Cellular toxicities of gadolinium-based contrast agents used in magnetic resonance imaging.
Contrast agents have been used in magnetic resonance imaging (MRI) as a radiological method. Gadolinium-based contrast agents (GBCAs), because of their paramagnetic characteristics, are the ones mostly used in MRI to increase signal intensity. However, the use of contrast media has raised concerns on cellular toxic risks of these agents. Studies showed the accumulation of gadolinium after injection to humans with or without renal impairment. Also, there are findings obtained under in vitro and/or in vivo conditions that revealed conflicting results for their cytotoxic and genotoxic effects. Some of them declared damage in cells and genetic material; some others did not. Abnormal cell growth and genetic aberration are critical because they may lead to carcinogenesis in somatic cells or may be transferred to the next generations through germ cells. Therefore, understanding the effect of GBCAs on cells is important for their safer usage in clinical administrations to generate high-quality contrast-enhanced magnetic resonance images. Because of all these reasons, cellular toxicities-mainly genotoxic and cytotoxic effects-of GBCAs were reviewed in this paper.


Luminal and intracellular cGMP inhibit the mTAL reabsorptive capacity through different pathways.
Since, in the presence of ANF, urinary cGMP was shown to be of glomerular origin, a possible paracrine effect of luminal cGMP on the medullary thick ascending limb (mTAL) function was investigated. Net chloride reabsorption (JCl) was determined on isolated microperfused tubules from mouse kidney. Addition of 10(-6) M cGMP to the lumen significantly decreased JCl by 46.5 +/- 4.6%. A concentration-dependent decrease of the transepithelial voltage was observed, with a 10(-8) M threshold. Added to the bath, ANF (10(-7) M) as well as urodilatin (6 x 10(-8) M) decreased JCl by 29.8 +/- 3.9% and 36.9 +/- 5.1%, respectively, an effect reproduced by 8-bromo cGMP and associated with a significant increase in tubular cGMP content. The inhibitory effect of ANF was similar whether or not cGMP was present in the lumen. Furthermore, increasing intracellular cGMP content by 8-bromo cGMP did not prevent a further effect of luminal cGMP. Finally, H-8, which blocked the effect of ANF, urodilatin, and 8-bromo cGMP, failed to abolish the luminal cGMP-induced decrease of JCl, suggesting that this effect did not require a cGMP-dependent protein kinase activation. It is concluded that luminal cGMP inhibits the reabsorptive function of the mTAL through a pathway different from the intracellular cGMP production.


Acute-on-chronic liver failure: Definitions, pathophysiology and principles of treatment.
The term acute-on-chronic liver failure (ACLF) defines an abrupt and life-threatening worsening of clinical conditions in patients with cirrhosis or chronic liver disease. In recent years, different definitions and diagnostic criteria for the syndrome have been proposed by the major international scientific societies. The main controversies relate to the type of acute insult (specifically hepatic or also extrahepatic), the stage of underlying liver disease (cirrhosis or chronic hepatitis) and the concomitant extrahepatic organ failure(s) that should be considered in the definition of ACLF. Therefore, different severity criteria and prognostic scores have been proposed and validated. Current evidence shows that the pathophysiology of ACLF is closely associated with an intense systemic inflammation sustained by circulating pathogen-associated molecular patterns and damage-associated molecular patterns. The development of organ failures may be a result of a combination of tissue hypoperfusion, direct immune-mediated damage and mitochondrial dysfunction. Management of ACLF is currently based on the supportive treatment of organ failures, mainly in an intensive care setting. For selected patients, liver transplantation is an effective treatment that offers a good long-term prognosis. Future studies on potential mechanistic treatments that improve patient survival are eagerly awaited.


Characteristics associated with the willingness to receive a COVID-19 vaccine and an exploration of the general public's perceptions: A mixed-methods approach.
Demographics and media discourse impact vaccine hesitancy. We explored the New Zealand public's perceptions of COVID-19 vaccines and associated media portrayal, and determined predictive factors associated with willingness to receive vaccines. A community cohort (N = 340) completed online surveys. A logistic regression explored whether characteristics predict willingness to receive the vaccine. Textual data were analysed thematically. Willingness to receive the vaccine was high (90%). Having a postgraduate degree (p =.026), trying to receive an influenza vaccine (p <.001) and fewer concerns (p <.001) predicted willingness. Health keyworkers (p <.001) were less willing. Participants wanted the vaccine for protection and returning to normality. Reasons against receiving vaccines regarded safety, efficacy, and an unclear roll-out plan. The media was reported to generally provide good/positive coverage, but also engage in unbalanced reporting and spreading misinformation. Education strategies should include collaborations between media and scientists and focus on distributing easy-to-access information. Health keyworkers should be reassured of testing/safety.


Skp2 expression unfavorably impacts survival in resectable esophageal squamous cell carcinoma.
The correlation of S-phase kinase-associated protein 2 (Skp2) with metastasis and prognosis in esophageal squamous cell carcinoma (ESCC) is controversial. The purpose of this study was to explore whether there was a correlation between the expression of Skp2 evaluated by immunohistochemistry and the clinical outcome of patients with operable ESCC, and to further determine the possible mechanism of the impact of Skp2 on survival. Tissue microarrays that included 157 surgically resected ESCC specimens was successfully generated for immunohistochemical evaluation. The clinical/prognostic significance of Skp2 expression was analyzed. Kaplan-Meier analysis was used to compare the postoperative survival between groups. The prognostic impact of clinicopathologic variables and Skp2 expression was evaluated using a Cox proportional hazards model. A cell proliferation assay and a colony formation assay were performed in ESCC cell lines to determine the function of Skp2 on the progression of ESCC in vitro. Skp2 expression correlated closely with the T category (p = 0.035) and the pathological tumor-node-metastasis (TNM) stage (p = 0.027). High expression of Skp2 was associated with poor overall survival in resectable ESCC (p = 0.01). The multivariate Cox regression analysis demonstrated that pathological T category, pathological N category, cell differentiation, and negative Skp2 expression were independent factors for better overall survival. In vitro assays of ESCC cell lines demonstrated that Skp2 promoted the proliferative and colony-forming capacity of ESCCs. Negative Skp2 expression in primary resected ESCC is an independent factor for better survival. Skp2 may play a pro-proliferative role in ESCC cells.


Validation of the MG-DIS: a disability assessment for myasthenia gravis.
This paper is aimed to present the validation of the myasthenia gravis disability assessment (MG-DIS), a MG-specific patient-reported disability outcome measure. Consecutive MG patients were enrolled, followed-up for 12 months and administered the SF-36, the WHO disability assessment schedule (WHODAS 2.0) and the preliminary 31-item MG-DIS addressing impairments and activity limitations. Factor structure and metric properties were assessed. In total, 109 patients were enrolled: 76 were females, mean age 50, mean MG duration 10.4 years, 86 were AChR-positive. The MG-DIS was reduced to 20 items, explaining 70.6 % of the original questionnaire variance, four subscales (generalized impairment-related problems; bulbar function-related problems; mental health and fatigue-related problems; vision-related problems) and an overall disability index. The MG-DIS has good metric properties (Cronbach's alpha ranging between .808 and .930), is stable, showed to be more sensitive than the WHODAS 2.0 and SF-36 to detect group differences and longitudinal changes and was well correlated with the MG-composite (.642). The MG-DIS includes items representing ocular, generalized, bulbar and respiratory symptoms, and is therefore well-built around MG-specific features. MG-DIS can be used in clinical trials as well as in observational or epidemiological studies to characterize patients' disability level and address the amount of improvement in disability. Further studies are needed to explore the possibility of a shorter disability scale.


Fungal infection has sublethal effects in a lowland subtropical amphibian population.
The amphibian chytrid fungus, Batrachochytrium dendrobatidis (Bd), has been implicated as a primary cause of decline in many species around the globe. However, there are some species and populations that are known to become infected in the wild, yet declines have not been observed. Here we conducted a yearlong capture-mark-recapture study and a 2-year long disease monitoring study of northern cricket frogs, Acris crepitans, in the lowland subtropical forests of Louisiana. We found little evidence for an impact of Bd infection on survival; however, Bd infection did appear to cause sublethal effects, including increased capture probability in the field. Our study suggests that even in apparently stable populations, where Bd does not appear to cause mortality, there may be sublethal effects of infection that can impact a host population's dynamics and structure. Understanding and documenting such sublethal effects of infection on wild, seemingly stable populations is important, particularly for predicting future population declines.


The use of laryngeal mucosa mesenchymal stem cells for the repair the vocal fold injury.
Stem cell transplantation is a kind of attractive and new approach that complements traditional restorative or surgical techniques for the regeneration of injured or pathologically damaged laryngeal tissues. However, the best cell delivery strategy remains to be identified. The objective of this study was to establish a new strategy to the healing of injured vocal fold, using laryngeal mucosa mesenchymal stem cells differentiating into myofibroblasts or fibroblasts and improving the reconstruction microenvironment in the vocal fold injury as a new alternative as seed cells for laryngeal tissue engineering. After isolation and expansion, cells were identified as adherent mesenchymal cells with substantial proliferation potential in vitro, and were also characterized by flow cytometry. The differentiation potential of mesenchymal cells was maintained during proliferation as confirmed by culturing for adipogenesis, osteogenesis and chondrocyte. When LM-MSC was transplanted into the injured vocal fold, it has the potent differentiated into myofibroblasts and fibroblasts, which could regulate extracellular matrix, block collagen and the fibronectin rapid increased, inhibit the rapidly decrease of elastic fiber and HA, decrease the microenvironment inflammatory reaction, and prevent the formation of vocal fold scar.


Coherence effects in scattering systems of two interacting dipoles at resonance.
Coherence effects in the backward-scattering direction, such as enhanced backscattering (EBS) and the appearance of a negative polarization branch (NPB), are investigated for a simple scattering system composed of two dipolelike particles separated by a fixed distance. The influence on these effects of coupling resonances between the particles is analyzed. The appearance of these coherent phenomena (EBS and NPB) and their relationship as a function of the optical properties of the particles (polarizability) and particle separation are also studied.


The role of operative intervention in management of congenital extrahepatic portosystemic shunt.
Congenital extrahepatic portosystemic shunt (CEPS) is a rare venous malformation in which mesenteric venous blood drains directly into the systemic circulation. It is still a matter of debate whether conservative or operative strategies should be used to treat symptomatic CEPS. The aim of this study was to evaluate the role of operative intervention in the management of CEPS. Between June 2004 and August 2010, 6 consecutive patients with symptomatic CEPS were treated in our department. There were 3 male and 3 female patients, with a median age of 3.5 years (range, 1-8). Their demographic, clinical, and laboratory data were analyzed. All patients were scheduled to undergo shunt ligation or liver transplantation (LT). Living donor LT was carried out in 4 patients, and shunt ligation in 2. After a median follow-up of 25 months, all the patients are alive currently with marked relief of symptoms. Shunt ligation or LT for symptomatic CEPS is potentially curative.


The expression of scavenger receptor class B, type I (SR-BI) and caveolin-1 in parenchymal and nonparenchymal liver cells.
The liver is the major site of cholesterol synthesis and metabolism, and the only substantive route for eliminating blood cholesterol. Scavenger receptor class B, type I (SR-BI) has been reported to be responsible for mediating the selective uptake of high-density lipoprotein cholesteryl esters (HDL-CE) in liver parenchymal cells (PC). We analysed the expression of SR-BI in isolated rat liver cells, and found the receptor to be highly expressed in liver PC at both the mRNA and protein levels. We also found SR-BI to be expressed in liver endothelial cells (LEC) and Kupffer cells (KC). SR-BI has not previously been reported to be present in LEC. CD36 mRNA was expressed in all three liver cell types. Since caveolin-1 appears to colocalize with SR-BI and CD36 in caveolae of several cell lines, the distribution and expression of caveolin-1 in the liver cells were investigated. Caveolin-1 was not detected in PC but was found in both LEC and KC. This led to the suggestion that caveolin-1 may be more important in the efflux of cholesterol than in the selective uptake of cholesterol in the liver.


Environmental and economic aspects of water kiosks: case study of a medium-sized Italian town.
The consumption of bottled water in Italy began in the 1970s. Since then, this usage has grown considerably, also as a result of changes in habits. The environmental impact as a result of the water production chain is very significant; it would be considered, for example, the use of plastic bottles, the consumption of oil in the production of the bottles, the emission of air from the vehicles that transport the bottles, non-recycled plastic packaging, etc. In this study, considering the comparison between two situations, use of bottled water and use of water kiosk (WK), an environmental and economic impact evaluation has been done. The study considered the production of a WK in a town with 9000 inhabitants, which supplies controlled, still and sparkling water, with an organoleptic quality higher than tap water coming from the aqueduct. In particular, taking into consideration the environmental aspects, specific attention was paid both to CO2 emissions and PET bottle waste reduction. The economic impact evaluation was carried out from the consumer's point of view. In order to provide a supply service that was economically sustainable, a calculation was done with the aim of determining a specific fee for the supplied water. Moreover, a comparison has been made between quality parameters achieved with the analysis of water from aqueducts with the limits established in the Italian legislation and the parameters of several Italian water brands. The study has the aim at considering the opportunity to follow a different people's habits, closer to the concept of sustainability, reducing the environmental charge related to the realization, transport and consumption of plastic water bottles without significant reduction of the quality of the service and with convenient and interesting economic implications. In fact the results of the study show that the alternative of WKs is more efficient in economic and environmental terms respect to the use of bottled water.


Chiral and isomeric analysis by electrospray ionization and sonic spray ionization using the fixed-ligand kinetic method.
The fixed-ligand version of the kinetic method has been used for chiral and for isomeric analysis by studying the dissociation kinetics of transition metal-bound trimeric cluster ions ([(M(II) + L(fixed)-H)(ref*)(An)](+), where M(II) is a transition metal, L fixed is a fixed (non-dissociating) ligand, ref* is a reference ligand and An is the analyte. The trimeric cluster ions are readily generated by electrospray ionization (ESI) or sonic spray ionization (SSI). The size of the fixed ligand, L- Phe-Gly-L-P he-Gly, is chosen based on previous results but with the inclusion of aromatic functionality to increase chiral recognition. Improved chiral/isomeric differentiation results from enhanced chiral/isomeric interactions (metal-ligand and ligand-ligand) due to the fixed ligand. As shown in the cases of chiral dipeptides (D-Ala-D-Ala/L-Ala-L-Ala), sugars (D/L-glucose, D/L-mannose) and isomeric tetrapeptides (L-Ala-Gly-Gly-Gly/Gly-Gly -Gly-L-Ala), improved chiral/isomeric discrimination by factors from three to six were obtained by the fixed ligand procedure. Chiral recognition is independent of the concentrations of the analyte, the reference ligand, the fixed ligand and the transition metal salt, a great advantage for practical applications. In addition to increased chiral distinction, the simplified dissociation kinetics also contribute to improved accuracy in chiral quantification, in comparison with data obtained by investigating the dissociation kinetics of simple trimeric cluster ions [M(II)(ref*)2(An) H](+). Accurate determination of enantiomeric excess (ee) is demonstrated by enantiomeric quantification of D-Ala-D-Ala/L-Ala-L-Ala down to 2% ee. Both ESI and SSI allow chiral quantification with similar accuracies. The performance of chiral analysis experiments is not limited to ion trapping devices such as quadrupole ion trap mass spectrometers by a hybrid quadrupole-time of flight (Q-ToF) mass spectrometer is shown to provide an alternative choice. The fixed-ligand kinetic method is not restricted to any particular kinds of isomers and, hence, represents a general procedure for improving molecular recognition and chiral analysis in the gas phase.


The Wisdom in Virtue: Pursuit of Virtue Predicts Wise Reasoning About Personal Conflicts.
Most people can reason relatively wisely about others' social conflicts, but often struggle to do so about their own (i.e., Solomon's paradox). We suggest that true wisdom should involve the ability to reason wisely about both others' and one's own social conflicts, and we investigated the pursuit of virtue as a construct that predicts this broader capacity for wisdom. Results across two studies support prior findings regarding Solomon's paradox: Participants ( N = 623) more strongly endorsed wise-reasoning strategies (e.g., intellectual humility, adopting an outsider's perspective) for resolving other people's social conflicts than for resolving their own. The pursuit of virtue (e.g., pursuing personal ideals and contributing to other people) moderated this effect of conflict type. In both studies, greater endorsement of the pursuit of virtue was associated with greater endorsement of wise-reasoning strategies for one's own personal conflicts; as a result, participants who highly endorsed the pursuit of virtue endorsed wise-reasoning strategies at similar levels for resolving their own social conflicts and resolving other people's social conflicts. Implications of these results and underlying mechanisms are explored and discussed.


Integrating New Learning Methods into Equine Nutrition Classrooms: The Importance of Students' Perceptions.
Despite greater efforts made over recent years to teach animal nutrition to veterinary students, more needs to be done to develop teaching methods and improve education on equine nutrition. The flipped classroom and peer-assisted learning (FC/PAL) approach is a teaching technique suggested to support learning; however, research into FC/PAL in veterinary education is scarce and little is known about students' views on this approach. This study examined the results of a questionnaire administered to assess veterinary students' perceptions of/views about the suitability of FC/PAL applied in an equine nutrition course. Correlation analysis of students' responses to questionnaire items and principal component analysis (PCA) was performed to analyze the variability in student perceptions. The results showed that although half the students reported to be uninterested in equine nutrition, most expressed positive attitudes towards FC/PAL. Comparing the perceptions/views towards FC/PAL among low and high achievers, a significant difference was found for the item considering the duration of FC/PAL (P = .002). Correlation analysis between questionnaire items showed a predominance of positive moderate correlations among high achievers and weak correlations among low achievers. PCA revealed three important findings. First, student interest towards equine nutrition explains the response variability among low achievers. Second, there was significant agreement among almost all high achievers regarding the duration of the FC/PAL. Third, in the item evaluating the usefulness of collaborating with peers, disagreement was only found among low achievers. In summary, a flipped classroom combined with the same level peer-assisted learning appears to encourage student interaction and collaboration in the classroom.


Emergence of the East-Central-South-African genotype of Chikungunya virus in Brazil and the city of Rio de Janeiro may have occurred years before surveillance detection.
Brazil, which is hyperendemic for dengue virus (DENV), has had recent Zika (ZIKV) and (CHIKV) Chikungunya virus outbreaks. Since March 2016, CHIKV is the arbovirus infection most frequently diagnosed in Rio de Janeiro. In the analysis of 1835 syndromic patients, screened by real time RT-PCR, 56.4% of the cases were attributed to CHIKV, 29.6% to ZIKV, and 14.1% to DENV-4. Sequence analyses of CHIKV from sixteen samples revealed that the East-Central-South-African (ECSA) genotype of CHIKV has been circulating in Brazil since 2013 [95% bayesian credible interval (BCI): 03/2012-10/2013], almost a year before it was detected by arbovirus surveillance program. Brazilian cases are related to Central African Republic sequences from 1980's. To the best of our knowledge, given the available sequence published here and elsewhere, the ECSA genotype was likely introduced to Rio de Janeiro early on 2014 (02/2014; BCI: 07/2013-08/2014) through a single event, after primary circulation in the Bahia state at the Northestern Brazil in the previous year. The observation that the ECSA genotype of CHIKV was circulating undetected underscores the need for improvements in molecular methods for viral surveillance.


Is there tachyphylaxis to intravitreal anti-vascular endothelial growth factor pharmacotherapy in age-related macular degeneration?
To determine whether repetitive injections of intravitreal bevacizumab and/or triamcinolone acetate in patients with exudative age-related macular degeneration (AMD) results in a decrease in biological response. Retrospective comparative case series. Forty-three eyes of 43 patients with exudative AMD. Pre- and postinjection optical coherence tomography (OCT) sections of 43 patients with AMD were analyzed to determine the change in the biologic response after each subsequent injection of intravitreal bevacizumab (2.5 mg/100 microL), preservative-free triamcinolone acetonide (pfTA) (4.0 mg/100 microL), or a combination of bevacizumab (1.25 mg/50 microL) and pfTA (2.0 mg/50 microL). The retinal thickness of each OCT sector was determined and expressed as volume. Standardized volumetric change index (SVCI) was determined to identify a statistically significant change. Pre- and postinjection (6 weeks) SVCI differences were plotted as a function of time to determine the biological response after each intravitreal treatment. Change in SVCI after intravitreal injections and the number of injections required to decrease the biological response by 50% (INJ(50)). There was no difference in the age, gender, and preinjection thickness of the retina in each of the 3 groups. The SVCI after intravitreal bevacizumab injections decreased, indicating a possible tachyphylactic response to bevacizumab. This decrease in biological response was partially alleviated with the addition of pfTA. Combination of pfTA and bevacizumab increased the INJ(50) from 2.9 with bevacizumab alone to 5.1 injections. A biphasic biologic response was observed with pfTA characterized by a rapid increase in efficacy with the second injection, peaking at the third injection and gradually decreasing afterward. Repeated intravitreal injections of bevacizumab in exudative AMD seemed to be associated with decreased bioefficacy. However, combined pharmacotherapy with triamcinolone acetate lessened this effect. Thus, multitargeted pharmacotherapy in exudative AMD may have a therapeutic benefit. Proprietary or commercial disclosure may be found after the references.


Health and economic impact of air pollution in the states of India: the Global Burden of Disease Study 2019.
The association of air pollution with multiple adverse health outcomes is becoming well established, but its negative economic impact is less well appreciated. It is important to elucidate this impact for the states of India. We estimated exposure to ambient particulate matter pollution, household air pollution, and ambient ozone pollution, and their attributable deaths and disability-adjusted life-years in every state of India as part of the Global Burden of Disease Study (GBD) 2019. We estimated the economic impact of air pollution as the cost of lost output due to premature deaths and morbidity attributable to air pollution for every state of India, using the cost-of-illness method. 1·67 million (95% uncertainty interval 1·42-1·92) deaths were attributable to air pollution in India in 2019, accounting for 17·8% (15·8-19·5) of the total deaths in the country. The majority of these deaths were from ambient particulate matter pollution (0·98 million [0·77-1·19]) and household air pollution (0·61 million [0·39-0·86]). The death rate due to household air pollution decreased by 64·2% (52·2-74·2) from 1990 to 2019, while that due to ambient particulate matter pollution increased by 115·3% (28·3-344·4) and that due to ambient ozone pollution increased by 139·2% (96·5-195·8). Lost output from premature deaths and morbidity attributable to air pollution accounted for economic losses of US$28·8 billion (21·4-37·4) and $8·0 billion (5·9-10·3), respectively, in India in 2019. This total loss of $36·8 billion (27·4-47·7) was 1·36% of India's gross domestic product (GDP). The economic loss as a proportion of the state GDP varied 3·2 times between the states, ranging from 0·67% (0·47-0·91) to 2·15% (1·60-2·77), and was highest in the low per-capita GDP states of Uttar Pradesh, Bihar, Rajasthan, Madhya Pradesh, and Chhattisgarh. Delhi had the highest per-capita economic loss due to air pollution, followed by Haryana in 2019, with 5·4 times variation across all states. The high burden of death and disease due to air pollution and its associated substantial adverse economic impact from loss of output could impede India's aspiration to be a $5 trillion economy by 2024. Successful reduction of air pollution in India through state-specific strategies would lead to substantial benefits for both the health of the population and the economy. UN Environment Programme; Bill & Melinda Gates Foundation; and Indian Council of Medical Research, Department of Health Research, Ministry of Health and Family Welfare, Government of India.


CRSD: a comprehensive web server for composite regulatory signature discovery.
Transcription factors (TFs) and microRNAs play important roles in the regulation of human gene expression, and the study of their combinatory regulations of gene expression is a new research field. We constructed a comprehensive web server, the composite regulatory signature database (CRSD), that can be applied in investigating complex regulatory behaviors involving gene expression signatures (GESs), microRNA regulatory signatures (MRSs) and TF regulatory signatures (TRSs). Six well-known and large-scale databases, including the human UniGene, mature microRNAs, putative promoter, TRANSFAC, pathway and Gene Ontology (GO) databases, were integrated to provide the comprehensive analysis in CRSD. Two new genome-wide databases, of MRSs and TRSs, were also constructed and further integrated into CRSD. To accomplish the microarray data analysis at one go, several methods, including microarray data pretreatment, statistical and clustering analysis, iterative enrichment analysis and motif discovery, were closely integrated in the web server, which has not been the case in previous studies. Our implementation showed that the published literature could demonstrate the results of genome-wide enrichment analysis. We conclude that CRSD is a powerful and useful bioinformatic web server and may provide new insights into gene regulation networks. CRSD and the online tutorial are publicly available at http://biochip.nchu.edu.tw/crsd1/.


Myorelaxant Activity of essential oil from Origanum majorana L. on rat and rabbit.
Origanum majorana L. (Lamiaceae) was usually used in Moroccan folk medicine to treat infantile colic and abdominal discomfort. The essential oil from the aerial part of the dry Origanum majorana L. (EOOM) was obtained through hydro distillation and analyzed by gas chromatography/mass spectrometry (GC/MS). The effect of EOOM on muscle relaxation was measured on rabbit and rat intestinal smooth muscle mounted in an isotonic transducer. 1) The main compounds obtained from the aqueous extract of this plant were alpha Terpineol, L-terpinen-4-ol and Beta.-Linalool. 2) EOOM inhibited in a concentration-dependent manner spontaneous contraction of rabbit jejunum, with an IC50 = 64.08 ± 2.42 μg/mL. 3) In rat intestine, EOOM induced the relaxation of the tissue in concentration-dependent manner with an IC50 = 39.70 ± 2.29 μg/mL when the tissue was pre-contracted with CCh 10-6 M, and 48.70 ± 2.26 μg/mL when the tissue was pre-contracted with 25 mM KCl. 4) The relaxation effect induced by EOOM was more important than that obtained in the presence of atropine, hexamethonium, Nifedipine, L-NAME and Blue of methylene. the present result indicates that essential oil of Origanum majorana L. exhibit an effect on intestinal relaxation in vitro. This effect further validates the traditional use of Origanum majorana L. to treat infantile colic and abdominal discomfort.


Multilocus statistics to uncover epistasis and heterogeneity in complex diseases: revisiting a set of multiple sclerosis data.
New statistics are developed to gather the contribution of many alleles at different loci to common diseases. Both inferential and descriptive statistics are included in order to uncover epistatic effects as well as heterogeneity. The problem of multiple testing is circumvented by considering a global null hypothesis. Global testing is supplemented by descriptive methods that make use of measures like odds ratio or the P-value of individually tested allele combinations. Visualization helps to reflect complex data sets. The methods described here have been scrutinized by statistical simulations, and we show that power gains can be substantial as compared to single locus statistics. Typing data of multiple sclerosis patients and controls are investigated, representing an example of larger scale information in screening candidate genes for their impact on complex diseases. New insights emerge from this data set demonstrating genetic heterogeneity and evidence for epistasis.


Intrauterine transfusion for parvovirus B19 infection: long-term neurodevelopmental outcome.
To evaluate long-term neurodevelopmental outcome of children treated with intrauterine transfusions for fetal anemia because of parvovirus B19 infection. Children treated with intrauterine transfusions for fetal anemia because of parvovirus B19 infection underwent standardized age-appropriate neurodevelopmental testing. Main outcome was the incidence of neurodevelopmental impairment. Twenty-eight children were evaluated at a median age of 5 years (range, 1.5-13 years). Neurodevelopmental impairment was diagnosed in 3 of 28 (11%) children, including 1 child with combined cerebral palsy and severe developmental delay and 2 children with isolated severe developmental delay. Neurodevelopmental impairment in children treated with intrauterine transfusion for parvovirus B19 infection is increased compared with the general population. Large long-term follow-up studies are required to determine potential risk factors.


Oral anticoagulants as secondary prophylaxis of thrombosis in patients with polycythemia vera: a retrospective analysis of 15 patients.
Thrombotic complications are the leading cause of mortality in patients with polycythemia vera (PV). Phlebotomy and myelosuppressive agents are able to reduce the frequency of thrombosis; however, despite adequate control of myeloproliferation, the thrombotic risk remains increased in PV. This has led to a search for additional antithrombotic agents suited to reduce the rate of thrombotic events in PV. We have retrospectively analyzed 15 patients with PV who received oral anticoagulants (OA) as secondary prophylaxis of thrombosis at our institution with the aim of investigating this treatment approach in patients with PV. Despite OA treatment, 2 (13%) of 15 patients displayed recurrent venous thromboembolism, 3 patients (20%) displayed recurrent arterial thromboembolism, whereas 1 patient (7%) developed recurrent restenosis/occlusion of a transjugular intrahepatic porto-systemic shunt (TIPSS) implanted because of an underlying Budd-Chiari syndrome. Three (20%) of 15 patients had a total of 5 bleeding events during OA treatment, two of which were major bleedings. In none of the major bleedings over-anticoagulation was the reason for bleeding. Within the limitations of a retrospective analysis in a small number of patients, our data suggest that meticulous control of anticoagulant therapy is of critical importance in patients with PV, since in several of our patients recurrent thromboembolic events were associated with a low anticoagulation intensity of INR<2.0. In our opinion, a prospective clinical trial similar to the recently performed ECLAP study would be needed to more thoroughly elucidate the efficacy of OA in the secondary prophylaxis of thrombosis in PV.


Discovery of selective urokinase plasminogen activator (uPA) inhibitors as a potential treatment for multiple sclerosis.
We report here the design and synthesis of a novel series of benzylamines that are potent and selective inhibitors of uPA with promising oral availability in rat. Further evaluation of one representative (ZK824859) of the new structural class showed that this compound lowered clinical scores when dosed in either acute or chronic mouse EAE models, suggesting that uPA inhibitors of this type could be useful for the treatment of multiple sclerosis.


Instability of the tetrathionate respiratory chain of Citrobacter freundii.
The formate--tetrathionate reductase redox pathway of Citrobacter freundii is associated with the cytoplasmic membrane fraction. A high concentration of phosphate in the buffer used for cell disintegration assists in the preparing of membrane particles capable of efficient tetrathionate respiration. A part of this effect at least may be attributed to the high ionic strength of the buffer.


Comprehensive data analysis of human ureter proteome.
Comprehensive human ureter proteome dataset was generated from OFFGel fractionated ureter samples. Our result showed that among 2217 non-redundant ureter proteins, 751 protein candidates (33.8%) were detected in urine as urinary protein/polypeptide or exosomal protein. On the other hand, comparing ureter protein hits (48) that are not shown in corresponding databases to urinary bladder and prostate human protein atlas databases pinpointed 21 proteins that might be unique to ureter tissue. In conclusion, this finding offers future perspectives for possible identification of ureter disease-associated biomarkers such as ureter carcinoma. In addition, Cytoscape GO annotation was examined on the final ureter dataset to better understand proteins molecular function, biological processes, and cellular component. The ureter proteomic dataset published in this article will provide a valuable resource for researchers working in the field of urology and urine biomarker discovery.


Game theory models of functional and aggregative responses.
The expectation of the ideal free distribution is a functional response in which intake is constant in all sites and, if interference is low, an aggregative response in which all individuals cluster in the high prey density sites. Incorporating individual differences in competitive ability results in a functional response in which intake increases with prey density and an aggregative response in which individuals are more evenly distributed.


Measuring propionyl-CoA carboxylase activity in phytohemagglutinin stimulated lymphocytes using high performance liquid chromatography.
Propionyl-CoA carboxylase (PCC) is a mitochondrial enzyme involved in the catabolism of several essential amino acids and odd chain fatty acids. Previous PCC assays have involved either a radiometric assay or have required mitochondria isolation and/or enzyme purification. We developed an enzymatic method to analyze PCC activity in phytohemagglutinin (PHA) stimulated lymphocytes that involves high performance liquid chromatography. The method shows good linearity and sensitivity. PCC activity was unaffected even when lymphocytes were isolated and PHA stimulated after a whole blood sample had been stored at 4°C for 5days. This indicates that this method is suitable for analyzing samples from distant medical centers. The PCC activity of patients with propionic acidemia was found to be much lower than that of normal individuals and carriers. However, this PCC assay is significantly affected by the red blood cell contamination. In conclusion, this is a reliable method for performing PCC assays and only requires 0.5 to 1.0ml of whole blood from newborns. The PCC assay established in this study is useful for the confirmation of PA in individuals, and prenatal diagnosis and genetic counseling for the affected families.


Cancer immunotherapy.
The remarkable specificity of the immune system through antigen recognition has long attracted investigators to the possibility of immune-based therapy for cancer. Previous cancer immunotherapeutics had been restricted to non-specific immunomodulatory agents, such as the cytokines IL-2 or IFN-α. However, the molecular definition of cancer-associated antigens introduced the possibility of specific vaccines and adoptive T cell approaches aiming to target the tumor cells more specifically. The recent introduction of total exome sequencing has enabled the identification of patient tumor-specific epitopes generated through somatic point mutations, raising the possibility of targeting tumor antigens in individual patients which are even more tumor-specific. Transcriptional profiling and immunohistochemistry analyses have revealed a subset of patients with a pre-existing T cell-inflamed tumor microenvironment. This phenotype may be predictive of clinical outcome to immunotherapies and offers the possibility of a predictive biomarker. Further analysis of these tumors has identified a set of defined immune suppressive factors which themselves are being targeted with new immunotherapeutics, already with interesting early phase clinical trial results. Understanding not only the expression of tumor antigens but also the dynamic between a growing tumor and the host immune response is thus generating a rich set of opportunities for the specific immunotherapy of cancer.


Fatty acid composition in fetal, neonatal, and cultured cardiomyocytes in rats.
Reconstructed myocardial tissue still does not have enough pulsatile contraction. It is well known that fetal and mature neonatal cardiomyocytes utilize glucose and lipid, respectively, as their energy substrates, and that cultured ones mainly use glucose in spite of their age comparable to neonate ones, probably due to insufficient supply of lipids from culture medium. In the present study, we compared 7 saturated, 6 monounsaturated, and 11 polyunsaturated fatty acid contents in cultured cardiomyocytes (Cul group) with those in fetal (Fet group, approximately 17 d after impregnation) and neonatal (Neo group, 9 d old) rats, where the age of the Cul cells were set nearly equal to the Neo ones. Saturated fatty acid contents in the Cul group were generally lower than those in the Fet group and were close to those in the Neo group, except for C12:0 of which content was highest in the Neo group. Monounsaturated fatty acid contents in the Cul group were generally lower than those in the Fet group but similar to or higher than those in the Neo group, except for C24:1n-9 of which content was again highest in the Neo group. In contrast, most of polyunsaturated fatty acid (PUFA) contents in the Cul group appeared lower than those in both the Fet and Neo groups, and differences in 5 of 10 detected PUFAs were significant between the Cul and Neo groups. The results suggest that PUFA contents in cultured cardiomyocytes might be insufficient to exert enough contractile ability. In conclusion, it could be necessary for cultured cardiomyocytes to uptake more lipid; PUFAs in particular.


Doublet N-Terminal Oriented Proteomics for N-Terminomics and Proteolytic Processing Identification.
The study of the N-terminome and the precise identification of proteolytic processing events are key in biology. Dedicated methodologies have been developed as the comprehensive characterization of the N-terminome can hardly be achieved by standard proteomics methods. In this context, we have set up a trimethoxyphenyl phosphonium (TMPP) labeling approach that allows the characterization of both N-terminal and internal digestion peptides in a single experiment. This latter point is a major advantage of our strategy as most N-terminomics methods rely on the enrichment of N-terminal peptides and thus exclude internal peptides.We have implemented a double heavy/light TMPP labeling and an automated data validation workflow that make our doublet N-terminal oriented proteomics (dN-TOP) strategy efficient for high-throughput N-terminome analysis.


Effects of photoperiod and temperature on gonadal maturation, and plasma steroid and gonadotropin levels in a cyprinid fish, the honmoroko Gnathopogon caerulescens.
The honmoroko (Gnathopogon caerulescens), a small, spring-summer-breeding cyprinid fish, was exposed to various temperature and photoperiod regimes during different phases of its annual reproductive cycle. In autumn, a clear photoperiodism was seen, especially at warm temperatures (18-24 degrees). The gonadosomatic index, plasma gonadotropin (GtH), and gonadal steroids were higher in fish held at long photoperiod (15L/9D) than in those held at short photoperiod (12L/12D); the differences were greater as the temperature increased. In early spring, gonadal recrudescence progressed at temperatures above 7 degrees, regardless of photoperiod. Photoperiod did not affect plasma GtH and gonadal steroid levels at this time of the year. During its spawning season, a long photoperiod (15L/9D) was again required to maintain gonadal activity and high plasma GtH and gonadal steroid levels, especially at high temperature (25 degrees). In early summer, high temperatures (25 and 30 degrees) suppressed gonadal activity; however, gonadal steroid levels were not depressed, and plasma GtH levels actually increased. The data suggest that under natural conditions, high water temperatures in summer may act directly at the gonadal level, and is probably the factor which terminates the breeding season for this species. The results of these four experiments indicate that the honmoroko exhibits a photoperiodic response during the autumn month and its spawning season, and is dependent on the water temperature. The effects of photoperiod on gonadal maturation appear to be GtH mediated, whereas the suppressive effects of high temperature are not.


MKC-231, a choline uptake enhancer: (2) Effect on synthesis and release of acetylcholine in AF64A-treated rats.
The effect of MKC-231 on acetylcholine (ACh) synthesis and release was studied in the hippocampus of normal and AF64A-treated rats. AF64A (3 nmol/brain, i.c.v.) produced significant reduction of high-affinity choline uptake (HACU) and high K+-induced ACh release in hippocampal synaptosomes. Treatments with MKC-231 (10(-8) and 10(-7) M) showed significant reverse of the decrease in both HACU and ACh release. In hippocampal slices superfused with choline-containing artificial cerebro-spinal fluid (ACSF), high K+-induced ACh release was gradually decreased by repeated alteration of resting and high K+ stimulations in AF64A-treated rats. However, addition of MKC-231 (10(-8) to 10(-7) M) in the superfusate reduces this decrease. In vivo microdialysis studies indicate MKC-231 (10 mg/kg, p.o.) significantly reversed reduction of basal ACh concentrations in AF64A-treated rats, measured by radioimmunoassay without a cholinesterase inhibitor in the perfusate. These results indicate MKC-231 improves AF64A-induced cholinergic hypofunction by enhancing HACU, subsequently facilitating ACh synthesis and release in vitro and in vivo.


The substance P content of peripheral tissues in several mammals.
Levels of substance P immunoreactivity (SPI) were determined in several skin and mucosal areas, in parts of the sympathetic nervous system, the urinary, biliary and respiratory systems of cats, rabbits and guinea-pigs, and in various skin and mucosal areas of humans by radioimmunoassay. Salient findings are (1) The general distribution pattern of SPI in rabbits was similar to that in rodents. (2) The highest SPI tissue levels were found in the sympathetic nervous system, notably in guinea-pigs. (3) The guinea-pig also had the highest SPI levels in ureter, urinary bladder and bile duct. (4) The aorta, pulmonary artery and portal vein of the rabbit contained very low amounts of SPI, the concentration in the carotid sinus being several fold higher. (5) Skin SPI content was generally highest in the cat, especially in the hindpaw-pad, and lowest in abdominal and back skin. (6) SPI levels found in postmortem human skin and mucosal samples are comparable to those found in other mammals. The observations are discussed in view of the sensory innervation of the various tissues.


Identification of a new target muscle for treatment in patients with Parkinson's disease who have lateral trunk flexion?
Parkinson's disease (PD) can present with lateral trunk flexion (LTF). Abnormal posture associated with PD has been treated, but the effectiveness of these treatments is limited, resulting in unsatisfactory outcomes. Unilateral hypertrophy and unilateral hyperactivity may be useful for deciding targets for injection of botulinum toxin or physical rehabilitation. However, such findings may be limited such as the obliquus abdominis muscle or thoracic paraspinal muscles, and several other muscles may have a causative role in LTF. We investigated 8 patients whether other muscles show unilateral hypertrophy by analyzing computed tomographic scans. Cobb's angle was 11° to 34°. The area of the paravertebral muscles was large contralateral to the bending side and this trend intensified from L4 to Th10. The lumbar quadrate muscle and psoas major muscle showed unilateral enlargement. These larger muscles were prominent contralateral to the bending side in five patients and ipsilateral to the bending side in two patients. This unilateral muscle change was mildly seen in the internal and external abdominal oblique muscles. The lumbar quadrate muscle or psoas major muscle showed two hypertrophic patterns, and these muscles might be new therapeutic targets for treatments such as botulinum toxin.


A novel magnetic poly(aniline-naphthylamine)-based nanocomposite for micro solid phase extraction of rhodamine B.
A novel Fe3O4-poly(aniline-naphthylamine)-based nanocomposite was synthesized by chemical oxidative polymerization process as a magnetic sorbent for micro solid phase extraction. The scanning electron microscopy images of the synthesized nanocomposite revealed that the copolymer posses a porous structure with diameters less than 50nm. The extraction efficiency of this sorbent was examined by isolation of rhodamine B, a mutagenic and carcinogenic dye, from aquatic media in dispersion mode. Among different synthesized polymers, Fe3O4/poly(aniline-naphthylamine) nanocomposite showed a prominent efficiency. Parameters including the desorption solvent, amount of sorbent, desorption time, sample pH, ionic strength, extraction time and stirring rate were optimized. Under the optimum condition, a linear spiked calibration curve in the range of 0.35-5.00μgL(-1) with R(2)=0.9991 was obtained. The limits of detection (3Sb) and limits of quantification (10Sb) of the method were 0.10μgL(-1) and 0.35μgL(-1) (n=3), respectively. The relative standard deviation for water sample with 0.5μgL(-1) of RhB was 4.2% (n=5) and the absolute recovery was 92%. The method was applied for the determination of rhodamine B in dishwashing foam, dishwashing liquid, shampoo, pencil, matches tips and eye shadows samples and the relative recovery percentage were in the range of 94-99%.


Location and conformation of amyloid β(25-35) peptide and its sequence-shuffled peptides within membranes: implications for aggregation and toxicity in PC12 cells.
Extracellular deposits of amyloid β (Aβ) aggregates in the brain is the hallmark of Alzheimer's disease. We present the configurations (location and conformation) and the interfacial folding and membrane insertion mechanisms of Aβ fragments, wild-type Aβ(25-35), Aβ(35-25), and a sequence-shuffled peptide [Aβ(25-35)-shuffled] from Aβ(25-35) within membranes by replica-exchange molecular dynamics simulations. Although these peptides have the same amino acid composition, simulations show they have distinct locations and conformations within membranes. Moreover, our in vitro experiments show that these peptides have distinct neurotoxicities. We rationalize the distinct neurotoxicities of these peptides in terms of their simulated locations and conformations in membranes. This work provides another view that complements the general hydrophobicity-toxicity views, to better explain the neurotoxicity of Aβ peptides.


Validation of the Italian version of the Multiple Sclerosis Intimacy and Sexuality Questionnaire-19.
People with multiple sclerosis (MS) may experience sexual dysfunction throughout the disease course. Validated scales to assess sexual dysfunction in MS for Italian patients are lacking. Hence, we aimed at validating Multiple Sclerosis Intimacy and Sexuality Questionnaire (MSISQ-19) for Italian MS patients. We included both male and female MS patients. Each patient completed the Italian translation of the MSISQ-19. Construct validity was explored by the exploratory factor analysis and the Cronbach's alpha coefficient. Test-retest stability and concurrent internal and external validity were examined by Pearson' correlation coefficients. We enrolled 369 MS patients (323 female and 46 male). Italian MSISQ-19 showed a Cronbach's alpha of 0.92. MSISQ-19 test and retest total scores correlated between each other (r = 0.48, p = 0.01). MSISQ-19 total score also correlated with primary, secondary and tertiary subscales (p < 0.001). The Italian Version of the MSISQ-19 showed satisfactory internal consistency and reliability with moderately adequate test-retest reproducibility, suggesting that it may be used as a valuable measure of sexual dysfunction in the Italian population.


Evaluation of extraction approaches linked to ELISA and HPLC for analyses of microcystin-LR, -RR and -YR in freshwater sediments with different organic material contents.
The efficiencies of conventional extraction techniques and analytical methods (HPLC-DAD and ELISA) were investigated for analyses of microcystins (MCs) in sediments. Our results showed several limitations. First, the extraction efficiency strongly depends on the extraction solvent, and extraction with 5% acetic acid in 0.2% trifluoroacetic acid (TFA)-methanol was confirmed as being the most appropriate for three different sediments (recovery: 33.1-44.9% of total MCs according to HPLC analyses). Second, the recovery of MCs was affected by the type of sediment but did not clearly correlate with the content of organic carbon. These results suggest that the sorption of MCs onto inorganic materials such as clay minerals is probably a more important process than interactions of the MCs with organic sediment matter. Third, the structure of the MCs is another crucial factor that affects the sorption of MCs and their recovery from sediments. Hydrophilic MC-RR gave much lower recoveries (20.0-38.8%) than MC-YR (44.1-59.5%) or MC-LR (55.3-77.8%) from three different types of spiked sediments. Recovery results analysed with HPLC-DAD correlated well with ELISA analyses. Further, extraction with 5% acetic acid in 0.2% TFA-methanol was used for analyses of MCs in 34 natural sediment samples collected from Brno reservoir (Czech Republic) from April to October 2005. Concentrations of MCs in sediments ranged from 0.003 to 0.380 microg/g sediment d.m. (ELISA results) or 0.016-0.474 microg/g d.m. (HPLC results). These values are equivalent to 0.63-96.47 microg/L of sediment (ELISA) or 4.67-108.68 microg/L (HPLC), respectively. Concentrations of sediment MCs showed both temporal and spatial variability, with the highest MC contents observed in the spring (April and May) and the lowest concentrations in July and August. Our results demonstrate the suitability of the methods described here for studying the occurrence, fate and ecological role of MCs in the aquatic environment.


Does hormone replacement therapy increase the frequency of breast atypical hyperplasia in postmenopausal women? Results from the Bouches du Rhone district screening campaign.
It is thought that the risk of atypical hyperplasia (AH) increases with age, particularly among postmenopausal women. Three hypotheses were investigated to try to explain this phenomena: use of hormone replacement therapy (HRT), increased breast cancer screening and improvements in radiological quality. Data were collected from the Bouches du Rhône breast cancer screening programme database and from the pathological registry of all women operated on for breast diseases in the district. The AH incidence rate was studied using a Poisson regression analysis. The change in the profile of breast diseases was explored through studying changes in the proportion of AH among benign lesions and malignant diseases. The AH incidence rate significantly increased over time (13.6% per year). The proportion of AH among the benign diseases increased with time and was significantly higher for HRT users (Odds Ratio (OR)=2.05; 95% Confidence Interval (CI): 1.43-2.93). While AH decreased with age among HRT non-users, it increased among users as a proportion of both benign and malignant lesions. The AH incidence rate significantly increased among pre- and postmenopausal women. Our study suggests that this increase is partly explained by the incidental discovery of these lesions by mammography and partly by a real increase of the disease among HRT users.


Negative emotion-related eating behaviours in young women with underweight status, overweight status, anorexia nervosa, and bulimia nervosa in Korea.
This study aimed at investigating the negative emotion-related eating behaviours of young women with extreme weights or eating disorders (EDs). A total of 808 young women participated including 144 with underweight (UW), 364 with normal-weight (NW) and 137 with overweight (OW; including 27 with obesity) from universities, and 63 patients with anorexia nervosa (AN), and 100 patients with bulimia nervosa (BN) from a hospital. Participants were interviewed and completed questionnaires on eating behaviours related to negative emotions. While both OW and BN groups ate more in response to negative emotions, UW and AN groups ate less compared to NW group. Negative feelings after overeating were prominent in OW group and patients with ED. In negative emotions, patients with EDs preferred sweet tastes, but BN group preferred spicy tastes less. Patients with EDs had increased external eating, and BN group also showed increased restrained eating. Neuroticism had indirect impacts on OW status via emotional eating unlike BN. The results suggest that UW and OW status were on the spectrum of emotional eating, and AN and BN were on the extremes of external eating, which may be integrated into the development of interventions targeting each status.


Fiber-optic infrared reflection absorption spectroscopy for trace analysis on surfaces of varying roughness. Part II: Acetaminophen on stainless steel.
Investigations of the effects of surface roughness on the utility of grazing-angle Fourier transform infrared reflection absorption spectroscopy (IRRAS) as a method for quantifying trace contamination of metal surfaces have been extended to acetaminophen, a model active pharmaceutical agent, on 316 stainless steel. The effects are more complicated than for the surfactant sodium dodecyl sulfate (SDS) on stainless steel; they include a strong surface-finish dependence of sensitivity and nonlinear behavior at surface loadings above approximately 1-2 microg cm(-2). Using data from samples in the loading range 0-0.5 microg cm(-2), unbiased partial least squared calibrations can be readily achieved for individual surface finishes with detection limits of L(D) approximately 0.15 microg cm(-2). However, as found for SDS on stainless steel, models built using data from samples of mixed surface roughness are more problematic.


Thiazolidione derivatives targeting the histidine kinase YycG are effective against both planktonic and biofilm-associated Staphylococcus epidermidis.
To evaluate the efficacies of six derivatives of Compound 2, a novel YycG histidine kinase inhibitor with the thiazolidione core structure in the treatment of medical device-related biofilm infections. The minimal inhibitory concentration (MIC) of the derivatives was determined using the macrodilution broth method, and the minimal bactericidal concentration (MBC) was obtained via sub-culturing 100 μL from each negative tube from the MIC assay onto drug-free Mueller-Hinton agar plates. Biofilm-killing effect for immature (6 h-old) biofilms was examined using a semiquantitative plate assay, and the effect on mature (24 h-old) biofilms was observed under a confocal laser scanning microscope (CLSM). The derivatives potently suppressed the growth of Staphylococcus epidermidis. The MIC values of the derivatives H2-10, H2-12, H2-20, H2-29, H2-27, and H2-28 on S epidermidis ATCC 35984 were 24.3, 6.5, 6.2, 3.3, 3.1, and 1.5 μg/mL, respectively. The MBC values of these derivatives were 48.6, 52.2, 12.4, 52.6, 12.4, and 6.2 μg/mL, respectively. The derivatives killed all bacteria in immature (6 h-old) biofilms and eliminated the biofilm proliferation. The derivatives also displayed strong bactericidal activities toward cells in mature (24 h-old) biofilms, whereas they showed low cytotoxicity and hemolytic activity toward Vero cells and human erythrocytes. The bactericidal and biofilm-killing activities of the new anti-YycG compounds were significantly better than the parent Compound 2.


Electrochemical Oxidative Esterification of Thiophenols: Efficient Access to Sulfinic Esters.
The sulfinic esters are important and useful building blocks in organic synthesis. The aim of this study was to develop a simple and efficient method for the synthesis of sulfinic esters. Constant current electrolysis from thiols and alcohols was selected as the method for the synthesis of sulfinic esters. A novel electrochemical method for the synthesis of sulfinic esters from thiophenols and alcohols has been developed. Up to 27 examples of sulfinic esters have been synthesized using the current methods. This protocol shows good functional group tolerance as well as high efficiency. In addition, this protocol can be easily scaled up with good efficiency. Notably, heterocycle-containing substrates, including pyridine, thiophene, and benzothiazole, gave the desired products in good yields. A plausible reaction mechanism is proposed. This research not only provides a green and efficient method for the synthesis of sulfinic esters but also shows new applications of electrochemistry in organic synthesis. It is considered that this green and efficient synthetic protocol used to prepare sulfinic esters will have good applications in the future.


Use of the external fixation apparatus for percutaneous insertion of pins in the distal one-third of the radius: an anatomic study.
To assess the risk of soft-tissue injury during percutaneous placement of external fixation pins in the proximal radius. An anatomic study with embalmed cadaver limbs. Hand and upper limb centre at a university-affiliated hospital. Two 4-mm Hoffman half pins were percutaneously placed along the dorsoradial ridge of the radius, four finger breadths proximal to the radial styloid process. Injuries to soft tissues including tendons, nerves and vessels were noted. Nerve or tendon injuries occurred in 7 of 26 forearms. Three pins transfixed either the superficial branch of the radial nerve or lateral antebrachial cutaneous nerves. Tendon injuries included the brachioradialis in two forearms, the extensor carpi radialis brevis in three forearms, and the extensor carpi radialis longus and the abductor pollicis longus in one forearm each. Percutaneous pin placement in the distal radius is unsafe. The authors recommend open pin placement for fractures of the distal radius.


Hereditary multiple exostoses and enchondromatosis.
Hereditary multiple exostoses (HME) is an autosomal-dominant disorder characterized by the development of benign tumours, multiple osteochondromas (exostoses), growing outward from the metaphyses of long bones. Birth prevalence is estimated to be one in 50,000, and the severity of the disease is variable. Osteochondromas may cause complications including pain, deformities and shortening of the long bones, restricted motion of joints, nerve or blood vessel compression, and malignant transformation (5% of cases) in adulthood. HME is a genetically heterogeneous disorder and is associated with mutations in EXT1 or EXT2 genes, which are both tumour suppressor genes. EXT genes encode glycosyltransferases, termed 'exostosins', which are involved in the biosynthesis of heparan sulphate. Enchondromatosis (or Ollier disease) is characterized by the presence of intra-osseous benign cartilaginous tumours. The estimated prevalence of the disease is one in 100,000. An asymmetrical distribution of cartilage lesions is observed in the disease. The number, size and location of the enchondromas can be extremely variable between patients. Clinical problems caused by enchondromas include skeletal deformities, limb length discrepancy, pain and the potential risk for malignant change to chondrosarcoma (20-50% of cases). The condition in which multiple enchondromas is associated with haemangiomas is known as 'Maffucci syndrome'. Ollier disease and Maffucci syndrome are not usually inherited disorders.


Chronic nerve compression alters Schwann cell myelin architecture in a murine model.
Myelinating Schwann cells compartmentalize their outermost layer to form actin-rich channels known as Cajal bands. Herein we investigate changes in Schwann cell architecture and cytoplasmic morphology in a novel mouse model of carpal tunnel syndrome. Chronic nerve compression (CNC) injury was created in wild-type and slow-Wallerian degeneration (Wld(S) ) mice. Over 12 weeks, nerves were electrodiagnostically assessed, and Schwann cell morphology was thoroughly evaluated. A decline in nerve conduction velocity and increase in g-ratio is observed without early axonal damage. Schwann cells display shortened internodal lengths and severely disrupted Cajal bands. Quite surprisingly, the latter is reconstituted without improvements to nerve conduction velocity. Chronic entrapment injuries like carpal tunnel syndrome are primarily mediated by the Schwann cell response, where decreases in internodal length and myelin thickness disrupt the efficiency of impulse propagation. Restitution of Cajal bands is not sufficient for remyelination after CNC injury.


Percutaneous transhepatic cholangiography in diagnostic evaluation of 160 jaundiced patients. Results of an improved technic.
Percutaneous transhepatic cholangiography (PTC) was carried out in 160 patients with jaundice in whom the diagnosis could not be established by means of conventional methods of investigation. In PTC, selective catheterization of the common bile duct was employed and cholangiograms of high quality were obtained. In patients with obstruction of the biliary passages, the catheter was left indwelling centrally in the biliary passages for external bile drainage. The investigation, thus, did not necessitate immediate operation in these patients. PTC was successful in 113 of the 115 patients with obstructive jaundice. In forty-five patients the suspicion of obstruction could be discharged. Normal biliary passages were demonstrated in twenty-six patients. In nineteen patients it was impossible to cannulate the biliary tree and this was tantamount to nonobstructive jaundice. Complications occurred in six patients, but were not exclusively due to PTC. PTC provides a possibility for differentiation between obstructive and nonobstructive jaundice and thereby laparotomy can frequently be avoided. The incidence of complications may be maintained at an acceptable, low level provided the technic is meticulously observed.


Real-World Biomarker Test Utilization and Subsequent Treatment in Patients with Early-Stage Non-small Cell Lung Cancer in the United States, 2011-2021.
Biomarker testing is increasingly crucial for patients with early-stage non-small cell lung cancer (eNSCLC). We explored biomarker test utilization and subsequent treatment in eNSCLC patients in the real-world setting. Using COTA's oncology database, this retrospective observational study included adult patients ≥ 18 years old diagnosed with eNSCLC (disease stage 0-IIIA) between January 1, 2011 and December 31, 2021. Date of first eNSCLC diagnosis was the study index date. We reported testing rates by index year for patients who received any biomarker test within 6 months of eNSCLC diagnosis and by each molecular marker. We also evaluated treatments received among patients receiving the five most common biomarker tests. Among the 1031 eNSCLC patients included in the analysis, 764 (74.1%) received ≥ 1 biomarker test within 6 months of eNSCLC diagnosis. Overall, epidermal growth factor receptor (EGFR; 64%), anaplastic lymphoma kinase (ALK; 60%), programmed death receptor ligand 1 (PD-L1; 48%), ROS proto-oncogene 1 (ROS1; 46%), B-Raf proto-oncogene (40%), mesenchymal epithelial transition factor receptor (35%), Kirsten rat sarcoma viral oncogene (29%), RET proto-oncogene (22%), human epidermal growth factor receptor 2 (21%), and phosphatidylinositol-4,5-bisphosphate 3-kinase catalytic subunit alpha (20%) were the 10 most frequently tested biomarkers. The proportion of patients undergoing biomarker testing rose from 55.3% in 2011 to 88.1% in 2021. The most common testing methods were Sanger sequencing for EGFR (244, 37%), FISH (fluorescence in situ hybridization) for ALK (464, 75%) and ROS1 (357, 76%), immunohistochemical assay for PD-L1 (450, 90%), and next-generation sequencing testing for other biomarkers. Almost all the 763 patients who received the five most common biomarker tests had a test before the initiation of a systemic treatment. This study suggests a high biomarker testing rate among patients with eNSCLC in the US, with testing rates for various biomarkers increasing over the past decade, indicating a continuous trend towards the personalization of treatment decisions.


Enhancement of viral and DNA mediated transformation of cloned rat embryo fibroblast cells by 3-aminobenzamide.
We have analyzed the effect of the poly(ADP-ribose) synthesis inhibitor 3 aminobenzamide (3AB) on de novo and methyl methanesulfonate (MMS) and gamma irradiation enhancement of viral transformation of a cloned rat embryo fibroblast cell line, CREF, by a cold-sensitive host-range mutant of type 5 adenovirus, H5hr1. Additionally, we have evaluated the effect of 3AB on the transformation of CREF cells following transfection with a gene conferring resistance to hygromycin (hygr) or the neomycin analogue G418 (neor) in combination with a cloned type 5 adenovirus E1A transforming gene (Ad5 E1A) or the Ha-ras (T24) oncogene. 3AB induced a dose- and time-dependent increase in the level of de novo MMS-enhanced and gamma irradiation-enhanced transformation of CREF cells by H5hr1, whereas it did not induce morphological transformation in uninfected control, MMS-pretreated, or gamma irradiation-pretreated CREF cells. Temporal kinetic studies indicated that 3AB was most effective in enhancing de novo and MMS-enhanced and gamma irradiation-enhanced viral transformation when applied early after viral and carcinogen plus viral infection and when present for extended time periods (4-5 wk). 3AB also increased the frequency of resistant colonies following transfection with several cloned genes, including hygr, neor, and protein kinase C (which also contained a neor gene), and the frequency of morphological transformation of CREF cells following cotransfection with a hygr gene and an Ad5 E1A or an activated Ha-ras (T24) gene. In contrast, 3AB exerted the opposite effect, i.e., an inhibitory effect, when applied to NIH 3T3 cells transfected with a hygr or neor gene, alone or in combination with a T24 gene. The ability of 3AB to enhance the frequency of transformation of CREF cells was not associated with a selective effect on the growth of H5hr1-transformed CREF cells in monolayer or agar culture. Similarly, 3AB did not alter the percentage of MMS- or gamma irradiated-pretreated H5hr1-infected cells retaining free Ad5 DNA or the random pattern or quantity of viral DNA integration in control or carcinogen-treated H5hr1-transformed cells. These results suggest that cellular processes regulated by the nuclear enzyme ADPRT, or additional processes modified by 3AB, may be important mediators of stable transformation induced by transfected DNA and both de novo and carcinogen-enhanced viral transformation of specific target cells.


In Vitro Investigation of Influences of Chitosan Nanoparticles on Fluorescein Permeation into Alveolar Macrophages.
Pulmonary infection namely tuberculosis is characterized by alveolar macrophages harboring a large microbe population. The chitosan nanoparticles exhibit fast extracellular drug release in aqueous biological milieu. This study investigated the matrix effects of chitosan nanoparticles on extracellular drug diffusion into macrophages. Oligo, low, medium and high molecular weight chitosan nanoparticles were prepared by nanospray drying technique. These nanoparticles were incubated with alveolar macrophages in vitro and had model drug sodium fluorescein added into the same cell culture. The diffusion characteristics of sodium fluorescein and nanoparticle behavior were investigated using fluorescence microscopy, scanning electron microscopy, differential scanning calorimetry and Fourier transform infrared spectroscopy techniques. The oligochitosan nanoparticles enabled macrophage membrane fluidization with the extent of sodium fluorescein entry into macrophages being directly governed by the nanoparticle loading. Using nanoparticles made of higher molecular weight chitosan, sodium fluorescein permeation into macrophages was delayed due to viscous chitosan diffusion barrier at membrane boundary. Macrophage-chitosan nanoparticle interaction at membrane interface dictates drug migration into cellular domains.


A direct spectrophotometric assay for D-alanine carboxypeptidases and for the esterase activity of beta-lactamases.
A direct spectrophotometric pH indicator method has been devised to assay the activity of the D-Ala carboxypeptidase/transpeptidase of Streptomyces R61, and which should be of general application to D-Ala carboxypeptidases. The substrate employed is N,N'-diacetyl-L-lysyl-D-alanyl-D-lactate. The method allows the determination of steady-state kinetic parameters, and can also be used for the assay of the esterase activity of beta-lactamases against specific depsipeptides.


Geospatial tools and Chagas control: a critical comment on the paper of Weinberg et al., Evaluation and planning of Chagas control activities using geospatial tools. Geospat Health 2019;14:229-238.
The contribution of the geospatial tools to the vector control activities of Chagas disease is very clear for the academic and technology-prone communities, but not so much for the state-dependent health agents (national or provincial) that are ultimately responsible for disease control in Latin America. (...).


Synthesis of novel polyphenols consisted of ferulic and gallic acids, and their inhibitory effects on phorbol ester-induced Epstein-Barr virus activation and superoxide generation.
We prepared novel polyphenols which were esters composed of two naturally occurring products, ferulic and gallic acids, and investigated their inhibitory effects on 12-O-tetradecanoylphorbol-13-acetate (TPA)-induced Epstein-Barr virus (EBV) activation and superoxide (O2-) generation. Most of these compounds exhibited significant EBV activation suppression at a concentration of 20 microM and in particular, the ester 5f having 2-methyl-1-butyl group showed high activity. The suppressive effects on O2- generation were also observed in most of the esters.


Cutaneous mastocytosis in children: anaesthetic considerations.
Despite considerable information and discussion about the risk of serious complications in patients with systemic mastocytosis undergoing general anaesthesia, little is known specifically about the risk to patients with isolated cutaneous mastocytosis. The experience of 29 general anaesthetics in 12 children with urticaria pigmentosa and three with solitary cutaneous mastocytoma was reviewed. No major complications were encountered and the four minor problems seen were self-limiting. The data from this study do not suggest that patients with urticaria pigmentosa or solitary cutaneous mastocytoma are at increased risk of life-threatening complications under general anaesthesia.


Tympanic membrane retraction: An endoscopic evaluation of staging systems.
The objectives of this work were to assess inter- and intraobserver variability of different staging systems for tympanic membrane (TM) retraction using otoendoscopy in children at risk of retraction from cleft palate, to compare hearing level with stage of retraction, and to propose optimum characteristics for monitoring TM retraction with endoscopy. Cross-sectional study. Endoscopic images of 245 TMs of children with cleft palate (mean age, 13.0 years) were assessed on two separate occasions by six observers using the Sade and Erasmus staging systems for pars tensa retraction and Tos system for pars flaccida retraction. Intra- and interobserver agreements were calculated. Extent of TM retraction was compared with hearing threshold. TMs with middle ear effusion, tympanostomy tubes, or perforation were excluded. A total of 108 ear drums (44%) were rated as having pars tensa and/or flaccida retraction. Intraobserver agreement was fair to moderate (kappa = 0.3-0.37, P < .001) for the different staging systems and interobserver agreement slight to moderate (0.18-0.41 P < .001). Conductive hearing loss (four-tone average air-bone gap >25 dB HL) was present in 11 ears (15%). No correlation between hearing threshold and retraction stage was found. Isolated tensa retraction onto the promontory increased hearing threshold more than retraction involving the incus (P = .02; analysis of variance). Endoscopic image capture may provide a clear objective record of TM retraction, but current staging systems have unsatisfactory reliability when applied to such images, and retraction stage correlates poorly with hearing threshold. Modification of retraction assessment to improve validity and clinical relevance is proposed.


One-Year Follow-Up of Healthy Older Adults with Electroencephalographic Risk for Neurocognitive Disorder After Neurofeedback Training.
In healthy older adults, excess theta activity is an electroencephalographic (EEG) predictor of cognitive impairment. In a previous study, neurofeedback (NFB) treatment reinforcing reductions theta activity resulted in EEG reorganization and cognitive improvement. To explore the clinical applicability of this NFB treatment, the present study performed a 1-year follow-up to determine its lasting effects. Twenty seniors with excessive theta activity in their EEG were randomly assigned to the experimental or control group. The experimental group received an auditory reward when the theta absolute power (AP) was reduced. The control group received the reward randomly. Both groups showed a significant decrease in theta activity at the training electrode. However, the EEG results showed that only the experimental group underwent global changes after treatment. These changes consisted of delta and theta decreases and beta increases. Although no changes were found in any group during the period between the posttreatment evaluation and follow-up, more pronounced theta decreases and beta increases were observed in the experimental group when the follow-up and pretreatment measures were compared. Executive functions showed a tendency to improve two months after treatment which became significant one year later. These results suggest that the EEG and behavioral benefits of this NFB treatment persist for at least one year, which adds up to the available evidence contributing to identifying factors that increase its efficacy level. The relevance of this study lies in its prophylactic features of addressing a clinically healthy population with EEG risk of cognitive decline.


Analysis of brain signal change response in amygdala evoked by skin pressure stimulus.
It was well known that the human body would produce an uncomfortable sensation when the fabric exerted a certain amount of pressure irritation on the skin. The amygdala had long been thought to be the source of negative emotion perception. However, up to now, the brain signal changes in the amygdala evoked by skin exposure pressure had not been known. In this work, a series of gradually increasing contact pressure stimulus from boneless corsets was repeatedly applied to the body's waist and abdomen, and the technology of functional magnetic resonance imaging (fMRI) was adopted to detect the brain response synchronously. The results shown that both subjective comfort score and percent signal changes (PSCs) of amygdala decreased with the increase of skin contact pressure. When the skin pressure applied to the waist and abdomen of the human body exceeded about 1 kPa, blood oxygen level dependent signal in the amygdala was negatively activated. Besides, the degree of response of PSCs was intense than subjective evaluation, and the standard deviations of PSCs between individuals were much smaller than subjective evaluations. It was suggested that skin contact pressure stimulus caused the attention of the amygdala brain area. The greater the stimulus, the higher the attention, but such attention was caused by negative activation of the amygdala induced by skin discomfort. In addition, skin comfort representation based on brain perception was superior to subjective representation due to its higher response sensitivity and antipsychological interference ability.


Embryonal Origin of MTSCC of Kidney May Explain its Morphological Heterogeneity: Diagnostic Impact of Genetic Analysis.
Previous genetic and morphologic characterisation of mucinous tubular and spindle cell carcinoma (MTSCC) have yielded controversial results. The aim of this study was to explain the phenotypic heterogeneity of MTSCC diagnosed by genetic means. We analyzed 7 MTSCC by array CGH and microsatellite allelotyping and by histology for morphological variation. We worked-up two entire kidneys with MTSCC to find microscopic alterations. We confirmed the diagnosis of MTSCC by detecting copy number changes at chromosomes 1, 4, 6, 8, 13, 14, 15, 18 and 22. We detected 13 small, microscopic precursor lesions in the two kidneys and found similar histological structures in precursor lesions and MTSCC. MTSCC develops from embryonal rest-like lesions of impaired differentiation which may explain its morphological variations. Until diagnosis of a "malignant" MTSCC" is not confirmed by genetic means, it should not be called carcinoma.


Calcium-myristoyl Tug is a new mechanism for intramolecular tuning of calcium sensitivity and target enzyme interaction for guanylyl cyclase-activating protein 1: dynamic connection between N-fatty acyl group and EF-hand controls calcium sensitivity.
Guanylyl cyclase-activating protein 1 (GCAP1), a myristoylated Ca(2+) sensor in vision, regulates retinal guanylyl cyclase (RetGC). We show that protein-myristoyl group interactions control Ca(2+) sensitivity, apparent affinity for RetGC, and maximal level of cyclase activation. Mutating residues near the myristoyl moiety affected the affinity of Ca(2+) binding to EF-hand 4. Inserting Phe residues in the cavity around the myristoyl group increased both the affinity of GCAP1 for RetGC and maximal activation of the cyclase. NMR spectra show that the myristoyl group in the L80F/L176F/V180F mutant remained sequestered inside GCAP1 in both Ca(2+)-bound and Mg(2+)-bound states. This mutant displayed much higher affinity for the cyclase but reduced Ca(2+) sensitivity of the cyclase regulation. The L176F substitution improved affinity of myristoylated and non-acylated GCAP1 for the cyclase but simultaneously reduced the affinity of Ca(2+) binding to EF-hand 4 and Ca(2+) sensitivity of the cyclase regulation by acylated GCAP1. The replacement of amino acids near both ends of the myristoyl moiety (Leu(80) and Val(180)) minimally affected regulatory properties of GCAP1. N-Lauryl- and N-myristoyl-GCAP1 activated RetGC in a similar fashion. Thus, protein interactions with the central region of the fatty acyl chain optimize GCAP1 binding to RetGC and maximize activation of the cyclase. We propose a dynamic connection (or "tug") between the fatty acyl group and EF-hand 4 via the C-terminal helix that attenuates the efficiency of RetGC activation in exchange for optimal Ca(2+) sensitivity.


Use of eculizumab in children with allogeneic haematopoietic stem cell transplantation associated thrombotic microangiopathy - a multicentre retrospective PDWP and IEWP EBMT study.
Terminal complement blockade by humanised monoclonal antibody eculizumab has been used to treat transplantation-associated thrombotic microangiopathy (TA-TMA) in recent years. This retrospective international study conducted by the Paediatric Diseases (PDWP) and Inborn Error Working Party (IEWP) of the European Society for Blood and Marrow Transplantation (EBMT) describes outcome and response of 82 paediatric patients from 29 centres who developed TA-TMA and were treated with eculizumab between January 2014 and May 2019. The median time from hematopoietic stem cell transplantation (HSCT) to TA-TMA manifestation was 92 days (range: 7-606) and from TA-TMA diagnosis to the start of eculizumab treatment 6 days (range: 0-135). Most patients received eculizumab weekly (72%, n = 55) with a standard weight (kg)-based dose (78%, n = 64). Six months from beginning of eculizumab therapy, the cumulative incidence of TA-TMA resolution was 36.6% (95% CI: 26.2-47) and the overall survival (OS) was 47.1% (95% CI: 35.9-57.5). All 43 patients with unresolved TA-TMA died. The cause of death was HSCT-related in 41 patients. This study also documents poor outcome of patients without aGvHD and their frequent concomitant viral infections. Considering recent publications, intensified eculizumab dosing and complement monitoring could potentially improve upon outcomes observed in this study.


Preparation, characterization, and in vitro evaluation of physostigmine-loaded poly(ortho ester) and poly(ortho ester)/poly(D,L-lactide-co-glycolide) blend microspheres fabricated by spray drying.
The physostigmine-loaded poly(ortho ester) (POE), poly(dl-lactide-co-glycolide) (PLGA) and POE/PLGA blend microspheres were fabricated by a spray drying technique. The in vitro degradation of, and physostigmine release from, the microspheres were investigated. SEM analysis showed that the POE and POE/PLGA blend particles were spherical. They were better dispersed when compared to the pure PLGA microspheres. Two glass transition temperature ( Tg ) values of the POE/PLGA blend microspheres were observed due to the phase separation of POE and PLGA in the blend system. XPS analysis proved that POE dominated the surfaces of POE/PLGA blend microspheres, indicating that the blend microspheres were coated with POE. The encapsulation efficiencies of all the microspheres were more than 95%. The incorporation of physostigmine reduced the Tg value of microspheres. The Tg value of the degrading microspheres increased with the release of physostigmine. For instance, POE blank microspheres and physostigmine-loaded POE microspheres had a Tg value of 67 degrees C and 48 degrees C, respectively. After 19 days in vitro incubation, Tg of the degrading POE microspheres increased to 55 degrees C. Weight loss studies showed that the degradation of the blend microspheres was accelerated with the presence of PLGA because its degradation products catalyzed the degradation of both POE and PLGA. The release rate of physostigmine increased with increase of PLGA content in the blend microspheres. The initial burst release of physostigmine was effectively suppressed by introducing POE to the blend microspheres. However, there was an optimized weight ratio of POE to PLGA (85:15 in weight), below which a high initial burst was induced. The POE/PLGA blend microspheres may make a good drug delivery system.


Treatment with imatinib was useful to delay the neointimal hyperplasia of aortocaval fistula in adenine-induced renal failure rats.
The study was conducted to investigate the effect of the treatment with imatinib, a c-kit specific inhibitor, on the neointimal hyperplasia (NIH) of aortocaval fistula (ACF) in adenine-induced renal failure rats. All rats were randomly assigned to 4 groups: rats were fed on a normal diet (normal group); rats were fed on a 0.75% adenine-rich diet (renal failure group). The remaining rats underwent ACF after receiving a 0.75% adenine-rich diet and received daily saline gavage (model group) or imatinib gavage (imatinib group) for 7 days after surgery. Immunohistochemical method was used to detect c-kit expression, and Elastomeric Verhoeff-Van Gieson (EVG) staining was used to observe morphological changes of the ACF. The Pearson correlation analysis was used to evaluate the correlations of c-kit expression with intimal thickness and the percentage of stenosis, respectively. The renal failure group showed positive c-kit expression on the intima of the inferior vena cava (IVC), whereas the normal group did not. Compared to the model group, intimal thickness (P = 0.001), the percentage of stenosis (P = 0.006) and c-kit expression (P = 0.04) were decreased in the imatinib group at 8 weeks postoperatively. C-kit expression was positively correlated with both intimal thickness and percentage of stenosis (intimal thickness: R = 0.650, P = 0.003; the percentage of stenosis: R = 0.581, P = 0.011) in both the model and imatinib groups. Treatment with imatinib, a c-kit specific inhibitor, was useful to delay the NIH of ACF in adenine-induced renal failure rats.


Revision of a Fractured Titanium Modular Revision Hip Stem Without Removal of the Well-Fixed Part of the Stem: A New Approach Using a Cemented Tube.
Component fracture is a rare cause for revision in total hip arthroplasty. For a fractured well-fixed long femoral stem, the options are limited. We sought to develop a technique to address this problem with lower morbidity. A newly developed cemented tube was constructed and cemented onto a fractured Revitan revision hip femoral stem to retain the distal well-fixed component. At the 2-year follow-up, the Harris Hip Score, pain level, and radiographic images were analyzed. At the 2-year follow-up, no radiological signs of loosening or failure could be observed. The patient's preoperative Harris Hip Score improved from 42.8 to 97 points. The pain level improved from 7/10 to 0/10. Our case report depicts excellent clinical and radiographic outcomes at 2-year follow-up by a newly developed cemented tube technique. This technique is a potential new option for revision of fractured well-fixed diaphyseal stems without major bone loss. Our successful results suggest this technique is worthy of consideration and further study.


The effects of adenosine on the development of long-term potentiation.
In previous work we found that a brief period of hypoxia occurring within 1-2 min of high-frequency stimulation disrupts the development of long-term potentiation (LTP) in the CA1 field of rat hippocampal slices. We now report that extracellular application of adenosine if applied within 1 min but not 5 min after high-frequency stimulation similarly prevents LTP. Adenosine was ineffective if DPCPX (1,3-dipropyl-8-cyclopentyl-xanthine), a selective antagonist of adenosine A1 receptors, was present. The post-stimulation application mode excludes the possibility that adenosine interferes with NMDA receptor activation and its role in initiating LTP. It suggests instead that changes in intracellular control systems linked to adenosine receptors can during a brief vulnerable period interrupt the biochemical processes leading to the expression of long-term potentiation.


Functional mapping imprinted quantitative trait loci underlying developmental characteristics.
Genomic imprinting, a phenomenon referring to nonequivalent expression of alleles depending on their parental origins, has been widely observed in nature. It has been shown recently that the epigenetic modification of an imprinted gene can be detected through a genetic mapping approach. Such an approach is developed based on traditional quantitative trait loci (QTL) mapping focusing on single trait analysis. Recent studies have shown that most imprinted genes in mammals play an important role in controlling embryonic growth and post-natal development. For a developmental character such as growth, current approach is less efficient in dissecting the dynamic genetic effect of imprinted genes during individual ontology. Functional mapping has been emerging as a powerful framework for mapping quantitative trait loci underlying complex traits showing developmental characteristics. To understand the genetic architecture of dynamic imprinted traits, we propose a mapping strategy by integrating the functional mapping approach with genomic imprinting. We demonstrate the approach through mapping imprinted QTL controlling growth trajectories in an inbred F2 population. The statistical behavior of the approach is shown through simulation studies, in which the parameters can be estimated with reasonable precision under different simulation scenarios. The utility of the approach is illustrated through real data analysis in an F2 family derived from LG/J and SM/J mouse stains. Three maternally imprinted QTLs are identified as regulating the growth trajectory of mouse body weight. The functional iQTL mapping approach developed here provides a quantitative and testable framework for assessing the interplay between imprinted genes and a developmental process, and will have important implications for elucidating the genetic architecture of imprinted traits.


Antigen-specific antibody production of human B cells in NOG mice reconstituted with the human immune system.
Passive antibody administration shows strong potential as a new therapeutic method. In clinical applications, human-derived antibodies with antigen specificity are more useful without putting individuals at risk. Production of human-derived antibodies against given antigens can be obtained from animal models if the human immune system is established in the animals. In fact, past reports revealed that human T and B cells develop from hematopoietic progenitor cells in immunodeficient mice. However, there have been few reports on sufficient induction of antigen-specific antibodies, particularly IgG, in immunodeficient mice reconstituted with human immune cells. In this chapter, we discuss a major shortcoming of induction of antigen-specific IgG antibodies in human immune cells developed in the murine environment based on our data. We demonstrated that human T cell development is restricted by the murine MHC and consequently T cells may not achieve cognate interaction with human B cells. Human B cells developed in the mouse are mainly CD5+B1 cells that preferentially produce IgM. At the same time, human LN transplantation on the spleen enabled NOG mice to produce antigen-specific IgG antibody. These results suggest that if efficient cognate interaction mediated by a certain antigen on MHC class II between human T and B-2 cells occurs, human B cells can produce IgG antibody against a given antigen in the murine environment.


A Comparison of the Open Semi-extended Parapatellar Versus Standard Entry Tibial Nailing Techniques and Knee Pain: A Randomized Controlled Trial.
To determine if an open, semi-extended, parapatellar tibial nailing technique (SEK) imparts any undue knee symptoms/pain compared with the traditional infrapatellar tibial nailing technique (FK). Randomized controlled trial. Level 1 trauma center. Forty-seven patients with OTA/AO 42A-C tibial shaft fractures were included in the study. Patients were randomized to treatment with either a SEK or an FK technique. The main outcome was a symptoms subset of the International Knee Documentation Committee (SS-IKDC) pertaining to knee symptoms/pain. Final follow-up was collected at 1 year for 24 SEK and 23 FK patients. No significant differences were found between the groups in regards to demographics, injury, or surgery-related variables. The 2 techniques did have equivalent symptoms scores [mean for the difference (SEK - FK): 0.29, 90% confidence interval: -4.16 to 4.75] but did not have equivalent pain scores [mean for the difference (SEK - FK): 0.2, 90% confidence interval: -3.18 to 3.59]. When comparing demographic and injury-related variables to the SS-IKDC, only Kellgren-Lawrence classification was statistically significant (P = 0.026), where increasing presence of osteoarthritis was associated with lower scores. The results of this single-center randomized controlled trial show that SEK and FK techniques for tibial nailing are equivalent in regards to knee symptoms (defined as ±5 points on the SS-IKDC) but not specifically pain that showed trends toward decreasing knee pain with the open SEK technique. This randomized controlled trial demonstrates that the use of the open semi-extended, parapatellar technique for tibial nailing should not be associated with any higher likelihood of knee pain/symptoms than the traditional flexed knee, infrapatellar technique. Therapeutic Level I. See Instructions for Authors for a complete description of levels of evidence.


Effect of a traditional Japanese herbal medicine, hochu-ekki-to (Bu-Zhong-Yi-Qi Tang), on immunity in elderly persons.
In general, the elderly show a significant age-related decline in their immune response, thus leading to an increased vulnerability to infections or to an increase in the occurrence of malignant tumors. In this study, we examined the effect of Hochu-ekki-to (HOT or TJ-41) on the immunological capacity of the elderly. A group of elderly patients complaining of general fatigue or weakness were orally administered 7.5 g of HOT everyday for at least 120 days (4 months), whereas another group of aged patients mainly complaining of a loss of appetite were daily given 7.5 g of Anchu-san (TJ-5) during the same period and served as a control group. From the immunological point of view, the total number of circulating leukocytes remained unchanged, during the observation period both in the HOT and Anchu-san groups, as well as the ratios between CD3(+) T and CD20(+) B cells and between CD4(+) T and CD8(+) T cells. In addition, no differences were observed in the expression of CD25 antigen, which represents an activated state of T cells. However, as verified on day 30 as well as on day 120 after the administration of HOT, the natural killer (NK) activity against K562 target cells was significantly enhanced, in comparison to the results on day 0 in the HOT group, as well as to that activity on days 0, 30 and 120 in the Anchu-san group. In addition, on days 30 and 120 in the HOT group, there was a significant increase in the serum IFN-gamma level, which is thought to be associated with the NK activity, whereas no significant changes in that level were observed in the Anchu-san group, during the study period. From these results, it may be concluded that the administration of HOT to elderly people may help them improve, at least to some degree, their immunological capacity.