Association between Regimen Composition and Treatment Response in Patients with Multidrug-Resistant Tuberculosis: A Prospective Cohort Study.
For treating multidrug-resistant tuberculosis (MDR TB), the World Health Organization (WHO) recommends a regimen of at least four second-line drugs that are likely to be effective as well as pyrazinamide. WHO guidelines indicate only marginal benefit for regimens based directly on drug susceptibility testing (DST) results. Recent evidence from isolated cohorts suggests that regimens containing more drugs may be beneficial, and that DST results are predictive of regimen effectiveness. The objective of our study was to gain insight into how regimen design affects treatment response by analyzing the association between time to sputum culture conversion and both the number of potentially effective drugs included in a regimen and the DST results of the drugs in the regimen. We analyzed data from the Preserving Effective Tuberculosis Treatment Study (PETTS), a prospective observational study of 1,659 adults treated for MDR TB during 2005-2010 in nine countries: Estonia, Latvia, Peru, Philippines, Russian Federation, South Africa, South Korea, Thailand, and Taiwan. For all patients, monthly sputum samples were collected, and DST was performed on baseline isolates at the US Centers for Disease Control and Prevention. We included 1,137 patients in our analysis based on their having known baseline DST results for at least fluoroquinolones and second-line injectable drugs, and not having extensively drug-resistant TB. These patients were followed for a median of 20 mo (interquartile range 16-23 mo) after MDR TB treatment initiation. The primary outcome of interest was initial sputum culture conversion. We used Cox proportional hazards regression, stratifying by country to control for setting-associated confounders, and adjusting for the number of drugs to which patients' baseline isolates were resistant, baseline resistance pattern, previous treatment history, sputum smear result, and extent of disease on chest radiograph. In multivariable analysis, receiving an average of at least six potentially effective drugs (defined as drugs without a DST result indicating resistance) per day was associated with a 36% greater likelihood of sputum culture conversion than receiving an average of at least five but fewer than six potentially effective drugs per day (adjusted hazard ratio [aHR] 1.36, 95% CI 1.09-1.69). Inclusion of pyrazinamide (aHR 2.00, 95% CI 1.65-2.41) or more drugs to which baseline DST indicated susceptibility (aHR 1.65, 95% CI 1.48-1.84, per drug) in regimens was associated with greater increases in the likelihood of sputum culture conversion than including more drugs to which baseline DST indicated resistance (aHR 1.33, 95% CI 1.18-1.51, per drug). Including in the regimen more drugs for which DST was not performed was beneficial only if a minimum of three effective drugs was present in the regimen (aHR 1.39, 95% CI 1.09-1.76, per drug when three effective drugs present in regimen). The main limitation of this analysis is that it is based on observational data, not a randomized trial, and drug regimens varied across sites. However, PETTS was a uniquely large and rigorous observational study in terms of both the number of patients enrolled and the standardization of laboratory testing. Other limitations include the assumption of equivalent efficacy across drugs in a category, incomplete data on adherence, and the fact that the analysis considers only initial sputum culture conversion, not reversion or long-term relapse. MDR TB regimens including more potentially effective drugs than the minimum of five currently recommended by WHO may encourage improved response to treatment in patients with MDR TB. Rapid access to high-quality DST results could facilitate the design of more effective individualized regimens. Randomized controlled trials are necessary to confirm whether individualized regimens with more than five drugs can indeed achieve better cure rates than current recommended regimens.


Tricuspid regurgitation: when is it time for surgery?
Introduction: The interest in tricuspid regurgitation (TR) surgical treatment has grown in the last years, mostly motivated by the trend for early intervention and the development of minimally invasive surgical techniques. Despite this, there is still a lack of evidence regarding when to intervene in functional tricuspid regurgitation outside the context of left-sided valve surgery and when is the best time to approach primary tricuspid regurgitation. Areas covered: Herein, we present the state-of-the-art in tricuspid regurgitation surgical intervention, covering indications, optimum time, surgical options, and outcomes, as well as the role of transcatheter tricuspid valve intervention in the current era. Expert opinion: Current evidence support that the indications and timing of tricuspid valve invasive management should be centered on a range of factors, including patients' characteristics, disease stage, and anatomical considerations. Early intervention, before severe right ventricle dysfunction and pulmonary hypertension, seems to results in better postoperative outcomes. Transcatheter techniques are arising as potential alternatives for inoperable and high-risk patients.


[Treatment of COVID-19 guided by holistic view of traditional Chinese medicine--therapy aimed at both viral and host].
The global outbreak of coronavirus disease 2019(COVID-19) has further spread, and there is an increasing number of confirmed cases in many countries. On February 28, 2020 of Geneva time, the World Health Organization has raised global risk level to the very high level in view of outbreak of COVID-19. Since some patients' condition appeared to deteriorate rapidly after infection of this 2019 novel coronavirus(2019-nCoV), a variety of treatments should be considered. Holistic view and syndrome differentiation are the two characteristics of traditional Chinese medicine(TCM). Therefore, under the guidance of the holistic view, syndrome diffe-rentiation of TCM has achieved good effects in the treatment of COVID-19. This treatment mainly aimed at eliminating pathogens and strengthening overall health, regulating the balance of body and coordinating various of functions of Zangfu organs. In addition, modern medical proposes host-directed therapy(HDT), a strategy aims to interfere with host cell mechanism, enhance immune responses, and reduce exacerbated inflammation. To some extent, the combined application of HDT and antiviral therapy is highly consistent with the therapeutic concept of the holistic view of TCM. Therefore, under the guidance of the holistic view, syndrome differentiation of TCM uses treatments, such as clearing heat, detoxification, relieving asthma, clearing damp and phlegm, together with Lianhua Qingwen Capsules, Maxing Shigan Decoction, and Haoqin Qingdan Decoction under the guidance of these therapeutic methods. These therapeutic methods and prescriptions intervened with both virus and host at the same time in the treatment of COVID-19, which has important implications for the effective clinical treatment of COVID-19.


Lacunar stroke.
Lacunar infarcts or small subcortical infarcts result from occlusion of a single penetrating artery and account for one quarter of cerebral infarctions. Patients with a lacunar infarct usually present with a classical lacunar syndrome (pure motor hemiparesis, pure sensory syndrome, sensorimotor stroke, ataxic hemiparesis or dysarthria-clumsy hand) and, less frequently, an atypical lacunar syndrome. Hypertension and diabetes mellitus are major risk factors for lacunar stroke. Lacunar infarcts show a paradoxical clinical course with a favorable prognosis in the short term, characterized by a low early mortality and reduced functional disability on hospital discharge, but with an increased risk of death, stroke recurrence and dementia in the mid- and long term. Asymptomatic progression of small-vessel disease is a typical feature of the lacunar infarcts. For this reason, lacunar infarction should be regarded as a potentially severe condition rather than a relatively benign disorder and, therefore, lacunar stroke patients require adequate and rigorous management and follow-up. Antiplatelet drugs, careful blood pressure control, the use of statins and modification of lifestyle risk factors are key elements in secondary prevention after lacunar stroke.


LINC00665 enhances tumorigenicity of endometrial carcinoma by interacting with high mobility group AT-hook 1.
Endometrial carcinoma is a frequently diagnosed cancer among females. LncRNAs are reported to be associated with various cancers. Their biological roles in endometrial carcinoma progression is an emerging scientific area. LINC00665 can exert a significant role in many cancers. However, its potential function in endometrial carcinoma is still poorly known. qRT-PCR was carried out to test expression of LINC00665 and HMGA1. Western blot analysis was carried out to detect protein expression of HMGA1. Cell proliferation was evaluated using Cell Counting Kit-8 (CCK-8) and EdU assay. Flow cytometry assay was used to determine cell apoptosis and cell cycle. Wound healing and transwell invasion assay was carried out to test cell migration and invasion. Immunohistochemical staining and HE staining were conducted to assess Ki-67 and tumor growth respectively. Expression of LINC00665 in clinical endometrial carcinoma tissues and cells was obviously up-regulated. Loss of LINC00665 could repress endometrial carcinoma cell viability, induce cell apoptosis and block cell cycle in G1 phase. KLE and HHUA cell migration and invasion ability were depressed by LINC00665 shRNA. Decrease of LINC00665 suppressed endometrial carcinoma tumorigenicity in vivo. RIP assay proved that LINC00665 directly bound with HMGA1 protein. shRNA of HMGA1 obviously restrained endometrial carcinoma cell growth and cell invasion. LINC00665 might promote endometrial carcinoma progression by positively modulating HMGA1.


Computer-based written emotional disclosure: the effects of advance or real-time guidance and moderation by Big 5 personality traits.
Standard written emotional disclosure (WED) about stress, which is private and unguided, yields small health benefits. The effect of providing individualized guidance to writers may enhance WED, but has not been tested. This trial of computer-based WED compared two novel therapist-guided forms of WED - advance guidance (before sessions) and real-time guidance (during sessions, through instant messaging) - to both standard WED and control writing; it also tested Big 5 personality traits as moderators of guided WED. Young adult participants (n = 163) with unresolved stressful experiences were randomized to conditions, had three, 30-min computer-based writing sessions, and were reassessed six weeks later. Contrary to hypotheses, real-time guidance WED had poorer outcomes than the other conditions on several measures, and advance guidance WED also showed some poorer outcomes. Moderator analyses revealed that participants with low baseline agreeableness, low extraversion, or high conscientiousness had relatively poor responses to guidance. We conclude that providing guidance for WED, especially in real-time, may interfere with emotional processing of unresolved stress, particularly for people whose personalities have poor fit with this interactive form of WED.


Regulations of the European Union for mycotoxins in foods.
Mycotoxins are fungal metabolites that are present in a large part of the world food supply and bear potential threat to food safety. The European Union has been working for several years now on the harmonization of some mycotoxin standards for foods based on toxicological evaluation by its Scientific Committee on Food (SCF). These standards will follow the ALARA (as low as reasonably achievable) principle and will be applied to aflatoxins in the first place followed by ochratoxin A, patulin, and several toxins from Fusarium spp. Detailed regulations (limits, methods of sampling, and analysis) for aflatoxins B, G, and M on the commodities at stake have been worked out. At the moment, a variety of interrelated approaches as to the intake and toxicity of ochratoxin A are being discussed to assess the hazards and to determine the associated risk of this toxin. The other toxins like patulin and the ones derived from Fusarium spp. such as zearalenon and fumonisins will be discussed in the years to come.


Sphingosine 1-phosphate counteracts insulin signaling in pancreatic β-cells via the sphingosine 1-phosphate receptor subtype 2.
Glucolipotoxic stress has been identified as a key player in the progression of pancreatic β-cell dysfunction contributing to insulin resistance and the development of type 2 diabetes mellitus (T2D). It has been suggested that bioactive lipid intermediates, formed under lipotoxic conditions, are involved in these processes. Here, we show that sphingosine 1-phosphate (S1P) levels are not only increased in palmitate-stimulated pancreatic β-cells but also regulate β-cell homeostasis in a divergent manner. Although S1P possesses a prosurvival effect in β-cells, an enhanced level of the sphingolipid antagonizes insulin-mediated cell growth and survival via the sphingosine 1-phosphate receptor subtype 2 (S1P2) followed by an inhibition of Akt-signaling. In an attempt to investigate the role of the S1P/S1P2 axis in vivo, the New Zealand obese (NZO) diabetic mouse model, characterized by β-cell loss under high-fat diet (HFD) conditions, was used. The occurrence of T2D was accompanied by an increase of plasma S1P levels. To examine whether S1P contributes to the morphologic changes of islets via S1P2, the receptor antagonist JTE-013 was administered. Most interestingly, JTE-013 rescued β-cell damage clearly indicating an important role of the S1P2 in β-cell homeostasis. Therefore, the present study provides a new therapeutic strategy to diminish β-cell dysfunction and the development of T2D.


Multifaceted, cross-generational costs of hybridization in sibling Drosophila species.
Maladaptive hybridization, as determined by the pattern and intensity of selection against hybrid individuals, is an important factor contributing to the evolution of prezygotic reproductive isolation. To identify the consequences of hybridization between Drosophila pseudoobscura and D. persimilis, we estimated multiple fitness components for F1 hybrids and backcross progeny and used these to compare the relative fitness of parental species and their hybrids across two generations. We document many sources of intrinsic (developmental) and extrinsic (ecological) selection that dramatically increase the fitness costs of hybridization beyond the well-documented F1 male sterility in this model system. Our results indicate that the cost of hybridization accrues over multiple generations and reinforcement in this system is driven by selection against hybridization above and beyond the cost of hybrid male sterility; we estimate a fitness loss of >95% relative to the parental species across two generations of hybridization. Our findings demonstrate the importance of estimating hybridization costs using multiple fitness measures from multiple generations in an ecologically relevant context; so doing can reveal intense postzygotic selection against hybridization and thus, an enhanced role for reinforcement in the evolution of populations and diversification of species.


Time to maximum effect of lansoprazole on gastric pH in normal male volunteers.
The time to maximum inhibition of gastric acidity resulting from repeated oral dosing with lansoprazole 30 mg daily for 7 days was studied in nine healthy male volunteers. Twenty-four hour intragastric pH monitoring was performed before treatment and on days 1, 3, 5 and 7 of dosing with lansoprazole. Blood samples were taken for the estimation of plasma lansoprazole concentrations. Lansoprazole 30 mg increased mean 24-h intragastric pH to 3.57 on day 1 compared with baseline mean pH of 2.11 (P < 0.05). The mean intragastric pH during the morning period (08.00-13.00 h) was significantly higher on days 3, 5 and 7 than on day 1, but no consistent differences between day 1, 3, 5 and 7 were noted for subsequent periods (13.00-18.00, 18.00-21.00 and 23.00-07.00 h). There were no differences in mean pH between days 3, 5 and 7. Intragastric pH was maintained above pH 3 for 54.7, 60.1, 61.9 and 67.4% of the time on days 1, 3, 5 and 7, respectively. Lansoprazole pharmacokinetic parameters did not change with daily dosing. The area under the lansoprazole plasma concentration-time curve correlated with the intragastric pH (P < 0.005). Lansoprazole 30 mg raised intragastric pH significantly from baseline on day 1 to a maximum effect as early as 6 h after the first dose. The degree and duration of acid suppression confirm the usefulness of lansoprazole for the treatment of acid-related disorders.


Pediatric Renal Rickets at a Tertiary Center.
We report clinical and etiological profile of 19 children (10 males) with renal rickets managed in the years 2021-2022. Median (IQR) age of presentation was 60 (18-96) months. The commonest cause was renal tubular acidosis (n=8). Genetic analysis revealed the diagnosis in 83% subjects (5 out of 6 tested).


Recommendation to develop strategies to increase the number of ST-segment-elevation myocardial infarction patients with timely access to primary percutaneous coronary intervention.
Although evidence suggests that primary percutaneous coronary intervention (PCI) is the preferred reperfusion strategy in the majority of patients with ST-segment-elevation myocardial infarction (STEMI), only a minority of patients with STEMI are treated with primary PCI, and of those, only a minority receive the treatment within the recommended 90 minutes after entry into the medical system. Market research conducted by the American Heart Association revealed that those involved in the care of patients with STEMI recognize the multiple barriers that prevent the prompt delivery of primary PCI and agree that it is necessary to develop systems or centers of care that will allow STEMI patients to benefit from primary PCI. The American Heart Association will convene a group of stakeholders (representing the interests of patients, physicians, emergency medical systems, community hospitals, tertiary hospitals, and payers) and quality-of-care and outcomes experts to identify the gaps between the existing and ideal delivery of care for STEMI patients, as well as the requisite policy implications. Working within a framework of guiding principles, the group will recommend strategies to increase the number of STEMI patients with timely access to primary PCI.


Vertical GaN MOSFET Power Devices.
Gallium nitride (GaN) possesses remarkable characteristics such as a wide bandgap, high critical electric field, robust antiradiation properties, and a high saturation velocity for high-power devices. These attributes position GaN as a pivotal material for the development of power devices. Among the various GaN-based devices, vertical GaN MOSFETs stand out for their numerous advantages over their silicon MOSFET counterparts. These advantages encompass high-power device applications. This review provides a concise overview of their significance and explores their distinctive architectures. Additionally, it delves into the advantages of vertical GaN MOSFETs and highlights their recent advancements. In conclusion, the review addresses methods to enhance the breakdown voltage of vertical GaN devices. This comprehensive perspective underscores the pivotal role of vertical GaN MOSFETs in the realm of power electronics and their continual progress.


Portal Vein Recanalization and Transjugular Intrahepatic Portosystemic Shunt Creation for Chronic Portal Vein Thrombosis: Technical Considerations.
Portal vein thrombosis (PVT) is common in cirrhotic patients and presents a challenge at the time of transplant. Owing to the increased posttransplant morbidity and mortality associated with complete PVT, the presence of PVT is a relative contraindication to liver transplantation at many centers. Our group began performing portal vein (PV) recanalization and transjugular intrahepatic portostystemic shunt placement (PVR-TIPS) several years ago to optimize the transplant candidacy of patients with PVT. The procedure has evolved to include transsplenic access to assist with recanalization, which is now our preferred method due to its technical success without significant added morbidity. Here, we describe in detail our approach to PVR-TIPS with a focus on the transsplenic method. The procedure was attempted in 61 patients and was technically successful in 60 patients (98%). After transitioning to transsplenic access to assist with recanalization, the technical success rate has improved to 100%. The recanalized portal vein and TIPS have maintained patency during follow-up, or to the time of transplant, in 55 patients (92%) with a mean follow-up of 16.7 months. In total, 23 patients (38%) have undergone transplant, all of whom received a physiologic anastomosis (end-to-end anastomosis in 22 of 23 patients, 96%). PVR-TIPS placement should be considered as an option for patients with chronic PVT in need of transplantation. Transsplenic access makes the procedure technically straightforward and should be considered as the primary method for recanalization.


Virus strain influenced the interspecies transmission of influenza D virus between calves and pigs.
Influenza D viruses (IDV) belong to a new genus in the family Orthomyxoviridae. IDV is the aetiologic agent of acute, mild respiratory disease in ungulate species with agricultural importance (cattle, pigs, sheep, goats, camels, etc.). Despite the initial isolate being of porcine origin, serological data suggest cattle to be the primary host of IDV. The study aims were twofold: elucidating species-specific replication kinetics of IDV in bovine and porcine hosts and defining the interspecies potential with two different IDV strains. Three calves and three pigs were intranasally inoculated with the prototypic strain D/swine/Oklahoma/1334/2017 or a genetically distinct cattle isolate, D/bovine/Texas/72/2017. Two days following infection, three naïve pigs and three naïve calves were co-housed with inoculated calves and pigs, respectively. The species of IDV origin had no effect on virus replication kinetics in the upper respiratory tract of inoculated calves and pigs; similar shedding profiles were observed for each species and virus. However, interspecies transmission was found to be associated with virus origin species; D/bovine/Texas/72/2017 and D/swine/Oklahoma/1334/2017 were directly transmitted only to contact calves or pigs, respectively. Even so, transmission efficiency was higher for calves compared to pigs. Together, these data show that cattle and pigs are permissive for IDV replication, but IDV transmission may be species dependent. Host-specific mutations likely influenced transmission efficiencies between agriculturally important mammalian species.


Budd--Chiari syndrome and heparin-induced thrombocytopenia.
We report on a patient suffering from Budd--Chiari disease who developed heparin-induced thrombocytopenia preoperatively. Dorsocranial liver resection and hepatoatrial anastomosis were performed with the extracorporeal circulation and perioperative anticoagulation was achieved with r-hirudin. Surprisingly, thrombus formation was observed in the venous reservoir although ACT was 590 s and aPTT 55 s. An additional bolus of hirudin and rinsing the reservoir allowed unproblematic discontinuation of the cardiopulmonary bypass.


Monocrotophos pesticide decreases the plasma levels of total 3,3',5-triiodo-l-thyronine and alters the expression of genes associated with the thyroidal axis in female goldfish (Carassius auratus).
Our recent study showed that monocrotophos (MCP) pesticide disrupted the hypothalamic-pituitary-thyroid (HPT) axis in male goldfish (Carassius auratus); however, the effects of MCP on the thyroid system in female goldfish are remain unclear. In the present study, plasma thyroid hormone (TH) and thyroid-stimulating hormone (TSH) levels were evaluated in female goldfish exposed to 0.01, 0.10, and 1.00 mg/L of 40% MCP-based pesticide for 21 days in a semi-static exposure system. Expression profiles of HPT axis-responsive genes, including transthyretin (ttr), deiodinases (d1, d2, and d3), tshβ, thyrotropin-releasing hormone (trh), and corticotrophin-releasing hormone (crh), were determined. The results indicated that MCP decreased the plasma levels of total 3,3',5-triiodo-l-thyronine (TT3) and the ratio of TT3 to total 3,3',5,5'-l-thyroxine (TT4), and induced alternative expression of TH-related genes. Exposure to 0.01 and 0.10 mg/L MCP pesticide resulted in the up-regulation of ttr mRNA. The reduction of plasma TT3 levels was partly attributed to an increase in the metabolism of T3 in the liver, as revealed by the highly elevated hepatic d1 and d3 mRNA levels in the MCP treatment groups, and the expression of hepatic d3 showed a negative correlation with the plasma TT3/TT4 levels in females. Moreover, the plasma TSH levels were lower in females exposed to 0.01 and 0.10 mg/L MCP pesticide, whereas the up-regulation of tshβ mRNA levels was compensated by the decreased plasma TT3 levels. These results indicated that MCP had the potential to influence several pathways of HPT axis homeostasis in female goldfish.


Use of proteolytic enzymes as an additional tool for trypanosomatid identification.
The expression of proteolytic activities in the Trypanosomatidae family was explored as a potential marker to discriminate between the morphologically indistinguishable flagellates isolated from insects and plants. We have comparatively analysed the proteolytic profiles of 19 monoxenous trypanosomatids (Herpetomonas anglusteri, H. samuelpessoai, H. mariadeanei, H. roitmani, H. muscarum ingenoplastis, H. muscarum muscarum, H. megaseliae, H. dendoderi, Herpetomoas sp., Crithidia oncopelti, C. deanei, C. acanthocephali, C. harmosa, C. fasciculata, C. guilhermei, C. luciliae, Blastocrithidia culicis, Leptomonas samueli and Lept. seymouri) and 4 heteroxenous flagellates (Phytomonas serpens, P. mcgheei, Trypanosoma cruzi and Leishmania amazonensis) by in situ detection of enzyme activities on sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE ) containing co-polymerized gelatine as substrate, in association with specific proteinase inhibitors. All 23 trypanosomatids expressed at least 1 acidic proteolytic enzyme. In addition, a characteristic and specific pattern of cell-associated metallo and/or cysteine proteinases was observed, except for the similar profiles detected in 2 Herpetomonas (H. anglusteri and H. samuelpessoai) and 3 Crithidia (C. fasciculata, C. guilhermei and C. luciliae) species. However, these flagellates released distinct secretory proteinase profiles into the extracellular medium. These findings strongly suggest that the association of cellular and secretory proteinase pattern could represent a useful marker to help trypanosomatid identification.


Cortical modulation of whole body movements in brain-damaged patients.
The aim of the present study was to assess whether the consistent bias in reproducing distances and lengths on visual tasks that characterizes hemispatial neglect is also present when whole body displacements have to be calculated and reproduced. Two different experiments were proposed to participants with right brain lesions and neglect syndrome (RN+), right (RN-) and left brain lesions (LN-) without neglect and to participants without brain damage (C). In Experiment 1, participants actively reproduced passive linear displacements in the same or in a different direction. This task could be performed using only vestibular and somatosensory input, since no relevant visual input was available. In Experiment 2, relevant visual information had to be integrated with vestibular and somatosensory information in order to make the active reproduction. In Experiment 1, all brain-damaged groups reproduced the horizontal displacements similarly to the control group and without any spatial asymmetry. In Experiment 2, when vestibular, somatosensory and visually remembered information was required to produce an integrated mental representation, RN+ processed contralesional displacements differently from ipsilesional ones. Rectilinear displacements of the whole body in space were not affected by focal left and right brain lesions, suggesting that the computation of nonvisual information can be accomplished by brain structures different from those involved in spatial visual processes. However, when body displacements in space required a mental representation based on visual and nonvisual sensory information, a significant asymmetry appeared only in patients with hemispatial neglect. Some attempts are made to identify the neural substrates involved in this integration.


Adult smokers' receptivity to a television advert for electronic nicotine delivery systems.
The aim of the present work was to examine adult smokers' awareness of and receptivity to an electronic nicotine delivery system (ENDS) television advert, and whether viewing the advert influenced urge to smoke and intention to try ENDS. A television advert for ENDS brand blu eCigs was shown to an online convenience sample of 519 Florida adult smokers. We measured current smokers' awareness of and receptivity to the advert, and whether seeing the advert influenced their thoughts about smoking or quitting, urge to smoke and intention to try ENDS. Results were stratified by prior ENDS use. Approximately 62.3% of current smokers were aware of the advert. Smokers found the advert informative (73.8%), attention grabbing (67.5%) and innovative (64.5%), with prior ENDS users rating the advert more favourably than non-users. Seeing the advert elicited an urge to smoke (mean 42.1, SD=1.9) and thoughts about smoking cigarettes (75.8%) as well as quitting (74.6%). Prior END users were significantly more likely than non-users to report thinking about smoking cigarettes after seeing the advert (P<0.05). Most smokers said ENDS were 'made for people like them' (88.6%) and they would try ENDS in the future (66.0%). Smokers are receptive to ENDS television adverts and report intention to try ENDS after viewing the advert. Future studies should monitor ENDS advertising and examine how exposure to ENDS adverts influences smokers' use of ENDS, dual use with cigarettes and cessation behaviour.


Expression of the RNA component of human telomerase in adult testicular germ cell neoplasia.
During human development, telomerase is repressed in most somatic cells, whereas it is maintained in male germline cells. Reactivation of telomerase has been associated with somatic cancers. To the authors' knowledge, the role of telomerase in germ cell derived malignancies has not previously been evaluated. A radioactive in situ hybridization method was used to study the expression of the RNA component of human telomerase (hTR) in 22 cases of adult testicular germ cell neoplasia encompassing all major histomorphologic types. For precise cell identification, hTR in situ hybridization was combined with immunohistochemistry in select cases. Testicular germ cell tumors showed differential expression of hTR. The highest level of expression was seen in embryonal carcinoma. Seminoma and unclassified intratubular germ cell neoplasia exhibited moderate levels of expression. Yolk sac tumor was characterized by a range of expression, which mirrored its morphologic variation. Immature teratoma recapitulated the down-regulation of telomerase manifested during human embryogenesis. Mature teratoma represented the adult pattern of somatic repression. Notably, choriocarcinoma showed modest expression. The expression of spermatocytic seminoma was intermediate between that of classic seminoma and embryonal carcinoma. No difference in expression was evident between matching intratubular and invasive components. In nonneoplastic testis, hTR expression was down-regulated during spermatogenesis and was absent in spermatozoa. Expression was negligible in rete testis and interstitial Leydig cells, and low in epididymis. Unexpectedly, Sertoli cells, which are testicular accessory somatic cells, displayed the most intense expression observed in this study. In testicular germ cell tumors of young adults (and during spermatogenesis), hTR expression is down-regulated with differentiation, irrespective of the aggressiveness of the tumors. Spermatocytic seminoma, regarded as a low grade malignancy, shows moderately intense expression.


Microbiome-related aspects of locust density-dependent phase transition.
Locust plagues are a notorious, ancient phenomenon. These swarming pests tend to aggregate and perform long migrations, decimating cultivated fields along their path. When population density is low, however, the locusts will express a cryptic, solitary, non-aggregating phenotype that is not considered a pest. Although the transition from the solitary to the gregarious phase has been well studied, associated shifts in the locust's microbiome have yet to be addressed. Here, using 16S rRNA amplicon sequencing, we compared the bacterial composition of solitary desert locusts before and after a phase transition. Our findings revealed that the microbiome is altered during the phase transition, and that a major aspect of this change is the acquisition of Weissella (Firmicutes). Our findings led us to hypothesize that the locust microbiome plays a role in inducing aggregation behaviour, contributing to the formation and maintenance of a swarm. Employing a mathematical model, we demonstrate the potential evolutionary advantage of inducing aggregation under different conditions; specifically, when the aggregation-inducing microbe exhibits a relatively high horizontal transmission rate. This is the first report of a previously unknown and important aspect of locust phase transition, demonstrating that the phase shift includes a shift in the gut and integument bacterial composition.


Intensifying platelet inhibition with tirofiban in poor responders to aspirin, clopidogrel, or both agents undergoing elective coronary intervention: results from the double-blind, prospective, randomized Tailoring Treatment with Tirofiban in Patients Showing Resistance to Aspirin and/or Resistance to Clopidogrel study.
Inhibition of platelet aggregation after aspirin or clopidogrel intake varies greatly among patients, and previous studies have suggested that poor response to oral antiplatelet agents may increase the risk of thrombotic events, especially after coronary angioplasty. Whether this reflects suboptimal platelet inhibition per se, which might benefit from more potent antiplatelet agents such as tirofiban, is unknown. We screened 1277 patients to enroll 93 aspirin, 147 clopidogrel, and 23 dual poor responders, based on a point-of-care assay, who underwent elective coronary angioplasty at 10 European sites for stable or low-risk unstable coronary artery disease. Patients were randomly assigned in a double-blind manner to receive either tirofiban (n=132) or placebo (n=131) on top of standard aspirin and clopidogrel therapy. The primary end point, consisting of troponin I/T elevation at least 3 times the upper limit of normal, was attained in 20.4% (n=27) in the tirofiban group compared with 35.1% (n=46) in the placebo group (relative risk, 0.58; 95% confidence interval, 0.39 to 0.88; P=0.009). The rate of major adverse cardiovascular events within 30 days in the tirofiban group also was reduced (3.8% versus 10.7%; P=0.031). The overall incidence of bleeding was low, likely explained by a substantial use of the transradial approach, and did not differ between the 2 groups. In low-risk patients according to clinical presentation who had poor responsiveness to standard oral platelet inhibitors via a point-of-care assay, intensified platelet inhibition with tirofiban lowers the incidence of myocardial infarction after elective coronary intervention.


Thyroid hormone: half-sites and insights. Molecular Approaches to the Study of Thyroid hormone Action: a Keystone Symposium, Tamarron, CO, USA, March 8-14, 1991.
As is evident from the range of topics addressed in this review, T3 has important and interesting effects on virtually every biological system in higher organisms. Many of these effects are beginning to be understood at the molecular level, while others are not far behind. It is striking that the diversity of biological activities of T3 is reflected in a broad spectrum of molecular mechanisms. Even within the relatively circumscribed paradigm of receptor activation by hormone binding, a surprisingly large number of possible interactions and complications has been discovered. This richness of alternative interactions leads directly to complex and unanticipated regulatory possibilities. A major strength of this meeting was that it encompassed the broad range of T3 effects and thereby was able to provide something new for every attendee. From this perspective, it is easy to predict that study of T3 will continue to provide new and surprising insights into major biological questions for years to come.


Two-peak and three-peak optimal complex networks.
A central issue in complex networks is tolerance of random failures and intentional attacks. Current literature emphasizes the dichotomy between networks with a power-law node connectivity distribution, which are robust to random failures but fragile to targeted attacks, versus networks with an exponentially decaying connectivity distribution, which are less tolerant to failures but more resilient to attacks. We prove analytically that the optimal network configuration under a classic measure of robustness is altogether different from both of the above: in all cases, failure and/or attack, there are no more than three distinct node connectivities in the optimal network.


Rate-distortion modeling for multiscale binary shape coding based on Markov random fields.
The purpose of this paper it to explore the relationship between the rate-distortion characteristics of multiscale binary shape and Markov random field (MRF) parameters. For coding, it is important that the input parameters that will be used to define this relationship be able to distinguish between the same shape at different scales, as well as different shapes at the same scale. We consider an MRF model, referred to as the Chien model, which accounts for high-order spatial interactions among pixels. We propose to use the statistical moments of the Chien model as input to a neural network to accurately predict the rate and distortion of the binary shape when coded at various scales.


An assessment of the efficiency of PrPsc detection in rectal mucosa and third-eyelid biopsies from animals infected with scrapie.
In classical scrapie, detection of PrPsc on lymphoreticular system is used for the in vivo and post mortem diagnosis of the disease. However, the sensitivity of this methodology is not well characterised because the magnitude and duration of lymphoid tissue involvement can vary considerably. The aim of the present study was to evaluate the efficiency of detecting PrPsc in rectal mucosa and third-eyelid biopsies. A total of 474 genetically susceptible sheep and 24 goats from three scrapie infected flocks were included in this study. A sample from rectal mucosa and a sample from third-eyelid lymphoid tissue were collected from each animal. Biopsy samples were fixed in formaldehyde and processed for immunohistochemical examination. Animals with negative biopsy results were studied more closely through a post mortem examination of central nervous and lymphoreticular systems and if there was a positive result, additional biopsy sections were further tested. The sensitivity of rectal mucosa and third-eyelid assays were 36% and 40% respectively on initial examination but increased to 48% and 44% respectively after retesting. The results of this field study show a high percentage of infected animals that do not have detectable levels of PrPsc in the biopsied lymphoid tissue, due mainly to the relatively high number of animals with minimal or no involvement of lymphoid tissue in the pathogenesis of the disease.


Does the use of immunohistochemistry to identify micrometastases provide useful information in the staging of node-negative non-small cell lung carcinomas?
Immunohistochemical studies using epithelial markers have recently been published which identified micrometastases in lymph nodes that had not been found on routine pathological assessment, therefore increasing the accuracy of staging of non-small cell lung cancers. The presence of these micrometastases was associated with reduced survival. We have therefore performed a retrospective immunohistochemical study on all the lymphoid tissue from five lymph node stations (2 hilar, 3 mediastinal) from 49 patients with T1-2, N0 disease. Before immunohistochemistry was undertaken, all slides were reviewed, with the lymph nodes confirmed as negative. In total, 1447 lymph node slices (average 30 per case, 5.9 per lymph node station) were examined, these figures reflecting sectioning of lymph nodes at approximately 3 mm intervals before processing. MNF116, a broad spectrum anti-keratin antibody was then used to look for occult metastases, with adjacent serial sections being examined to ensure that any positively staining cells were detected solely by immunohistochemistry and not through deeper sectioning. In five cases, lymph nodes contained positively staining cells. Two cases proved to be false positives, further immunohistochemistry identifying the cells as benign mesothelial inclusions. In the remaining three cases, positive staining correlated with tumour cells in the adjacent serial sections. Follow-up on 46 of 49 patients revealed recurrence in 27% (actual survival 68%); however all three cases containing tumour cells on immunohistochemistry were free from recurrence. These results suggest that the use of immunohistochemistry adds little useful information above that of thorough routine examination of lymph nodes. They also document that benign mesothelial inclusions within lymph nodes are more frequent than previously reported.


Hyaluronate increases intraocular pressure when used in cataract extraction.
A prospective randomised study of 26 eyes (24 patients) following uncomplicated intracapsular cataract surgery was carried out with three groups in which different agents were used to re-form the anterior chamber: air, seven eyes; sodium hyaluronate, seven eyes; and a combination of hyaluronate plus systemic acetazolamide, 12 eyes. The intraocular pressure (IOP) was measured by Goldmann applanation tonometry 16 hours before surgery and every eight hours after surgery for 72 hours. The IOP doubled in the two hyaluronate-treated groups for the first day following cataract surgery. By contrast, the IOP in the group receiving air initially decreased slightly. Throughout the study the effect of using acetazolamide was never statistically significant. After the first day and a half the differences in IOP among the three treatment groups disappeared, and the pressures returned to their preoperative values.


Signs of Social Class: The Experience of Economic Inequality in Everyday Life.
By some accounts, global economic inequality is at its highest point on record. The pernicious effects of this broad societal trend are striking: Rising inequality is linked to poorer health and well-being across countries, continents, and cultures. The economic and psychological forces that perpetuate inequality continue to be studied, and in this theoretical review, we examine the role of daily experiences of economic inequality-the communication of social class signals between interaction partners-in this process. We theorize that social class signals activate social comparison processes that strengthen group boundaries between the haves and have nots in society. In particular, we argue that class signals are a frequent, rapid, and accurate component of person perception, and we provide new data and analyses demonstrating the accuracy of class signaling in 60-s interactions, Facebook photographs, and isolated recordings of brief speech. We suggest that barriers to the reduction of economic inequality in society arise directly from this class signaling process through the augmentation of class boundaries and the elicitation of beliefs and behaviors that favor the economic status quo.


Impact of dialectical behavior therapy versus community treatment by experts on emotional experience, expression, and acceptance in borderline personality disorder.
Evidence suggests that heightened negative affectivity is a prominent feature of Borderline Personality Disorder (BPD) that often leads to maladaptive behaviors. Nevertheless, there is little research examining treatment effects on the experience and expression of specific negative emotions. Dialectical Behavior Therapy (DBT) is an effective treatment for BPD, hypothesized to reduce negative affectivity (Linehan, 1993a). The present study analyzes secondary data from a randomized controlled trial with the aim to assess the unique effectiveness of DBT when compared to Community Treatment by Experts (CTBE) in changing the experience, expression, and acceptance of negative emotions. Suicidal and/or self-injuring women with BPD (n = 101) were randomly assigned to DBT or CTBE for one year of treatment and one year of follow-up. Several indices of emotional experience and expression were assessed. Results indicate that DBT decreased experiential avoidance and expressed anger significantly more than CTBE. No differences between DBT and CTBE were found in improving guilt, shame, anxiety, or anger suppression, trait, and control. These results suggest that DBT has unique effects on improving the expression of anger and experiential avoidance, whereas changes in the experience of specific negative emotions may be accounted for by general factors associated with expert therapy. Implications of the findings are discussed.


Antagonizing α7 nicotinic receptors with methyllycaconitine (MLA) potentiates receptor activity and memory acquisition.
α7 nicotinic acetylcholine receptors (α7nAChRs) have been targeted to improve cognition in different neurological and psychiatric disorders. Nevertheless, no α7nAChR activating ligand has been clinically approved. Here, we investigated the effects of antagonizing α7nAChRs using the selective antagonist methyllycaconitine (MLA) on receptor activity in vitro and cognitive functioning in vivo. Picomolar concentrations of MLA significantly potentiated receptor responses in electrophysiological experiments mimicking the in vivo situation. Furthermore, microdialysis studies showed that MLA administration substantially increased hippocampal glutamate efflux which is related to memory processes. Accordingly, pre-tetanus administration of low MLA concentrations produced longer lasting potentiation (long-term potentiation, LTP) in studies examining hippocampal plasticity. Moreover, low doses of MLA improved acquisition, but not consolidation memory processes in rats. While the focus to enhance cognition by modulating α7nAChRs lies on agonists and positive modulators, antagonists at low doses should provide a novel approach to improve cognition in neurological and psychiatric disorders.


UK National Audit of chlamydial infection management in sexual health clinics. clinic policies audit.
There was a wide range of activity and chlamydial diagnoses between the 177 clinics that responded. Most (92%) clinics have nucleic acid tests for chlamydial diagnosis. Different practitioners largely share roles in providing advice to patients about partner notification, treatment adherence, safer sex advice and abstinence. Most (97%) clinics have information leaflets about chlamydia, although about 30% of clinics lack leaflets containing information about antibiotics and hormonal contraception. About two-third clinics follow the National Guideline recommended interval for providing a test of cure where this is indicated. Only 18% of clinics routinely ask patients to reattend, with 40% having a policy of no routine follow-up and 62% using telephone or text follow-up. These categories were not mutually exclusive. Most (86%) of the 146 English clinics had a local Chlamydia Screening Programme coordinator for their Primary Care Trust area, although cooperation varies, with cooperation over treatment of 70% and Programme policy of 62%.


Transition Metal Homeostasis in Streptococcus pyogenes and Streptococcus pneumoniae.
Trace metals such as Fe, Mn, Zn and Cu are essential for various biological functions including proper innate immune function. The host immune system has complicated and coordinated mechanisms in place to either starve and/or overload invading pathogens with various metals to combat the infection. Here, we discuss the roles of Fe, Mn and Zn in terms of nutritional immunity, and also the roles of Cu and Zn in metal overload in relation to the physiology and pathogenesis of two human streptococcal species, Streptococcus pneumoniae and Streptococcus pyogenes. S. pneumoniae is a major human pathogen that is carried asymptomatically in the nasopharynx by up to 70% of the population; however, transition to internal sites can cause a range of diseases such as pneumonia, otitis media, meningitis and bacteraemia. S. pyogenes is a human pathogen responsible for diseases ranging from pharyngitis and impetigo, to severe invasive infections. Both species have overlapping capacity with respect to metal acquisition, export and regulation and how metal homeostasis relates to their virulence and ability to invade and survive within the host. It is becoming more apparent that metals have an important role to play in the control of infection, and with further investigations, it could lead to the potential use of metals in novel antimicrobial therapies.


Adolescent Asthma Monitoring: A Preliminary Study of Audio and Spirometry Modalities.
Asthma patients' sleep quality is correlated with how well their asthma symptoms are controlled. In this paper, deep learning techniques are explored to improve forecasting of forced expiratory volume in one second (FEV1) by using audio data from participants and test whether auditory sleep disturbances are correlated with poorer asthma outcomes. These are applied to a representative data set of FEV1 collected from a commercially available sprirometer and audio spectrograms collected overnight using a smartphone. A model for detecting nonverbal vocalizations including coughs, sneezes, sighs, snoring, throat clearing, sniffs, and breathing sounds was trained and used to capture nightly sleep disturbances. Our preliminary analysis found significant improvement in FEV1 forecasting when using overnight nonverbal vocalization detections as an additional feature for regression using XGBoost over using only spirometry data.Clinical relevance- This preliminary study establishes up to 30% improvement of FEV1 forecasting using features generated by deep learning techniques over only spirometry-based features.


The radiomimetic compound streptonigrin induces persistent telomere dysfunction in mammalian cells.
We analyzed the chromosomal aberrations involving telomeres in the progeny of mammalian cells exposed to the radiomimetic compound streptonigrin (SN) in order to determine if this antineoplastic drug induces long-term telomere instability. To this end, rat cells (ADIPO-P2 cell line, derived from adipose cells from Sprague-Dawley rat) were treated with a single concentration of SN (100ng/ml), and chromosomal aberrations were analyzed 18h and 10 and 15 days after treatment by using PNA-FISH with a pan-telomeric probe [Cy3-(CCCTAA)3] to detect (TTAGGG)n repeats. Cytogenetic analysis revealed a higher frequency of telomere dysfunction-related aberrations (additional telomeric FISH signals, extra-chromosomal telomeric FISH signals, and telomere FISH signal loss and duplications) in SN-exposed cultures vs. untreated cultures at every time points analyzed. The yield of SN-induced aberrations remained very similar at 18h, 10 days as well as 15 days after treatment. Thus, our data demonstrate that SN induces persistent telomere dysfunction in mammalian cells. Moreover, we found that the level of telomerase activity in SN-treated cells was significantly lower (up to 77%) than that of untreated control cells at each time points analyzed. This fact suggests that telomerase could be involved in SN-induced telomere dysfunction.


Heterologous expression, purification, crystallization, X-ray analysis and phasing of the acetyl xylan esterase from Bacillus pumilus.
Bacillus pumilus PS213 acetyl xylan esterase (AXE) acts as an accessory enzyme in the plant cell wall hemicellulose biodegradation pathway. It belongs to the carbohydrate esterase family 7 and hydrolyses the ester linkages of the acetyl groups in position 2 and/or 3 of the xylose moieties of the acetylated xylan fragments from hardwood. The enzyme displays activity towards a broad range of acetylated compounds including the antibiotic cephalosporin-C. In this study we report the heterologous expression, purification, physicochemical characterization and crystallization of the recombinant B. pumilus AXE. Remarkable improvement of the crystal quality was achieved by setting up crystallization conditions, at first established using the hanging drop vapor diffusion method, in a micro-batch experiment. Rod-like diffraction quality crystals were obtained using 10% PEG 6000, 0.1 M MES pH 6.0 and a wide range of LiCl concentrations (0.2-1.0 M) as precipitant agent. Two different crystal forms, both belonging to space group P2(1), were characterized, diffracting X-rays to 2.5 and 1.9 angstrom resolution. Successful molecular replacement showed 12 molecules in the asymmetric unit of either crystal forms that are arranged as two doughnut-like hexamers, each one encompassing a local 32 symmetry. A catalytic inactive mutant Ser181Ala of B. pumilus AXE was also engineered, expressed, purified and crystallized for functional and structural studies.


Immunological defects in allergic asthmatic children and working mechanisms of hyposensitization--a review.
Patients with allergic diseases are characterized by the presence of elevated serum IgE and specific IgE antibodies against a variety of environmental allergens, especially house dust, mites and molds in Taiwan. A series of studies on allergic asthmatic children have been conducted to explore the non-immunological and immunological causes for their augmented production of IgE antibody and to explore the working mechanisms of hyposensitization. The results showed that the patients had multiple defects in their defects in their defense mechanisms, including hyper-permeability of mucosae, hyperreactivity of target organs, defective phagocyte functions, and deficient helper and suppressor T-cell functions. Hyposensitization was able to partially correct the immunological aberrations. More over, a newly found lymphokine termed "T-cell growth factor" or "interleukin 2" may be used not only as an indicator for the initiation and termination of hyposensitization, but may also provide a promising tool for the treatment of allergic diseases in the future because of its capability of enhancing and expanding allergen-specific suppressor T cells.


Plasma miRNAs as early biomarkers for detecting hepatocellular carcinoma.
The early detection of hepatocellular carcinoma (HCC) presents a challenge because of the lack of specific biomarkers. Serum/plasma microRNAs (miRNAs) can discriminate HCC patients from controls. We aimed to identify and evaluate HCC-associated plasma miRNAs originating from the liver as early biomarkers for detecting HCC. In this multicenter three-phase study, we first performed screening using both plasma (HCC before and after liver transplantation or liver hepatectomy) and tissue samples (HCC, para-carcinoma and cirrhotic tissues). Then, we evaluated the diagnostic potential of the miRNAs in two case-control studies (training and validation sets). Finally, we used two prospective cohorts to test the potential of the identified miRNAs for the early detection of HCC. During the screening phase, we identified ten miRNAs, eight of which (miR-20a-5p, miR-25-3p, miR-30a-5p, miR-92a-3p, miR-132-3p, miR-185-5p, miR-320a and miR-324-3p) were significantly overexpressed in the HBV-positive HCC patients compared with the HBV-positive cancer-free controls in both the training and validation sets, with a sensitivity of 0.866 and specificity of 0.646. Furthermore, we assessed the potential for early HCC detection of these eight newly identified miRNAs and three previously reported miRNAs (miR-192-5p, miR-21-5p and miR-375) in two prospective cohorts. Our meta-analysis revealed that four miRNAs (miR-20a-5p, miR-320a, miR-324-3p and miR-375) could be used as preclinical biomarkers (pmeta < 0.05) for HCC. The expression profile of the eight-miRNA panel can be used to discriminate HCC patients from cancer-free controls, and the four-miRNA panel (alone or combined with AFP) could be a blood-based early detection biomarker for HCC screening.


PROKR2 missense mutations associated with Kallmann syndrome impair receptor signalling activity.
Kallmann syndrome (KS) combines hypogonadism due to gonadotropin-releasing hormone deficiency, and anosmia or hyposmia, related to defective olfactory bulb morphogenesis. In a large series of KS patients, ten different missense mutations (p.R85C, p.R85H, p.R164Q, p.L173R, p.W178S, p.Q210R, p.R268C, p.P290S, p.M323I, p.V331M) have been identified in the gene encoding the G protein-coupled receptor prokineticin receptor-2 (PROKR2), most often in the heterozygous state. Many of these mutations were, however, also found in clinically unaffected individuals, thus raising the question of their actual implication in the KS phenotype. We reproduced each of the ten mutations in a recombinant murine Prokr2, and tested their effects on the signalling activity in transfected HEK-293 cells, by measuring intracellular calcium release upon ligand-activation of the receptor. We found that all mutated receptors except one (M323I) had decreased signalling activities. These could be explained by different defective mechanisms. Three mutations (L173R, W178S, P290S) impaired cell surface-targeting of the receptor. One mutation (Q210R) abolished ligand-binding. Finally, five mutations (R85C, R85H, R164Q, R268C, V331M) presumably impaired G protein-coupling of the receptor. In addition, when wild-type and mutant receptors were coexpressed in HEK-293 cells, none of the mutant receptors that were retained within the cells did affect cell surface-targeting of the wild-type receptor, and none of the mutant receptors properly addressed at the plasma membrane did affect wild-type receptor signalling activity. This argues against a dominant negative effect of the mutations in vivo.


Radiotherapy and immunotherapy: a synergistic effect in cancer care.
Radiotherapy is an effective treatment modality commonly used in efforts to cure many localised cancers and in the palliation of symptoms in metastatic cancers. Immunotherapy has revolutionised cancer care by increasing the disease control and overall survival of patients in several cancer types; however, the majority of patients do not respond to currently available therapies based on immune checkpoint inhibitors (ICIs). The benefit of those agents is limited to patients who have a pre-existing active immune microenvironment that can be reactivated by ICIs. It is now recognised that radiotherapy does not only directly kill tumour cells but it also changes the tumour microenvironment, enhancing tumour cell recognition by the immune system and, therefore, acting as an in situ vaccine. Radiotherapy increases expression of tumour-associated antigens, causes the release of cytokines, stimulates recruitment of dendritic cells and, most importantly, stimulates the proliferation and priming of cytotoxic CD8+ T cells in the tumour microenvironment. This immunological cascade specifically generates activated T cells able to induce immunogenic cell death directed against cancer cells bearing those antigens. By its ability to overcome some tumour immune escape mechanisms, radiation provides a non-pharmacological and cost-effective approach to potentially improve the systemic response to immune checkpoints inhibitors.


Atypical odontalgia--a diagnostic dilemma.
Atypical odontalgia (AO) is a chronic orofacial pain condition of unclear pathophysiology, often presenting as toothache or pain at an extraction site. Idiopathic, psychogenic, vascular, and neuropathic causes have been proposed. In view of demonstrable somatosensory changes, and responses to management proposed for other forms of neuropathic pain, the best current evidence supports a neuropathic hypothesis. It is proposed that certain individuals with as-yet-undefined genetic vulnerability can develop AO when exposed to certain risk factors, including invasive dental treatment. The diagnosis and treatment of AO can be challenging, but can be aided by a multidisciplinary approach. Two cases of differing complexity are presented in this paper.


The Seasonality of Peripheral Venous Catheter-Related Bloodstream Infections.
Although the seasonality of infectious diseases has been widely reported, the seasonality of peripheral venous catheter-related bloodstream infection (PV-CRBSI) has not been investigated. This study investigated the seasonality of PV-CRBSI and its relationship with meteorological conditions. A retrospective cohort study of PV-CRBSI at Tokyo Medical University Hospital (Tokyo, Japan), from 2009 to 2019, provided the data for descriptive and time series analyses used to evaluate the number of PV-CRBSI cases per 1000 admissions that occurred each month for each causative organism. By performing univariate and multivariate analyses, the researchers investigated the seasonality of cases and the relationships between meteorological conditions, other external factors, and PV-CRBSIs. This study included a total of 184 PV-CRBSI cases. The mean numbers of PV-CRBSI cases per 1000 admissions caused by all organisms, Bacillus cereus, Gram-positive cocci, and Gram-negative rods were 0.67, 0.15, 0.37, and 0.16 per month, respectively, during the study period. The time series analysis showed that the incidences of PV-CRBSI cases associated with B. cereus and Gram-negative rods were significantly different in the winter/spring from those in the summer/autumn (P < 0.05). The incidence of PV-CRBSI cases caused by B. cereus peaked during summer. The incidence of PV-CRBSI cases caused by B. cereus was significantly positively associated with average monthly temperature, whereas the incidence of PV-CRBSIs caused by Gram-negative rods was significantly negatively associated with average daylight hours. The incidence of PV-CRBSIs caused by B. cereus showed seasonality, peaking during the summer, and a significant correlation was found between PV-CRBSIs caused by B. cereus and average monthly temperature.


RNAi-mediated knockdown of Xist does not rescue the impaired development of female cloned mouse embryos.
In mice, one of the major epigenetic errors associated with somatic cell nuclear transfer (SCNT) is ectopic expression of Xist during the preimplantation period in both sexes. We found that this aberrant Xist expression could be impeded by deletion of Xist from the putative active X chromosome in donor cells. In male clones, it was also found that prior injection of Xist-specific siRNA could significantly improve the postimplantation development of cloned embryos as a result of a significant repression of Xist at the morula stage. In this study, we examined whether the same knockdown strategy could work as well in female SCNT-derived embryos. Embryos were reconstructed with cumulus cell nuclei and injected with Xist-specific siRNA at 6-7 h after oocyte activation. RNA FISH analysis revealed that siRNA treatment successfully repressed Xist RNA at the morula stage, as shown by the significant decrease in the number of cloud-type Xist signals in the blastomere nuclei. However, blastomeres with different sizes (from "pinpoint" to "cloud") and numbers of Xist RNA signals remained within single embryos. After implantation, the dysregulated Xist expression was normalized autonomously, as in male clones, to a state of monoallelic expression in both embryonic and extraembryonic tissues. However, at term there was no significant improvement in the survival of the siRNA-injected cloned embryos. Thus, siRNA injection was largely effective in repressing the Xist overexpression in female cloned embryos but failed to rescue them, probably because of an inability to mimic consistent monoallelic Xist expression in these embryos. This could only be achieved in female embryos by applying a gene knockout strategy rather than an siRNA approach.


Production of single cell protein from five strains of the microalga Chlorella spp. (Chlorophyta).
Single cell protein (SCP) production by five strains of Chlorella species (M109, M121, M122, M138, M150), isolated from different habitats, was studied under the influence of eight environmental factors. The ranges for biomass production and SCP were 150-260 mg/l, and 12.4-49.7% dry wt, respectively. The ranges of SCP under the influence of temperature, light, nitrate, ammonia, phosphorus, iron, carbonate, and sodium chloride were in the following respective order (% dry wt): 18.4-43.3, 20.5-42.3, 12.4-35.8, 15.7-41.8, 15.8-49.4, 17.4-49.7, 13.8-35.6, and 0.0-37.7. All the environmental factors had a positive effect on SCP of the five strains except that sodium chloride had a negative effect on SCP of M150. According to these results, both biomass and SCP production can be manipulated and optimized for commercial use by varying the environmental factors used here.


Photostability of gold nanoparticles with different shapes: the role of Ag clusters.
Anisotropic gold nanostructures prepared by the seed method in the presence of Ag ions have been used to study their photostability to low-power UV irradiation (254 nm) at room temperature. It has been observed that, whereas spheres are very stable to photoirradiation, rods and prisms suffer from photocorrosion and finally dissolve completely with the production of Au(III) ions. Interpretation of these differences is based on the presence of semiconductor-like Ag clusters, adsorbed onto rods and prisms, able to photocorrode the Au nanoparticles, which are absent in the case of Au spheres. We further show direct evidence of the presence of Ag clusters in Au nanorods by XANES. These results confirm a previous hypothesis (J. Am. Chem. Soc., 2014, 136, 1182-1185) about the major influence of very stable small Ag clusters, not only on the anisotropic formation of nanostructures but also on their photostability.


Correction of PCR-bias in quantitative DNA methylation studies by means of cubic polynomial regression.
DNA methylation profiling has become an important aspect of biomedical molecular analysis. Polymerase chain reaction (PCR) amplification of bisulphite-treated DNA is a processing step that is common to many currently used methods of quantitative methylation analysis. Preferential amplification of unmethylated alleles-known as PCR-bias-may significantly affect the accuracy of quantification. To date, no universal experimental approach has been reported to overcome the problem. This study presents an effective method of correcting biased methylation data. The procedure includes a calibration performed in parallel to the analysis of the samples under investigation. DNA samples with defined degrees of methylation are analysed. The observed deviation of the experimental results from the expected values is used for calculating a regression curve. The equation of the best-fitting curve is then used for correction of the data obtained from the samples of interest. The process can be applied irrespective of the locus interrogated and the number of sites analysed, avoiding an optimization of the amplification conditions for each individual locus.


Failure of Biologic Therapy in Psoriasis.
This study aims to describe the frequency of biologic therapy failure in psoriasis patients along with associated patient demographics and characteristics. This was a retrospective medical-record review of psoriasis patients evaluated from January 1st, 2013, through May 1st, 2018, and who failed at least once to adhere to their biologic therapy. Seventy-seven patients with psoriasis who had discontinued biologic therapy at least once were included in this study. Hypertension (58.4%), diabetes (37.7%), dyslipidemia (27.3%), and psoriatic arthritis (23.4%) were the main comorbidities observed. Adalimumab (ADA, 80.5%), ustekinumab (UST, 70.1%), and etanercept (ETA, 14.2%) were the most frequently used biologics in our cohort. The biologic with the longest mean duration of use prior to its discontinuation was UST (17.0 months), followed by ADA (15.9 months) and ETA (13.6 months). The most common reason for discontinuing biologic therapy was that said therapy was not effective, though for ETA and UST, the fact that biologic therapies are not universally covered by insurance company was found to be associated with their discontinuation, as well. There were no statistically significant associations found between biologic therapy discontinuation and age, gender, or comorbidities, which last included obesity, class I. Larger studies are warranted to identify risk factors associated with biologic therapy failure to help guide drug selection, decrease morbidity associated with such nonadherence and improve patient outcomes.


Working memory and attention are still impaired after three years in patients with stress-related exhaustion.
Cognitive impairment is one of the most pronounced symptoms reported by patients with stress-related mental health problems. Impairments related to executive function and to some extent speed and attention are therefore common in patients with stress-related burnout/exhaustion. In this paper we present a follow-up of cognitive performance in patients with stress-related exhaustion several years after they initially sought medical care. Thirty patients and 27 healthy controls, mean age 49 years (SD 6.5) and 55 years (SD 6.7) respectively, were included, all of whom had undergone baseline measurements of neuropsychological functioning. The mean follow-up time was three years. Half of the patients still reported mental health problems at follow-up and over time no major changes in cognitive performance were noted. The patients still performed significantly poorer than controls with regard to cognitive functions, mainly related to speed, attention and memory function. Long-lasting impairment of cognitive functions related to speed, attention and memory function noted in patients with stress-related exhaustion should be acknowledged and taken into consideration during treatment and when discussing a return to work. Follow-up periods longer than three years are needed to explore the persistence of the cognitive impairment.


Monetary policy framework in India.
In 2016, the monetary policy framework moved towards flexible inflation targeting and a six member Monetary Policy Committee (MPC) was constituted for setting the policy rate. With this step towards modernization of the monetary policy process, India joined the set of countries that have adopted inflation targeting as their monetary policy framework. The Consumer Price Index (CPI combined) inflation target was set by the Government of India at 4% with ± 2% tolerance band for the period from August 5, 2016 to March 31, 2021. In this backdrop, the paper reviews the evolution of monetary policy frameworks in India since the mid-1980s. It also describes the monetary policy transmission process and its limitations in terms of lags and rigidities. It highlights the importance of unconventional monetary policy measures in supplementing conventional tools especially during the easing cycle. Further, it examines the voting pattern of the MPC in India and compares this with that of various developed and emerging economies. The synchronization of cuts in the policy rate by MPCs of various countries during the global slowdown in 2019 and the COVID-19 pandemic in the early 2020s is also analysed.


Association between the prolonged use of magnesium sulfate for tocolysis and fracture risk among infants.
In 2013, the U.S. Food and Drug Administration issued a safety warning that cautioned against using magnesium sulfate (MgSO4) injections for more than 5 to 7 days to stop preterm delivery due to the bone problems subsequently observed in infants. However, the warning was mainly based on case reports, and further investigation is necessary to determine whether prolonged MgSO4 use increased infant fractures.To evaluate whether prolonged MgSO4 use for tocolysis increased the risk of subsequent fractures among infants.A retrospective population-based cohort study was conducted with a new-user study design using the National Health Insurance Database in Taiwan. We included pregnant women aged between 12 and 55 years old who delivered a live-born singleton. The enrollment period was from January 1, 2012 to December 31, 2014. The exposure group was defined as pregnant women who received MgSO4 injection for >5 days during pregnancy, while those not receiving any tocolytics were the reference group. The outcome was any bone fracture among the infants during the 2-year follow-up period. Propensity score matching and Cox proportional hazards regression models were used to estimate the hazard of fractures. We further studied the effect of MgSO4 treatment with varied dosages and durations of treatment in the sensitivity analyses.Among the 4092 pregnant women in the database, 693 (16.9%) of them were included in the exposure group. The hazard ratio of infant fractures among prolonged MgSO4 users was not significantly different from that of tocolytic nonusers in adjusted models (adjusted hazard ratio (aHR) = 1.48; 95% confidence interval (CI) = 0.59-3.71). A similar lack of significance was found in the sensitivity analyses (aHR = 1.45; 95% CI = 0.40-5.28 for larger treatment dosage; aHR = 2.52; 95% CI = 0.49-12.98 for longer treatment duration).Prolonged MgSO4 tocolysis use did not increase the risk of infant fractures. Our findings reconfirmed the safety of MgSO4 as a tocolytic treatment.


Consumer acceptability and sensory profile of cooked broccoli with mustard seeds added to improve chemoprotective properties.
Broccoli, a rich source of glucosinolates, is a commonly consumed vegetable of the Brassica family. Hydrolysis products of glucosinolates, isothiocyanates, have been associated with health benefits and contribute to the flavor of Brassica. However, boiling broccoli causes the myrosinase enzyme needed for hydrolysis to denature. In order to ensure hydrolysis, broccoli must either be mildly cooked or active sources of myrosinase, such as mustard seed powder, can be added postcooking. In this study, samples of broccoli were prepared in 6 different ways; standard boiling, standard boiling followed by the addition of mustard seeds, sous vide cooking at low temperature (70 °C) and sous vide cooking at higher temperature (100 °C) and sous vide cooking at higher temperature followed by the addition of mustard seeds at 2 different concentrations. The majority of consumers disliked the mildly cooked broccoli samples (70 °C, 12 min, sous vide) which had a hard and stringy texture. The highest mean consumer liking was for standard boiled samples (100 °C, 7 min). Addition of 1% mustard seed powder developed sensory attributes, such as pungency, burning sensation, mustard odor, and flavor. One cluster of consumers (32%) found mustard seeds to be a good complement to cooked broccoli; however, the majority disliked the mustard-derived sensory attributes. Where the mustard seeds were partially processed, doubling the addition to 2% led to only the same level of mustard and pungent flavors as 1% unprocessed seeds, and mean consumer liking remained unaltered. This suggests that optimization of the addition level of partially processed mustard seeds may be a route to enhance bioactivity of cooked broccoli without compromising consumer acceptability.


Teamwork culture and patient satisfaction in hospitals.
A growing line of research indicates a positive relationship between a healthcare organization's culture and various performance measures. In these studies, a key cultural characteristic is the emphasis placed on teamwork. None of the studies, however, have examined teamwork culture relative to patient satisfaction, which is now 1 of the most widely used performance measures for healthcare organizations. This study investigated the relationship between teamwork culture of hospitals and patient reports of their satisfaction with the care they received. The study setting was the Veterans Health Administration (VHA), Department of Veterans Affairs. The study sample consisted of 125 VHA hospitals for which independent and valid sources of data for culture and patient satisfaction were obtained. Each hospital's culture was assessed relative to 4 dimensions: teamwork, entrepreneurial, bureaucratic, and rational. Patient satisfaction data were available for both inpatient and outpatient settings. Results from multivariate regression analyses indicated a significant and positive relation between teamwork culture and patient satisfaction for inpatient care, and a significant and negative relation between bureaucratic culture and patient satisfaction for inpatient care. Additional analyses revealed an almost 1 standard deviation difference in patient satisfaction scores between hospitals in the top third and bottom third of the distribution for the teamwork culture measure. Study results suggest that hospitals and possibly other healthcare organizations should strive to develop a culture emphasizing teamwork and deemphasizing those aspects of bureaucracy that are not essential to assuring efficiency and quality care.


Construction and analysis of pseudogene-related ceRNA network in breast cancer.
Breast cancer (BC) is one of the leading causes of cancer-related deaths in women. The present study explored the potential role of pseudogenes in BC via construction and analysis of a competing endogenous RNA (ceRNA) network through a three-step process. First, we screened differentially expressed genes in nine BC datasets. Then the gene-pseudogenes pairs (nine hub genes) were selected according to the functional enrichment and correlation analysis. Second, the candidate hub genes and interacting miRNAs were used to construct the ceRNA network. Further analysis of the ceRNA network revealed a crucial ceRNA module with two genes-pseudogene pairs and two miRNAs. The in-depth analysis identified the GBP1/hsa-miR-30d-5p/GBP1P1 axis as a potential tumorigenic axis in BC patients. In the third step, the GBP1/hsa-miR-30d-5p/GBP1P1 axis expression level was assessed in 40 tumor/normal BC patients and MCF-7 cell lines. The expression of GBP1 and GBP1P1 was significantly higher in the tumor compared to the normal tissue. However, the expression of hsa-miR-30d-5p was lower in tumor samples. Then, we introduced the GBP1P1 pseudogene into the MCF-7 cell line to evaluate its effect on GBP1 and hsa-miR-30d-5p expression. As expected, the GBP1 level increased while the hsa-miR-30d-5p level decreased in the GBP1P1-overexprsssing cell line. In addition, the oncogenic properties of MCF-7 (cell viability, clonogenicity, and migration) were improved after GBP1P1 overexpression. In conclusion, we report a ceRNA network that may provide new insight into the role of pseudogenes in BC development.


Of time and the moon.
Considerable information concerning lunar chronology has been obtained by the study of rocks and soil returned by the Apollo 11 and Apollo 12 missions. It has been shown that at the time the moon, earth, and solar system were formed, approximately 4.6 approximately 10(9) years ago, a severe chemical fractionation took place, resulting in depletion of relatively volatile elements such as Rb and Pb from the sources of the lunar rocks studied. It is very likely that much of this material was lost to interplanetary space, although some of the loss may be associated with internal chemical differentiation of the moon. It has also been shown that igneous processes have enriched some regions of the moon in lithophile elements such as Rb, U, and Ba, very early in lunar history, within 100 million years of its formation. Subsequent igneous and metamorphic activity occurred over a long period of time; mare volcanism of the Apollo 11 and Apollo 12 sites occurred at distinctly different times, 3.6 approximately 10(9) and 3.3 approximately 10(9) years ago, respectively. Consequently, lunar magmatism and remanent magnetism cannot be explained in terms of a unique event, such as a close approach to the earth at a time of lunar capture. It is likely that these phenomena will require explanation in terms of internal lunar processes, operative to a considerable depth in the moon, over a long period of time. These data, together with the low present internal temperatures of the moon, inferred from measurements of lunar electrical conductivity, impose severe constraints on acceptable thermal histories of the moon. Progress is being made toward understanding lunar surface properties by use of the effects of particle bombardment of the lunar surface (solar wind, solar flare particles, galactic cosmic rays). It has been shown that the rate of micrometeorite erosion is very low (angstroms per year) and that lunar rocks and soil have been within approximately a meter of the lunar surface for hundreds of millions of years. Future work will require sampling distinctly different regions of the moon in order to provide data concerning other important lunar events, such as the time of formation of the highland regions and of the mare basins, and of the extent to which lunar volcanism has persisted subsequent to the first third of lunar history. This work will require a sufficient number of Apollo landings, and any further cancellation of Apollo missions will jeopardize this unique opportunity to study the development of a planetary body from its beginning. Such a study is fundamental to our understanding of the earth and other planets.


Direct and Mediated Effects of a Social-Emotional and Character Development Program on Adolescent Substance Use.
Mitigating and preventing substance use among adolescents requires approaches that address the multitude of factors that influence this behavior. Such approaches must be tested, not only for evidence of empirical effectiveness, but also to determine the mechanisms by which they are successful. The aims of the present study were twofold: 1) To determine the effectiveness of a school-based social-emotional and character development (SECD) program, Positive Action (PA), in reducing substance use (SU) among a sample of U.S. youth living in a low-income, urban environment, and 2) to test one mechanism by which the program achieves its success. We used longitudinal mediation analysis to test the hypotheses that: 1) students attending PA intervention schools engage in significantly less SU than students attending control schools, 2) students attending PA intervention schools show significantly better change in SECD than students attending control schools, and 3) the effect of the PA intervention on SU is mediated by the change in SECD. Analyses revealed program effects on both SECD and SU, a relationship between SECD and SU, and the effects of PA on SU were completely mediated by changes in SECD. Future research directions and implications for school-based social-emotional and character development efforts and substance use prevention are addressed.


The acceptability of suicide among rural residents, urban residents, and college students from three locations in China: a cross-sectional survey.
Community attitudes about suicide and their relationship to suicidal behavior have not been adequately investigated in low- and middle-income countries. To compare the acceptability of suicide in different population cohorts in China, identify factors that affect the degree of acceptability, and assess the relationship of cohort-specific acceptability of suicide and suicide rates. A multistage stratified random sample of 608 rural residents, 582 urban residents, and 629 college students were administered a 25-item scale studying the likelihood they would consider suicide (on a 5-point Likert scale) if they experienced a variety of stressors ranging from "being disciplined at work" to "developing a chronic mental illness." The internal consistency and test-retest reliability for the scale are excellent (Cronbach's alpha = 0.92, ICC = 0.75). College students had the most permissive attitudes about suicide, and urban residents were significantly more accepting of suicide as a response to serious life stressors than were rural residents. Multivariate analysis found that the overall acceptability score was higher in women, decreased with age, and increased with years of education. There was no clear relationship between cohort-specific acceptance of suicide and reported rates of suicide, highlighting the complexity of the relationship between attitudes about suicide (of which acceptability is only one component) and suicidal behavior.


Association between Clostridium difficile infection and antimicrobial usage in a large group of English hospitals.
This study aimed to determine the association between the reduction in the number of Clostridium difficile infection (CDI) cases reported by the English National Health Service (NHS) hospitals and concurrent antimicrobial use. A retrospective ecological study for January 2005 to December 2008 was conducted using data from 26 of the 29 NHS trusts (i.e. a trust manages one or more hospitals) located in the North West Strategic Health Authority of England. Antimicrobial use data, for patients of all ages, were provided by IMS Health, and CDI case data for patients aged ≥65 years were provided by the Health Protection Agency. Antimicrobial use was converted into defined daily doses (DDDs). The overall association between antimicrobial use and CDI for the trusts was investigated using multilevel models. Our study shows a positive significant association between the CDI cases and the use of the following antimicrobials: 'third-generation cephalosporins' [11.62 CDI cases per 1000 DDDs; 95% confidence interval (CI), 5.92–17.31]; 'fluoroquinolones' (4.79 CDI cases per 1000 DDDs; 95% CI, 2.83–6.74); and 'second-generation cephalosporins' (4.25 CDI cases per 1000 DDDs; 95% CI, 1.66–6.83). The strength of this association was not significantly different (95% CI) among the antimicrobial groups. This study shows that the reduction in the number of CDI cases reported by the English NHS hospitals is associated with concurrent reductions in antimicrobial use. This means that the number of CDI cases over time decreased in a similar fashion to the usage of various antimicrobials.


Red cell fragmentation in human disease (a light and scanning electron microscope study).
Although the mechanism of schizocyte formation has been amply documented in animal experiments and in in-vitro models, the fragmentation encounter between flowing red cells and fibrin strands has not previously been successfully demonstrated in human, microangiopathic disease. If blood flow is restored briefly in vitro immediately prior to tissue fixation, the instant of red cell fragmentation can be captured. Examination of tissue specimens fixed in this manner shows a pathophysiologic process that amplifies the findings previously described in other studies. In the patient under study, the microangiopathic process was widespread in all specimens of pulmonary and renal tissue that had been fixed after brief restoration of blood flow. Small arteries as well as the true microcirculation of both organs were involved. The microangiopathic process in the small arteries and arterioles presented as a partially occlusive thrombus of characteristic histology. The pulmonary capillaries contained linear fibrin microclots festooned with distorted and partially fragmented red cells. The microcirculation of the kidney showed the same findings as well as amorphous, sludged, occlusive red cell masses, particularly in the renal medulla.


Expression of CP4 EPSPS in microspores and tapetum cells of cotton (Gossypium hirsutum) is critical for male reproductive development in response to late-stage glyphosate applications.
Plants expressing Agrobacterium sp. strain CP4 5-enolpyruvylshikimate-3-phosphate synthase (CP4 EPSPS) are known to be resistant to glyphosate, a potent herbicide that inhibits the activity of the endogenous plant EPSPS. The RR1445 transgenic cotton line (current commercial line for Roundup Ready Cotton) was generated using the figwort mosaic virus (FMV) 35S promoter to drive the expression of the CP4 EPSPS gene, and has excellent vegetative tolerance to glyphosate. However, with high glyphosate application rates at developmental stages later than the four-leaf stage (late-stage applications: applications that are inconsistent with the Roundup labels), RR1445 shows male sterility. Another transgenic cotton line, RR60, was generated using the FMV 35S promoter and the Arabidopsis elongation factor-1alpha promoter (AtEF1alpha) for the expression of CP4 EPSPS. RR60 has excellent vegetative and reproductive tolerance to applications of glyphosate at all developmental stages. Histochemical analyses were conducted to examine the male reproductive development at the cellular level of these cotton lines in response to glyphosate applications, and to investigate the correlation between glyphosate injury and the expression of CP4 EPSPS in male reproductive tissues. The expression of CP4 EPSPS in RR60 was found to be strong in all male reproductive cell types. Conversely, CP4 EPSPS expression in RR1445 was low in pollen mother cells, male gametophytes and tapetum, three crucial male reproductive cell types. Our results indicate that the FMV 35S promoter, although expressing strongly in most vegetative tissues in plants, has extremely low activity in these cell types.


The bacterial promoter spacer modulates promoter strength and timing by length, TG-motifs and DNA supercoiling sensitivity.
Transcription, the first step to gene expression, is a central coordination process in all living matter. Besides a plethora of regulatory mechanisms, the promoter architecture sets the foundation of expression strength, timing and the potential for further regulatory modulation. In this study, we investigate the effects of promoter spacer length and sequence composition on strength and supercoiling sensitivity in bacteria. Combining transcriptomics data analysis and standardized synthetic promoter libraries, we exclude effects of specific promoter sequence contexts. Analysis of promoter activity shows a strong variance with spacer length and spacer sequence composition. A detailed study of the spacer sequence composition under selective conditions reveals an extension to the -10 region that enhances RNAP binding but damps promoter activity. Using physiological changes in DNA supercoiling levels, we link promoter supercoiling sensitivity to overall spacer GC-content. Time-resolved promoter activity screens, only possible with a novel mild treatment approach, reveal strong promoter timing potentials solely based on DNA supercoiling sensitivity in the absence of regulatory sites or alternative sigma factors.


Perioperative Glycemic Management in Cardiac Surgery: A Narrative Review.
Diabetes and hyperglycemic events in cardiac surgical patients are associated with postoperative morbidity and mortality. The causes of dysglycemia, the abnormal fluctuations in blood glucose concentrations, in the perioperative period include surgical stress, surgical techniques, medications administered perioperatively, and patient factors. Both hyperglycemia and hypoglycemia lead to poor outcomes after cardiac surgery. While trying to control blood glucose concentration tightly for better postoperative outcomes, hypoglycemia is the main adverse event. Currently, there is no definite consensus on the optimum perioperative blood glucose concentration to be maintained in cardiac surgical patients. This review provides an overview of perioperative glucose homeostasis, the pathophysiology of dysglycemia, factors that affect glycemic control in cardiac surgery, and current practices for glycemic control in cardiac surgery.


Flavonoids reduces lipopolysaccharide-induced release of inflammatory mediators in human bronchial epithelial cells: Structure-activity relationship.
Flavonoids are polyphenolic compounds that are widely present in food and Chinese medicine. The aim of the present study was to identify the flavonoids with anti-inflammatory effects in the airway; and to determine the role of anti-oxidant and cyclic adenosine monophosphate (cAMP) in the anti-inflammatory effect. Human bronchial epithelial BEAS-2B cells were exposed to bacterial endotoxin lipopolysaccharide (LPS) in the absence or presence of different flavonoids, which are categorized according to their chemical structures in seven subclasses [anthocyanidins, chalcones, flavanes, flavanones, flavones, flavonols, isoflavones]. Among the 17 flavonoids tested, only apigenin (flavones), luteolin (flavones), daidzein (isoflavones) and genistein (isoflavones) reduced LPS-induced release of inflammatory cytokines/chemokines interleukin (IL)-6, IL-8 and monocyte chemoattractant protein-1 in BEAS-2B cells. Quercetin caused further increase in LPS-induced IL-6 and IL-8 levels. It alone significantly increased nuclear factor-kappa B (NF-κB) p65 activity and the cellular oxidative stress marker malondialdehyde (MDA) level in BEAS-2B cells. By contrast, apigenin and genistein reduced LPS-induced increases in nuclear NF-κB activity and MDA level. Apigenin and genistein, but not quercetin, increased the cAMP level in BEAS-2B cells, and the cell-permeable cAMP analogue, 8-Br-cAMP, inhibited LPS-induced increase of IL-8 level. These findings suggest that the presence of C5-OH, C7-OH, C2=C3 and C4=O functional groups in the flavonoids is associated with greater anti-inflammatory effect, while that of C3-OH or glycosylation group at the A-ring greatly decreased the anti-inflammatory effect. The anti-inflammatory effect of these flavonoids may be related to their anti-oxidant properties, and partly to their ability in increasing cAMP level.


Reliability and validity of the public transportation use assessment form for individuals after stroke.
To examine the reliability and validity of the public transportation use assessment form (PTAF), which was developed for assessing the performance of tasks during public transportation use. Fifty consecutive patients admitted after a stroke to a convalescent rehabilitation hospital and received field-based practice for public transportation use were enrolled. A physical therapist (PT) and an occupational therapist (OT) independently evaluated actual participant performance using the PTAF. Its internal consistency, inter-rater reliability, and construct validity were examined against other clinical measures related to the ability to use public transportation. Cronbach's coefficient alpha for the internal consistency for overall PTAF was 0.84 and 0.88 for PTs and OTs, respectively. Cohen's weighted κ coefficient for the inter-rater reliability for each item ranged from 0.61 to 0.83. Intraclass correlation coefficients for the inter-rater reliability for the mean scores of the items comprising the PTAF were 0.90 for all 14 items, 0.76 for items required for train use only, and 0.88 for items required for bus use only. The correlation coefficients for the construct validity between PTAF and clinical measures ranged from 0.38 to 0.59 (p < 0.05). The PTAF showed sufficient internal consistency, intra-rater reliability, and construct validity.Implications for rehabilitationThis study illustrated the inter-rater reliability of the public transportation use assessment form (PTAF), indicating that the PTAF can be used for reliable assessment independent of the rater.The PTAF showed good internal consistency, indicating that each item in the PTAF consistently assessed the ability of patients with stroke to use public transportation.The PTAF correlated with assessment tools such as walking ability, balance, motor paralysis, intelligence, and activities of daily living, indicating that it reflects the functions and abilities necessary to use public transportation.


Neutrophil extracellular traps mediate articular cartilage damage and enhance cartilage component immunogenicity in rheumatoid arthritis.
Rheumatoid arthritis (RA) is characterized by synovial joint inflammation, cartilage damage, and dysregulation of the adaptive immune system. While neutrophil extracellular traps (NETs) have been proposed to play a role in the generation of modified autoantigens and in the activation of synovial fibroblasts, it remains unknown whether NETs are directly involved in cartilage damage. Here, we report a new mechanism by which NET-derived elastase disrupts cartilage matrix and induces release of membrane-bound peptidylarginine deiminase-2 by fibroblast-like synoviocytes (FLSs). Cartilage fragments are subsequently citrullinated, internalized by FLSs, and then presented to antigen-specific CD4+ T cells. Furthermore, immune complexes containing citrullinated cartilage components can activate macrophages to release proinflammatory cytokines. HLA-DRB1*04:01 transgenic mice immunized with NETs develop autoantibodies against citrullinated cartilage proteins and display enhanced cartilage damage. Inhibition of NET-derived elastase rescues NET-mediated cartilage damage. These results show that NETs and neutrophil elastase externalized in these structures play fundamental pathogenic roles in promoting cartilage damage and synovial inflammation. Strategies targeting neutrophil elastase and NETs could have a therapeutic role in RA and in other inflammatory diseases associated with inflammatory joint damage.