factors (epidermal growth factor, platelet-derived growth factor, and vascular endothelial growth factor) known to be increased in asthmatic airways. These data provide strong support for a role for S1PR1 gene variants in asthma susceptibility and severity.


Strategies to Obtain Encapsulation and Controlled Release of Small Hydrophilic Molecules.
The therapeutic effect of small hydrophilic molecules is limited by the rapid clearance from the systemic circulation or a local site of administration. The unsuitable pharmacokinetics and biodistribution can be improved by encapsulating them in drug delivery systems. However, the high-water solubility, very hydrophilic nature, and low molecular weight make it difficult to encapsulate small hydrophilic molecules in many drug delivery systems. In this mini-review, we highlight three strategies to efficiently encapsulate small hydrophilic molecules and achieve controlled release: physical encapsulation in micro/nanocapsules, physical adsorption via electronic interactions, and covalent conjugation. The principles, advantages, and disadvantages of each strategy are discussed. This review paper could be a guide for scientists, engineers, and medical doctors who want to improve the therapeutic efficacy of small hydrophilic drugs.


Dose calculation models for proton treatment planning using a dynamic beam delivery system: an attempt to include density heterogeneity effects in the analytical dose calculation.
The gantry for proton radiotherapy at the Paul Scherrer Institute (PSI) is designed specifically for the spot-scanning technique. Use of this technique to its full potential requires dose calculation algorithms which are capable of precisely simulating each scanned beam individually. Different specialized analytical dose calculations have been developed, which attempt to model the effects of density heterogeneities in the patient's body on the dose. Their accuracy has been evaluated by a comparison with Monte Carlo calculated dose distributions in the case of a simple geometrical density interface parallel to the beam and typical anatomical situations. A specialized ray casting model which takes range dilution effects (broadening of the spectrum of proton ranges) into account has been found to produce results of good accuracy. This algorithm can easily be implemented in the iterative optimization procedure used for the calculation of the optimal contribution of each individual scanned pencil beam. In most cases an elemental pencil beam dose calculation has been found to be most accurate. Due to the long computing time, this model is currently used only after the optimization procedure as an alternative method of calculating the dose.


BMP signalling permits population expansion by preventing premature myogenic differentiation in muscle satellite cells.
Satellite cells are the resident stem cells of adult skeletal muscle, supplying myonuclei for homoeostasis, hypertrophy and repair. In this study, we have examined the role of bone morphogenetic protein (BMP) signalling in regulating satellite cell function. Activated satellite cells expressed BMP receptor type 1A (BMPR-1A/Alk-3) and contained phosphorylated Smad proteins, indicating that BMP signalling is operating during proliferation. Indeed, exogenous BMP4 stimulated satellite cell division and inhibited myogenic differentiation. Conversely, interfering with the interactions between BMPs and their receptors by the addition of either the BMP antagonist Noggin or soluble BMPR-1A fragments, induced precocious differentiation. Similarly, blockade of BMP signalling by siRNA-mediated knockdown of BMPR-1A, disruption of the intracellular pathway by either Smad5 or Smad4 knockdown or inhibition of Smad1/5/8 phosphorylation with Dorsomorphin, also caused premature myogenic differentiation. BMP signalling acted to inhibit the upregulation of genes associated with differentiation, in part, through regulating Id1. As satellite cells differentiated, Noggin levels increased to antagonise BMP signalling, since Noggin knockdown enhanced proliferation and impeded myoblast fusion into large multinucleated myotubes. Finally, interference of normal BMP signalling after muscle damage in vivo perturbed the regenerative process, and resulted in smaller regenerated myofibres. In conclusion, BMP signalling operates during routine satellite cell function to help coordinate the balance between proliferation and differentiation, before Noggin is activated to antagonise BMPs and facilitate terminal differentiation.


Effect of arsenic trioxide (ATO) on human lung carcinoma PG cell line: ATO induced apoptosis of PG cells and decreased expression of Bcl-2, Pgp.
Arsenic trioxide (ATO) has been established to be an effective agent for treating newly diagnosed and relapsed acute promyelocytic leukemia (APL) patients. Laboratory data suggest that ATO induces apoptosis of hematopoietic or several solid tumor cells. However, to date, its effect on lung carcinoma has not been fully explored. In the present study, we investigated the effect of ATO on human lung carcinoma PG cells in vitro. We found ATO significantly inhibited the proliferation of PG cells in a dose- and time-dependent manner. ATO-induced apoptosis of PG cells was confirmed by the observance of typical morphological changes and detected by the analysis of flow cytometry (FCM). ATO significantly inhibited Bcl-2 and Pgp expression of PG cells by SABC immunohistochemistry and FCM analysis. In conclusion, our findings indicated that ATO induced apoptosis of PG cells and down-regulation of Bcl-2 and Pgp expressions, and these data might provide some theoretical basis for its clinical use in treating lung carcinoma.


Peptidoglycan Endopeptidase Spr of Uropathogenic Escherichia coli Contributes to Kidney Infections and Competitive Fitness During Bladder Colonization.
Uropathogenic E scherichia coli (UPEC) is the most common pathogen of urinary tract infections (UTIs). Antibiotic therapy is the conventional measure to manage such infections. However, the rapid emergence of antibiotic resistance has reduced the efficacy of antibiotic treatment. Given that the bacterial factors required for the full virulence of the pathogens are potential therapeutic targets, identifying such factors may facilitate the development of novel therapeutic strategies against UPEC UTIs. The peptidoglycan (PG) endopeptidase Spr (also named MepS) is required for PG biogenesis in E. coli. In the present study, we found that Spr deficiency attenuated the ability of UPEC to infect kidneys and induced a fitness defect during bladder colonization in a mouse model of UTI. Based on the liquid chromatography (LC)/mass spectrometry (MS)/MS analysis of the bacterial envelope, spr deletion changed the levels of some envelope-associated proteins, suggesting that Spr deficiency interfere with the components of the bacterial structure. Among the proteins, FliC was significantly downregulated in the spr mutant, which is resulted in reduced motility. Lack of Spr might hinder the function of the flagellar transcriptional factor FlhDC to decrease FliC expression. The motility downregulation contributed to the reduced fitness in urinary tract colonization. Additionally, spr deletion compromised the ability of UPEC to evade complement-mediated attack and to resist intracellular killing of phagocytes, consequently decreasing UPEC bloodstream survival. Spr deficiency also interfered with the UPEC morphological switch from bacillary to filamentous shapes during UTI. It is known that bacterial filamentation protects UPEC from phagocytosis by phagocytes. In conclusion, Spr deficiency was shown to compromise multiple virulence properties of UPEC, leading to attenuation of the pathogen in urinary tract colonization and bloodstream survival. These findings indicate that Spr is a potential antimicrobial target for further studies attempting to develop novel strategies in managing UPEC UTIs.


Vitamin B and One-Carbon Metabolite Profiles Show Divergent Associations with Cardiometabolic Risk Markers but not Cognitive Function in Older New Zealand Adults: A Secondary Analysis of the REACH Study.
Vitamin B inadequacies and elevated homocysteine status have been associated with impaired cognitive and cardiometabolic health with aging. There is, however, a scarcity of research investigating integrated profiles of one-carbon (1C) metabolites in this context, including metabolites of interconnected folate, methionine, choline oxidation, and transsulfuration pathways. The study aimed to examine associations between vitamins B and 1C metabolites with cardiometabolic health and cognitive function in healthy older adults, including the interactive effects of Apolipoprotein E-ε4 status. Three hundred and thirteen healthy participants (65-74 y, 65% female) were analyzed. Vitamins B were estimated according to dietary intake (4-d food records) and biochemical status (serum folate and vitamin B12). Fasting plasma 1C metabolites were quantified by liquid chromatography with tandem mass spectrometry. Measures of cardiometabolic health included biochemical (lipid panel, blood glucose) and anthropometric markers. Cognitive function was assessed by the Computerized Mental Performance Assessment System (COMPASS) and Montreal Cognitive Assessment (MoCA). Associations were analyzed using multivariate linear (COMPASS, cardiometabolic health) and Poisson (MoCA) regression modeling. Over 90% of participants met dietary recommendations for riboflavin and vitamins B6 and B12, but only 78% of males and 67% of females achieved adequate folate intakes. Higher serum folate and plasma betaine and glycine concentrations were associated with favorable cardiometabolic markers, whereas higher plasma choline and homocysteine concentrations were associated with greater cardiometabolic risk based on body mass index and serum lipids concentration values (P< 0.05). Vitamins B and homocysteine were not associated with cognitive performance in this cohort, though higher glycine concentrations were associated with better global cognitive performance (P = 0.017), episodic memory (P = 0.016), and spatial memory (P = 0.027) scores. Apolipoprotein E-ε4 status did not modify the relationship between vitamins B or 1C metabolites with cognitive function in linear regression analyses. Vitamin B and 1C metabolite profiles showed divergent associations with cardiometabolic risk markers and limited associations with cognitive performance in this cohort of healthy older adults.


Distal and proximal cis-regulatory elements sense X chromosome dosage and developmental state at the Xist locus.
Developmental genes such as Xist, which initiates X chromosome inactivation, are controlled by complex cis-regulatory landscapes, which decode multiple signals to establish specific spatiotemporal expression patterns. Xist integrates information on X chromosome dosage and developmental stage to trigger X inactivation in the epiblast specifically in female embryos. Through a pooled CRISPR screen in differentiating mouse embryonic stem cells, we identify functional enhancer elements of Xist at the onset of random X inactivation. Chromatin profiling reveals that X-dosage controls the promoter-proximal region, while differentiation cues activate several distal enhancers. The strongest distal element lies in an enhancer cluster associated with a previously unannotated Xist-enhancing regulatory transcript, which we named Xert. Developmental cues and X-dosage are thus decoded by distinct regulatory regions, which cooperate to ensure female-specific Xist upregulation at the correct developmental time. With this study, we start to disentangle how multiple, functionally distinct regulatory elements interact to generate complex expression patterns in mammals.


Medical ethics in laboratory medicine: A review, with an oath for pathologists.
The basic tenets of medical ethics are: autonomy of the patient, beneficence, non-maleficence and justice. These are usually interpreted in the light of the practice of clinical medicine but also apply to pathology and laboratory medicine, a field in which there is often no direct patient contact. We wished to evaluate these basic tenets of medical ethics with respect to laboratory medicine and to provide insights into some of the issues that laboratory physicians, in routine practice and in academia, face on a regular basis. This was done by using the published literature related to the topic of medical ethics, with a special focus on laboratory medicine, as well as the authors' interpretations and opinions, based on their experience. We conclude that the idea of autonomy of the patient or research participant is pertinent with respect to specimens, autopsies and in legal issues such as consent for publication in the media and social media. Beneficence is relevant with respect to laboratory values in reports, financial issues and in research and education. The concept of non-maleficence is important from the point of view of doing no harm, communication with patients and colleagues, reducing/containing error and misdiagnosis in medicine, screening for disease and in over diagnosis. Justice is applicable to issues of distribution of resources and manpower, and their equitable usage. Many of the tenets, however, need to be interpreted in the light of local laws and customs which differ across the world. We conclude with an Oath for pathologists and laboratory physicians. Key words: medical ethics, misconduct, autonomy, beneficence, non-maleficence, justice, informed consent, medical research, oaths.


Implementing Innovative Approaches to Healthcare in a Lower-Middle Income Country: Perspectives from Malawi.
Safe, reliable, and effective healthcare systems are essential for all nations to ensure the health and wellbeing of their citizens. However, this is not always achievable with clinical therapies constantly evolving, resulting in a domino effect of structural, policy and training changes. For low- and middle-income countries (LMICs), implementing change is restricted. It is essential that innovative and realistic solutions are developed, so that effective change can be realised in LMICs. In this report of a global health conference held in July 2019, six perspectives are presented which aim to generate long-term positive change in Malaŵi. Perspective 1: Pneumonia - the BIOTOPE study (BIOmarkers TO diagnose PnEumonia) sought to determine the aetiology of pneumonia in children presenting in primary care. It assessed blood-based markers of bacterial infection as part of a rapid diagnostic approach to better utilise existing resources in Malaŵi. Perspective 2: Cardiovascular - the CARDIA project (CARdiac Dysfunction in Africa) was established to assess clinical and biochemical phenotypes of diabetic patients in Malaŵi. Perspective 3: Asthma - an observational study was conducted to implement a health system strengthening initiative for asthma. The use of locally adapted formularies and protocols with ongoing online mentoring through expert partnerships provided an opportunity to sustainably build capacity. Perspective 4: Sustainable Partnerships - establishing the Malaŵi electronic Health (eHealth) Research Centre, an international hub to develop education, research and innovation for long-term collaboration. Perspective 5: Part-Time PhD Studies - undertaking a part-time PhD within a LMIC provides logistic challenges, but also a number of opportunities for observational research. Perspective 6: Medical electives - an undergraduate elective allows real exposure to global health and facilitates life-long collaborations at an early stage in a medical career. Malaŵi is an under-doctored and resource-poor country. North-South partnerships in Malaŵi should be strengthened with particular emphasis on healthcare innovations, such as eHealth, which allow healthcare problems to be highlighted early while preventative measures are still possible.


Controlled trial of intravenous metronidazole as an adjunct to corticosteroids in severe ulcerative colitis.
A prospective double blind controlled trial was undertaken to examine the role of metronidazole as an adjunct to corticosteroids in the management of severe ulcerative colitis. Thirty nine patients with severe ulcerative colitis were randomised on admission to hospital to receive either intravenous metronidazole 500 mg eight hourly (19 patients) or an identical intravenous placebo (20 patients). The two groups were similar with respect to age, sex, and the extent of colitis. In addition all patients received a standard intravenous regimen consisting of methyl prednisolone 16 mg six hourly and parenteral nutrition together with a twice daily hydrocortisone 100 mg enema. Treatment was continued for five days when the patients were formally assessed. Fourteen of 19 patients (74%) receiving metronidazole and 14/20 (70%) receiving placebo were substantially improved, or in remission at the end of five days. Five patients treated with metronidazole and six with placebo had no improvement and all proceeded to urgent colectomy with no operative mortality. There were three late deaths, one in the metronidazole and two in the placebo group. These results do not support the routine use of intravenous metronidazole in the treatment of severe ulcerative colitis.


[Hygienic significance of studying gaseous discharge during production of polyvinylchloride resin items].
Experimental study demonstrated no vinyl chloride (VC) emanation from polyvinyl-chloride (PVC) resins in environment under indoor temperature (20 degrees). VC emanation was detected during resin heating to 50 degrees C and higher, its concentration increasing along with temperature elevation. Total decomposition of resin samples was observed at 140 degrees C. When analyzing VC emanation from stabilizer-containing compositions it was pointed out that VC concentration even at 180 degrees C was significantly lower than in the experiment with PVC resins. VC was not detected in the work zone air during PVC film manufacture where the temperature of mixture heating didn't exceed 140 degrees C. One could greatly reduce and even completely avoid VC emanation into the work zone air by means of heating temperature limitation according to the lowest technological limit and introduction of appropriate quantity of stabilizers into the mixture. The study showed that at the examined production of PVC films, footwear and cable coating VC content was significantly lower than MAC (5 mg/m3). VC wasn't detected in the most of samples presented.


Reproductive Indices and Oxidative Stress Biomarkers of Male Wistar Rats Prenatally Exposed to Cigarette Smoke.
The negative influence of cigarette smoking on developing fetus is well documented but reports of prenatal cigarette smoking on male reproductive hormones are controversial. However, shortened anogenital distance (AGD) has been established to be an indicator of potential male infertility. We therefore investigated the effects of prenatal exposure to passive cigarette smoke on AGD, reproductive hormones and oxidative stress biomarkers of Wistar rats. Female rats were randomly divided into two groups (n=5) and cohabited with male. Group 1 was exposed to smoke from an idling cigarette from day 1 of gestation till parturition, while Group 2 served as control (no-exposure). Morphometric variables of the litters were recorded on postnatal day 1 (PND1) and at 6th week postnatal life. The male offspring were then sacrificed by cervical dislocation. Testosterone, luteinizing hormone (LH) and follicle stimulating hormone (FSH) were analysed using ELISA. Serum levels of Catalase, sodium dismutase (SOD), malondialdehyde (MDA), lipid profile and liver function biomarkers were examined spectrophotometrically. On PND1, crown rump length and total body length of rats prenatally exposed to cigarette smoke were significantly shorter. Significantly shorter AGD and crown rump length were also observed at 6th week. Testosterone, LH and FSH were not significantly affected. Cigarette smoke exposure significantly decreased Catalase and SOD while MDA increased. Liver function biomarkers, HDL and LDL were not affected but serum levels of total cholesterol and triglyceride significantly increased. The observed decline in AGD and precipitation of oxidative stress by intrauterine cigarette smoke exposure may predispose to male infertility at adulthood.


The inversion, part-whole, and composite effects reflect distinct perceptual mechanisms with varied relationships to face recognition.
Face recognition is thought to rely on specific mechanisms underlying a perceptual bias toward processing faces as undecomposable wholes. This face-specific "holistic processing" is commonly quantified using 3 measures: the inversion, part-whole, and composite effects. Consequently, many researchers assume that these 3 effects measure the same cognitive mechanism(s) and these mechanisms contribute to the wide range of individual differences seen in face recognition ability. We test these assumptions in a large sample (N = 282), with individual face recognition abilities measured by the well-validated Cambridge Face Perception Test. Our results provide little support for either assumption. The small to nonexistent correlations among inversion, part-whole, and composite effects (correlations between -.03 and .28) fail to support the first assumption. As for the second assumption, only the inversion effect moderately predicts face recognition (r = .42); face recognition was weakly correlated with the part-whole effect (r = .25) and not correlated with the composite effect (r = .04). We rule out multiple artifactual explanations for our results by using valid tasks that produce standard effects at the group level, demonstrating that our tasks exhibit psychometric properties suitable for individual differences studies, and demonstrating that other predicted correlations (e.g., between face perception measures) are robust. Our results show that inversion, part-whole, and composite effects reflect distinct perceptual mechanisms, and we argue against the use of the single, generic term holistic processing when referring to these effects. Our results also question the contribution of these mechanisms to individual differences in face recognition. (PsycINFO Database Record


Characterization of alphaT3-1 cells stably transfected with luteininzing hormone beta-subunit complementary deoxyribonucleic acid.
Luteinizing hormone (LH) consists of alpha- and beta-subunits, and synthesis and secretion of LH are regulated by gonadotropin-releasing hormone (GnRH). In order to examine the molecular mechanisms by which GnRH regulates LH secretion, we transfected alphaT3-1 cells with rat LHbeta-subunit cDNA under the control of a constitutive promoter and established a stable cell line of LH2 cells which secreted LH in response to GnRH. Pulsatile and continuous GnRH pretreatments increased gene expression of the alpha-subunit and synthesis of LH, and enhanced the LH secretion by brief treatments with GnRH and 56 mM KCl. The LH secretions were partially blocked by elimination of extracellular Ca2+. GnRH-induced LH secretion was completely inhibited by calphostin C (a protein kinase C inhibitor) and 1 microM wortmannin. In contrast to the GnRH induction, high K+-induced LH secretion was inhibited by KN93, a Ca2+/calmodulin-dependent protein kinase II inhibitor, as well as by 1 microM wortmannin. We also confirmed that activation of cAMP-pathway induced LH secretion, but activation of mitogen-activated protein (MAP) kinase pathway was not involved in LH secretion. These results suggest that GnRH directly regulates LH secretion as well as LHbeta-subunit synthesis, and that LH2 cells are a useful model for the study of LH secretion induced by several secretagogues.


Association of Built Environments With Frequency of Going Outdoors Among Older Community-Dwelling Adults in Japan.
Built environmental factors are important for encouraging outdoor activity among community-dwelling older adults. A total of 214 participants aged 59-94 years were recruited from local cities in Japan. A mixed-effects multivariate linear regression model was conducted for the analysis. Parks that are easy to get to and car availability were positively associated with the number of days per week with any outdoor exposure. Further, the ability to conduct activities of daily living had a significant effect on the use of safe parks and walking areas and the number of days per week with outdoor time. The findings suggest that subjectively assessed neighborhood-built environments, such as readily available safe parks, walking areas, and car availability, are important for encouraging outdoor activity among older community-dwelling adults in Japan. For those with a limited ability to conduct daily activities, the safeness of parks and walking areas are most important.


β-glucosidase, driven by porcine transthyretin promoter, specific expression in the liver of transgenic mice.
Under the background of food security, using non-grain feed instead of corn-soybean-based feed is an effective measure to alleviate the food-feed competition. While, non-grain feeds are often rich in fiber, which cannot be digested by non-ruminants. Producing heterologous enzymes in non-ruminants to improve cellulose utilization rate is a new research strategy by transgenic technology. In this study, porcine transthyretin (TTR) promoter, signal peptide-coding sequence (CDS), Saccharomycopsis fibuligera β-glucosidase gene (BGL1)-CDS, 6×His sequences fragments were fused into pGL3-control vector to generate transgenic vector. Then, transgenic mice were generated by pronuclear microinjection of the linearized expression vectors. Transgenic mice and their offspring were examined by PCR-based genotyping and copy number variation. Results showed that BGL1 was successfully integrated into the mouse genome and transmitted stably. Furthermore, reverse transcription-polymerase chain reaction (RT-PCR), Western blotting, and β-glucosidase activity assay demonstrated that BGL1 was specifically expressed in the liver, and β-glucosidase activity significantly increased. In addition, liver weight index, cellular morphology, and collagen fiber content of the liver showed that exogenous gene insertion did not cause any lesions to live. Taken together, our findings suggest that β-glucosidase driven by TTR promoter was specifically expressed in the liver of transgenic mice.


1H MR spectroscopic characteristics of kernicterus: a possible metabolic signature.
MR examination of infants with kernicterus shows abnormal changes in signal intensity in various parts of the brain, including the globus pallidus and subthalamic regions. The purpose of this study was to retrospectively analyze proton ((1)H) MR spectroscopic data to see if the MR spectroscopic profiles of infants with hyperbilirubinemia and symptoms of kernicterus provide new insights into the pathophysiology of bilirubin neurotoxicity. Six patients aged 3 days to 3 weeks, with hyperbilirubinemia and symptoms of kernicterus underwent MR imaging and MR spectroscopy with a single-voxel point-resolved spectroscopic sequence. The voxel of interest was centered in the basal ganglia with CSF excluded. Quantitative analysis was done by using LCModel. Between-subject comparisons were based on metabolite ratios relative to creatine. Ratios of taurine, glutamate and glutamine, and myoinositol relative to creatine were significantly elevated (P < .001), whereas the ratio of choline to creatine was significantly decreased (P < .001) compared with normal values published for this age group. Lactate levels were not significantly elevated. Kernicterus has a characteristic signature that is detectable on (1)H-MR spectroscopy. Our results are generally in agreement with what is known about bilirubin pathology, and the finding of increased glutamate and glutamine and decreased choline ratios indicates a possible link between hepatic encephalopathy and kernicterus. This observation may help in elucidating the pathophysiology of bilirubin neurotoxicity.


Hepatitis E virus ORF2 protein over-expressed by baculovirus in hepatoma cells, efficiently encapsidates and transmits the viral RNA to naïve cells.
A recombinant baculovirus(vBacORF2) that expressed the full-length ORF2 capsid protein of a genotype 1 strain of hepatitis E virus(HEV) was constructed. Transduction of S10-3 human hepatoma cells with this baculovirus led to large amounts of ORF2 protein production in ~50% of the cells as determined by immune fluorescence microscopy. The majority of the ORF2 protein detected by Western blot was 72 kDa, the size expected for the full-length protein. To determine if the exogenously-supplied ORF2 protein could transencapsidate viral genomes, S10-3 cell cultures that had been transfected the previous day with an HEV replicon of genotype 1 that contained the gene for green fluorescent protein(GFP), in place of that for ORF2 protein, were transduced with the vBacORF2 virus. Cell lysates were prepared 5 days later and tested for the ability to deliver the GFP gene to HepG2/C3A cells, another human hepatoma cell line. FACS analysis indicated that lysates from cell cultures receiving only the GFP replicon were incapable of introducing the replicon into the HepG2/C3A cells whereas ~2% of the HepG2/C3A cells that received lysate from cultures that had received both the replicon and the baculovirus produced GFP. Therefore, the baculovirus-expressed ORF2 protein was able to trans-encapsidate the viral replicon and form a particle that could infect naïve HepG2/C3A cells. This ex vivo RNA packaging system should be useful for studying many aspects of HEV molecular biology.


Total knee arthroplasty in limbs affected by poliomyelitis.
Little information is available regarding the results and complications of total knee arthroplasty in limbs affected by poliomyelitis with severe knee degeneration. We performed a retrospective chart and radiograph review of patients with a history of poliomyelitis involving a limb that subsequently underwent primary total knee arthroplasty between 1970 and 2000. Sixteen total knee arthroplasties were performed in limbs affected by poliomyelitis in fifteen patients. Eleven patients were followed for a minimum of two years, one (two knees) died before the minimum two-year follow-up could be completed, and three were followed for less than two years. No patient was lost to follow-up. There were two periprosthetic fractures, one peroneal nerve palsy, one avulsion of the patellar tendon, and four cases of recurrent instability. These complications were related to the poor bone quality, valgus deformity, patella baja, poor musculature, and attenuated soft tissues commonly found in knees affected by poliomyelitis. Knee Society pain and knee scores were improved postoperatively for all nine knees with a two-year follow-up that had had at least antigravity quadriceps strength prior to surgery. However, Knee Society function scores remained at 0 or worsened for six of the eleven knees followed for at least two years, including those with less than antigravity strength, and four of the nine knees with at least antigravity strength. None of the prostheses loosened. Pain and knee scores improved following total knee arthroplasty in patients with a history of poliomyelitis and antigravity quadriceps strength, but there was less pain relief in patients with less than antigravity quadriceps strength. Recurrence of instability and progressive functional deterioration is possible in all knees affected by poliomyelitis that have undergone total knee replacement, but they appear to occur more commonly in more severely affected knees.


Identification of an Identical de Novo SCAMP5 Missense Variant in Four Unrelated Patients With Seizures and Severe Neurodevelopmental Delay.
Objective: To establish and broaden the phenotypic spectrum of secretory carrier membrane protein (SCAMP5) associated with epilepsy and neurodevelopmental delay. Methods: A Chinese patient was identified at the First Hospital of Peking University, and the three unrelated patients were recruited from two different countries (Italy and United States) through GeneMatcher. SCAMP5 pathogenic variants were identified by whole exome sequencing; clinical data of the patients were retrospectively collected and analyzed. Result: The onset age of seizures was ranged from 6 to 15 months. Patients had different types of seizures, including focal seizures, generalized tonic-clonic seizures and tonic seizure. One patient showed typical autism spectrum disorder (ASD) symptoms. Electroencephalogram (EEG) findings presented as focal or multifocal discharges, sometimes spreading to generalization. Brain magnetic resonance imaging (MRI) abnormalities were present in each patient. Severe intellectual disability and language and motor developmental disorders were found in our patients, with all patients having poor language development and were nonverbal at last follow-up. All but one of the patients could walk independently in childhood, but the ability to walk independently in one patient had deteriorated with age. All patients had abnormal neurological exam findings, mostly signs of extrapyramidal system involvement. Dysmorphic features were found in 2/4 patients, mainly in the face and trunk. All four unrelated patients were found to have the same heterozygous pathogenic SCAMP5 de novo variant (p. Gly180Trp). Conclusion: Epilepsy, severe developmental delay, abnormal neurological exam findings, with or without ASD or variably dysmorphic features and were common in patients with SCAMP5 variant. The onset time and type of seizure varied greatly. The EEG and brain MRI findings were not consistent, but diverse and nonspecific. The motor ability of patients with heterozygous SCAMP5 variant might have a regressive course; language development was more severely affected.


[Analysis of the components of floral scent in Glochidion puberum using gas chromatography-mass spectrometry with dynamic headspace adsorption].
The floral scent plays the important key role in maintaining the obligate pollination mutualism between Glochidion plants and Epicephala moths. In the study, the dynamic headspace adsorption technique was employed to collect the floral scent emitted by Glochidion puberum, gas chromatography coupled with mass spectrometry (GC-MS) was used for the detection and identification of volatile chemical components in headspace samples of flowers from G. puberum. The peak area normalization was used to determine the relative contents of each odour component. The results showed that 45 compounds mainly consisting of monoterpenes and sesquiterpenes were isolated from the floral scent produced by G. puberum. Especially, both linalool (38.06%) and β-elemene (23.84%) were considered as the major scent components of G. puberum. It was speculated that linalool and β-elemene may be the two potential compounds attracting female Epicephala moths. The study provided the basic data for further electroantennographic detection and bioassays to identify the compounds having the actual physiological activity to female Epicephala moths.