Kaposi's sarcoma-associated herpesvirus K8 exon 3 contains three 5'-splice sites and harbors a K8.1 transcription start site.
Kaposi's sarcoma-associated herpesvirus (KSHV) K8 and K8.1 open reading frames are juxtaposed and span from nucleotide (nt) 74850 to 76695 of the virus genome. A K8 pre-mRNA overlaps the entire K8.1 coding region, and alternative splicing of KSHV K8 and K8.1 pre-mRNAs each produces three isoforms (alpha, beta, and gamma) of the mRNAs. We have mapped the 5' end of the K8.1 RNA in butyrate-induced KSHV-positive JSC-1 cells to nt 75901 in the KSHV genome and have shown that exon 3 of the K8 pre-mRNA in JSC-1 cells covers most part of the intron 3 defined previously and has three 5'-splice sites (ss), respectively, at nt 75838, 76155, and 76338. Selection of the nt 75838 5'-ss dictates the K8 mRNA production and overwhelms the RNA processing. Alternative selection of other two 5'-ss is feasible and leads to production of two additional bicistronic mRNAs, K8/K8.1alpha and -beta. However, the novel bicistronic K8/K8.1 mRNAs translated a little K8 and no detectable K8.1 proteins in 293 cells. Data suggest that production of the K8/K8.1 mRNAs may be an essential way to control K8 mRNAs, especially K8alpha, to a threshold at RNA processing level.


Adults with early diagnosis of phenylketonuria have higher resting energy expenditure than adults with late diagnosis.
To date, there is a gap regarding resting energy expenditure (REE) in adults with phenylketonuria (PKU), whether PKU type and time of diagnosis interfere with REE, and whether the available predictive equations are valid in this population. To compare the REE of adult subjects with PKU with healthy subjects and secondarily, examine the REE of adults with PKU according to type and time of diagnosis, and check the agreement of commonly used predictive equations of REE. Concordance study with adults with PKU and a comparison group (CG) with healthy adults. Anthropometric and body composition assessments and REE assessment by indirect calorimetry (IC) were performed. The results obtained by IC were compared with predictive equations. Sixty-nine adults were evaluated (PKU: 36; CG: 33). The REE of adults with mild and classic PKU is similar (p>.05) and similar to CG (p>.05). The REE of individuals with early diagnosis is higher (p < .05) than the REE of individuals with late diagnosis. The REE obtained by IC differed (p < .05) from all estimated REE. Late diagnosis of PKU showed lower REE compared to individuals with early diagnosis. The REE of adults with PKU does not differ in relation to the type of PKU, nor does it differ from the CG. Predictive equations overestimate REE.


Final part-word repetitions in school-age children: two case studies.
In contrast to the many published accounts of the disfluent repetition of sounds at the beginnings of words, cases where it is predominantly the final parts of words that are repeated have been reported relatively rarely. With few exceptions, those studies that have been published have described either pre-school children or neurologically impaired subjects. The purpose of this case report was to describe final part-word repetitions in the speech of two school-age boys of normal intelligence with no known neurological lesions. Their speech was recorded during spontaneous conversation, reading, and sentence repetition. The repetitions occurred in all three speaking conditions, although the majority of instances were observed in spontaneous speech, and on both content words and function words. The participants exhibited no apparent awareness of the disfluencies, no abnormal muscle tension, and no accessory behaviours. Each child produced word-final repeated fragments whose phonological structure was highly predictable according to his individual set of rules. The results are discussed in terms of possible motor and cognitive explanations for the disfluencies. As a result of this activity, the participant will be able to: (1) summarize prior research into final part-word repetition; (2) describe the detailed characteristics of final part-word repetitions as displayed by two children of normal intelligence; and (3) discuss ways in which the behaviour might be explained as part of a model of speech production.


Against the reduction of modality to essence.
It is a truth universally acknowledged that a claim of metaphysical modality, in possession of good alethic standing, must be in want of an essentialist foundation. Or at least so say the advocates of the reductive-essence-first view (the REF, for short), according to which all (metaphysical) modality is to be reductively defined in terms of essence. Here, I contest this bit of current wisdom. In particular, I offer two puzzles-one concerning the essences of non-compossible, complementary entities, and a second involving entities whose essences are modally 'loaded'-that together strongly call into question the possibility of reducing modality to essence.


Phosphoryl-related directing groups in rhodium(III) catalysis: a general strategy to diverse P-containing frameworks.
Herein, a rhodium(III)-catalyzed oxidative C-H activation of simple arylphosphonates and phosphonamides with subsequent coupling with alkenes (olefination), internal alkynes (hydroarylation and oxidative cyclization), or simple arenes to give access to diverse P-containing functional frameworks is reported.


Allergic symptoms and sensitisation in adolescents with cows' milk allergy and atopic eczema in infancy.
The association between atopic sensitisation, atopic eczema (AE) and asthma is known, but distinct roles of allergies on long-term health are unestablished. Evaluation of allergic symptoms and sensitisation in adolescents who in infancy had AE and verified cows' milk allergy (CMA) or AE and a negative CMA challenge, and controls. Children with AE, with and without CMA, from a randomised controlled study in 1999-2001 examining the effect of probiotics on AE severity at older than 12 months of age, attended a follow-up visit at age 16 to 18, with age-matched controls. Data came from a questionnaire (ISAAC questionnaire), analysis of serum antigen-specific immunoglobulin Es (IgEs), and clinical evaluation. Group comparisons were carried out (χ2 tests and logistic regression). Fifty-two patients with AE and CMA (AE/CMA+ group), 52 with AE and suspicion of CMA (AE/CMA- group), and 57 controls attended a study visit. IgE-mediated sensitisation was significantly more prevalent in the AE/CMA+ group vs the controls, for horse, cat, dog, egg white and wheat (P < .024 for all). For birch, timothy and mugwort (P < .008 for all), sensitisation was more prevalent in both the AE/CMA+ group and the AE/CMA- group vs controls. On the basis of questionnaire data the AE/CMA + group reported a significantly higher lifetime prevalence of wheezing (64% vs 35% and 32%; P = .001), noninfectious rhinitis (85% vs 62% and 56%; P = .004), and hay fever (77% vs 52% and 33%; P < .001) vs the AE/CMA- group and the control group, respectively. Patients with AE and CMA in infancy, as opposed to patients with AE only, or controls, report more allergic symptoms and exhibit more allergic sensitisation in adolescence. This indicates that CMA in infancy is an independent risk factor of allergic disease in adolescence.


Perforated duodenal ulcer after laparoscopic gastric bypass.
A patient with a history of a laparoscopic gastric bypass presented with a perforated duodenal ulcer. The ulcer was laparoscopically oversewn, and an omentoplasty was performed. Postoperatively, a broad spectrum antibiotic and a proton pump inhibitor were administrated. Several questions arise regarding the diagnosis and treatment of duodenal ulcers after gastric bypass. What are the diagnostic tools to detect a duodenal ulcer, and how should Helicobacter pylori be diagnosed after gastric bypass? The key question is whether the bypassed stomach should be resected as a definitive treatment for complicated duodenal ulcers.


The G-113A polymorphism in CYP1A2 affects the caffeine metabolic ratio in a Chinese population.
This study was designed to better understand genetic variation in the cytochrome P450 (CYP) gene CYP1A2 and its impact on CYP1A2 activity in Chinese subjects. CYP1A2 genetic polymorphisms were screened by direct sequencing in 27 selected Chinese subjects. Plasma 1,7-dimethylxanthine/caffeine ratios 5 hours after a 100-mg caffeine administration, used as an index of CYP1A2 in vivo activity, were determined in 422 healthy subjects. Five single-nucleotide polymorphism markers, including G-860A (CYP1A2*1C), T-3594G, G-3113A, A-163C (CYP1A2*1F), and C5347T (CYP1A2*1B), were selected and genotyped by either polymerase chain reaction-restriction fragment length polymorphism or direct sequencing. Thirteen polymorphisms and 2 linkage disequilibrium blocks with a boundary around -2467 were identified at this locus. The allele frequency for -3860A, -3594G, -3113A, -163C, and 5347T was 0.21, 0.15, 0.10, 0.36, and 0.14, respectively, in the CYP1A2-phenotyped cohort. A significant difference in CYP1A2 activity was observed among genotypes of polymorphism G-3113A (P = .038), and CYP1A2 activity in subjects carrying the AA genotype was lower than that in those carrying the GA (P = .096) and GG genotypes (P = .036): -0.45 +/- 0.05 (mean +/- SD), -0.32 +/- 0.16, and -0.29 +/- 0.16, respectively. Further analysis based on haplotype pairs found a 1.92-fold variation (95% confidence interval, 1.13-2.71) in mean CYP1A2 activity between haplotype pairs 13 and 15, and the difference was significant (-0.19 +/- 0.15 versus -0.45 +/- 0.05, P = .016). As compared with haplotype pair 10, haplotype pairs 9 and 15 and most haplotype pairs heterozygous for the haplotype with an A allele at -3113, including pairs 5, 8, and 12, also showed significantly lower CYP1A2 activity (P = .015, .048, .008, .024, and .014 for pairs 5, 8, 9, 12, and 15, respectively). In addition, haplotype pairs 5, 9, and 12 also showed significantly lower CYP1A2 activity than pair 13 (P = .034, .020, and .037 for pairs 5, 9, and 12, respectively). The G-3113A polymorphism is associated with decreased CYP1A2 activity, haplotype pairs 10 and 13 are responsible for high CYP1A2 activity, and haplotype pairs 5, 8, 9, 12, and 15 are responsible for low CYP1A2 activity in Chinese subjects.


Effect of Hoe 140, a new bradykinin receptor antagonist, on bradykinin- and platelet-activating factor-induced bronchoconstriction and airway microvascular leakage in guinea pig.
We have investigated the effect of a new bradykinin receptor antagonist, Hoe 140 (D-Arg- Hyp3,Thi5,D-Tic7,Oic8]-bradykinin), on bradykinin- and platelet-activating factor (PAF)-induced bronchoconstriction and airway microvascular leakage in anesthetized guinea pigs. Extravasation of Evans blue dye and lung resistance were measured simultaneously. Both i.v. (15 nmol/kg) and inhaled bradykinin (1 mM, 45 breaths) caused a significant increase in lung resistance and leakage of dye at all airway levels. Hoe 140 (100 nmol/kg i.v.) almost completely inhibited these airway responses induced by bradykinin except for dye extravasation in trachea induced by inhaled bradykinin. Inhaled PAF (3 mM, 30 breaths) significantly increased lung resistance and leakage of due at all airway levels, but Hoe 140 had no effect on these responses. Bradykinin-induced bronchoconstriction and airway microvascular leakage are predominantly mediated by activation of B2 receptor, since Hoe 140 is a B2 receptor antagonist. Bradykinin receptor-mediated mechanisms do not play an important role on inhaled PAF-induced bronchoconstriction and microvascular leakage.


Perilunar dislocation of the carpus. A diagnosis still often missed.
A retrospective study of patients with dorsal perilunar dislocation of the carpus (DPD) from seven different centres has been made. There were 91 cases with adequate documentation and radiographs; of these, 61 attended for review between 12 months and 22 years (follow-up average: 42 months). Functional results were assessed by a modified Witvoet score (Witvoet & Allieu, 1973). There were only 33/61 good results. Statistical evaluation disclosed that, whatever the morphology of the initial injury, prognosis had a significant relation to the delay of time between accident and diagnosis (p less than 0.001) to the accuracy of reduction and its maintenance (p less than 0.025) and to the rotation degree of the semilunar bone (p less than 0.001). Authors conclude by saying that there is no problem in DPD itself. The problem exist in the fact that these injuries are still often missed and, subsequently, mismanaged.


Crohn's Disease Disturbs the Immune Properties of Human Adipose-Derived Stem Cells Related to Inflammasome Activation.
Crohn's disease (CD) is characterized by the expansion of mesenteric fat, also known as "creeping fat." We explored the plasticity and immune properties of adipose-derived stem cells (ASCs) in the context of CD as potential key players in the development of creeping fat. Mesenteric CD-derived ASCs presented a more proliferative, inflammatory, invasive, and phagocytic phenotype than equivalent cells from healthy donors, irrespective of the clinical stage. Remarkably, ASCs from the subcutaneous depot of patients with CD also showed an activated immune response that was associated with a reduction in their immunosuppressive properties. The invasive phenotype of mesenteric CD ASCs was governed by an inflammasome-mediated inflammatory state since blocking inflammasome signaling, mainly the secretion of interleukin-1β, reversed this characteristic. Thus, CD alters the biological functions of ASCs as adipocyte precursors, but also their immune properties. Selection of ASCs with the best immunomodulatory properties is advocated for the success of cell-based therapies.


Modelling pollutant emissions in diesel engines, influence of biofuel on pollutant formation.
In order to reduce the harmful effect on the environment, European Union allowed using the biofuel blends as fuel for the internal combustion engines. Experimental studies have been carried on, dealing with the biodiesel influence on the emission concentrations, showing inconclusive results. In this paper numerical model for pollutant prediction in internal combustion engines is presented. It describes the processes leading towards the pollutant emissions, such as spray particles model, fuel disintegration and evaporation model, combustion and the chemical model for pollutant formation. Presented numerical model, implemented in proprietary software FIRE®, is able to capture chemical phenomena and to predict pollutant emission concentration trends. Using the presented model, numerical simulations of the diesel fuelled internal combustion engine have been performed, with the results validated against the experimental data. Additionally, biodiesel has been used as fuel and the levels of pollutant emissions have been compared to the diesel case. Results have shown that the biodiesel blends release lower nitrogen oxide emissions than the engines powered with the regular diesel.


Homologs of aflatoxin biosynthesis genes and sequence of aflR in Aspergillus oryzae and Aspergillus sojae.
The presence, but not expression, of homologs of three structural genes and a regulatory gene necessary for aflatoxin biosynthesis in Aspergillus parasiticus and A. flavus was shown for A. oryzae and A. sojae. Homologs of the regulatory gene aflR were cloned and sequenced from A. oryzae and A. sojae.


mHealth in Cardiovascular Health Care.
Mobile health (mHealth) has been defined as medical and public health practice supported by mobile devices, such as mobile phones, patient monitoring devices and personal digital assistants. Cardiovascular mHealth is, arguably, leading the mHealth space, through innovation, research and implementation, and especially in the areas of prevention, cardiac rehabilitation and education. mHealth includes simple strategies, such as the use of short message service (SMS) or text messages in successful short-term smoking-cessation, weight loss and diabetes management programs. The recent Australian Tobacco, Exercise and Diet Messages (TEXT ME) randomised clinical trial addressed multiple cardiovascular risk factors. mHealth can also involve more complex strategies, such as smart phone applications (apps), global positioning systems (GPS) and Bluetooth technologies. Although many apps could be considered suitable for primary prevention, they are largely unregulated and most are not evidence-based. Some have been well-developed, such as the Food Switch app and an iPhone electrocardiogram (ECG) system. The "explosion" of apps has driven initiatives such as the Mobile Applications Rating Scale (MARS). More recently, the use of sensors to monitor and provide feedback to patients and healthcare providers is being explored. With almost two billion people currently owning a Smartphone, and 50% of adults (globally) predicted to own one by 2018, mHealth provides the prospect of delivering efficient, affordable healthcare services to widespread populations both locally and globally. In particular, it has the potential to reduce socioeconomic disparity and alleviate the burden of cardiovascular disease. There is now a need to rethink traditional health service structures and bioengineering capacity, to ensure mHealth systems are also safe, secure and robust.


Brief report: the Self Harm Questionnaire: a new tool designed to improve identification of self harm in adolescents.
The Self Harm Questionnaire (SHQ) aiming at identification of self-harm in adolescents has been developed and piloted in a sample of 12-17 year olds (n = 100). The adolescents were recruited from both in- and outpatient psychiatric services. Concurrent validity of the SHQ was evaluated by comparing the SHQ results with recorded self harm in the entire clinical records, while the predictive value of the SHQ was assessed by reviewing the clinical records for further episodes of self harm after three months. 71% of the young people endorsed self harm on the SHQ. While 3% showed a false negative result on the SHQ, 20% of the participants disclosed self harm on the SHQ, which was not recorded in their clinical records. At three months follow up there was no statistically significant difference in either sensitivity .95, 95% CI [.72, 1.0] vs. .74, 95% CI [.49, .90] or specificity .35, 95% CI [.25, .46] vs. .51, 95% CI [.39, .62] between the SHQ and the entire clinical record in predicting future episodes of self harm. The SHQ is an important addition to the clinical repertoire designed to improve identification of self harm.


CAMP-dependent protein kinase of sea urchin sperm phosphorylates sperm histone H1 on a single site.
The phosphorylation of sperm specific histone H1 in the sea urchin Strongylocentrotus purpuratus occurs both in vivo and in vitro on a single serine site in the sequence Arg-Lys-Gly-Ser(P)-Ser-Asn-Ala-Arg. This is a preferred sequence for cAMP-dependent protein kinase. The in vitro phosphorylation is completely dependent on cAMP and is inhibited by the peptide protein kinase inhibitor. The protein kinase inhibitor H-8 blocks the in vivo phosphorylation of H1 without damaging motility, the acrosome reaction or the ability of sperm to fuse with and activate eggs.


The experience of patients with complex wounds and the use of NPWT in a home-care setting.
To explore the experience of patients living with complex wounds and the impact of undergoing negative pressure wound therapy (NPWT) as part of their treatment. Qualitative data were collected from eight patients, using semi-structured interviews. A purposive, stratified sampling approach was used to identify participants with a range of wound types, as well as age and gender. Participants were recruited from one primary care trust and were interviewed, following 10 days of treatment with NPWT. The matrix-based 'Framework' approach, from the National Centre for Social Research, was used to structure the analysis. Of the eight participants recruited, five were female and three male, with an age range of 46-77 years. Five overarching categories, with their constituent themes, were identified from the data. The first, developing a wound through crisis, has three themes: a failing body, missed diagnosis and failed professional intervention. The second category, decreased control, has four related themes: poor communication; failed wound healing; poor discharge planning and failure to recover. The third category, increased control, has four themes: understanding what is happening; symptom control; positive professional relationships and returning to health. The fourth category, using NPWT, has two themes: information and understanding of NPWT and expectations and experience of NPWT. The final category sets out participant recommendations about the device and has two themes: device issues and improving professional practice. This study investigated the experience of patients undergoing NPWT for complex wounds in the home setting and reveals a number of the psychosocial effects of using this therapy in this environment. Participants saw NPWT as an active intervention, associated with improved wound healing and symptom control. The participant experience described in this study, and the recommendations they make, provide a valuable resource to inform service improvement programmes and wound research. This study was supported by a research grant from Smith & Nephew Ltd.


miR-101 suppresses colon cancer cell migration through the regulation of EZH2.
colorectal cancer (CRC) is one of the most prevalent types of malignancies worldwide. The incidence of CRC is steadily increasing due to extended life expectancy and aging-related genetic and epigenetic abnormalities. Dysregulation of microRNAs (miRNAs) has been implicated in CRC development. the current study is a basic research study aimed at understanding the molecular mechanism of miR-101 in the pathogenesis of CRC using human samples in vivo and CRC cell lines in vitro. The miRNAs profile from human samples was analyzed by miRNA microarrays and the expression level of single miRNAs were confirmed by qRT-PCR. The validation of the direct target of miR-101 was performed by western blot assay. The cell mobility of CRC was assessed using the Transwell migration assay. downregulation of miR-101 was identified in 39 human CRC tissues and CRC cell lines (HT29 and SW620) when compared to their counterpart control. We further confirmed that the enhancer of zeste homolog 2 (EZH2), a histone methyltransferase, is a direct target of miR-101. Overexpression of EZH2 promoted CRC cell line migration and this effect was inhibited by forcing the expression of miR-101. Thus, we conclude that miR-101 regulated colon cancer cell migration occurs at least partially, though targeting EZH2. our study suggests that miR-101 functions as a tumor suppressor in CRC, and miR-101 may be a potential therapeutic target for CRC treatment.


Using a sociological conceptualization of stigma to explore the social processes of stigma and discrimination of children in street situations in western Kenya.
The leading causes of street involvement worldwide are poverty, family conflict, and abuse. A common misconception is that street involvement is due to delinquency, a belief leading to social exclusion and social inequality for children in street situations (CSS). Exploring community perceptions of CSS and the reproduction of social difference and inequalities can help reduce stigma and discrimination. To explore how stigma and discrimination of CSS was produced and reproduced in specific contexts of culture and power. Social actors including CSS, healthcare providers, children's officers, and police officers in western Kenya. Using a sociological conceptualization of stigma, this qualitative study explored the stigmatization processes that take shape in specific contexts of culture and power. We conducted 41 in-depth interviews and 7 focus group discussions with a total of 100 participants. CSS were often labeled "chokoraa" or garbage picker, a label linked to undesirable characteristics constituting "evils" in society and stereotyped beliefs that they were "delinquents," reinforcing their "otherness" and devalued social status. CSS experienced individual and structural discrimination leading to exclusion from social and economic life. CSS were stigmatized when labeled, set apart, and linked to negative characteristics leading to their experience of status loss and discrimination. CSS's differentness and devalued status served to limit their access to societal resources and deemed them unworthy of equal rights. Interventions involving various social actors are needed to challenge negative stereotypes, reduce stigma, and uphold CSS's human rights.


Evaluation of quality of life among the caregivers of asthmatic children: the new IFABI-R questionnaire.
The health-related quality of life (HRQoL) of asthmatic children and their caregivers is correlated to management of the disease and the presence of certain morbidity indicators. The integral assessment of paediatric asthma must include the evaluation of HRQoL among the caregivers, although existing questionnaires only partially assess the dimensions of this aspect. The present study describes a new questionnaire for evaluating HRQoL among the caregivers, comprising three dimensions (functional, emotional, and socio-occupational). The study involves two phases. A total of 81 patients between 3 and 9 years of age and their caregivers participated in the first phase, involving a qualitative and psychometric study of the preliminary version of the questionnaire (IFABI). A total of 137 patients between 2 and 17 years of age and their caregivers participated in the second phase, in which the revised version of the questionnaire (IFABI-R) was developed and subjected to psychometric evaluation. First phase: The IFABI showed important reliability and internal consistency (Cronbach alpha=0.93), concurrent validity requiring improvement, and a scantly clear internal structure. Second phase: The IFABI-R showed important reliability and internal consistency (Cronbach alpha=0.90), adequate concurrent validity, and a three-dimensional structure whose three factors correspond to the three dimensions of the questionnaire. The good psychometric results obtained with the IFABI-R justify its use in paediatric asthmatic patients. The questionnaire is currently being scaled, and its sensitivity to change is being assessed.


Increased exposure to loading is associated with decreased plantar soft tissue hardness in people with diabetes and neuropathy.
Literature indicates that altered plantar loading in people with diabetes could trigger changes in plantar soft tissue biomechanics which, in turn, could affect the risk for ulceration. To stimulate more research in this area, this study uses in vivo testing to investigate the link between plantar loading and tissue hardness. Tissue hardness and plantar pressure distribution were measured for six plantar areas in 39 people with diabetes and peripheral neuropathy. Spearman correlation analysis revealed that increased pressure time integral at the 1st metatarsal-head region (r = -0.354, n = 39, P = 0.027) or at the heel (r = -0.378, n = 39, P = 0.018) was associated with reduced hardness in the same regions. After accounting for confounding parameters, generalised estimating equations analysis also showed that 10% increase in pressure time integral at the heel was associated with ≈ 1 unit reduction in hardness in the same region. For the first time, this study reveals that people with diabetes and neuropathy who tend to load their feet more heavily also tend to have plantar soft tissues with lower hardness. The observed difference in tissue hardness is likely to affect the tissue's vulnerability to overload injury. More research will be needed to explore the implications of the observed association for the risk of ulceration.


Palmar plating system for Colles' fractures--a preliminary report.
To present our results of the palmar plating system that we developed for the treatment of Colles' fractures. By using the palmar plating system that we developed a consecutive series of 40 acute Colles' fractures were treated surgically. There were 12 men and 28 women with a mean age of 57 years at the time of the injury (range, 25-90 y). All patients had internal fixation using the trans-flexor carpi radialis tendon approach. The system has 3 main features. First, the plate is small in size, being 1.1 mm in thickness and 47 mm in length. Only a 3- to 4-cm skin incision is required for application of the plate. Second, the screw is cannulated and cancellous in type, with a low-profile head. Subchondral screw fixation is achieved both easily and safely by using a guidewire. Third, the plate has a window through which injectable bone cement can be placed. Union was achieved in all patients. The palmar tilt, radial inclination, radial length, and ulnar variance were maintained after surgery. According to the Gartland and Werley rating scale that was modified by Sarmiento there were 12 excellent and 28 good results. There were no extensor tendon injuries that could occur when the dorsal approach was used. This palmar plating system can make fixation of the distal radius easy, safe, and effective in the treatment of unstable Colles' fractures.


Bone marrow-derived mesenchymal stromal cell therapy in a rat model of cavernous nerve injury: Preclinical study for approval.
Although clinical studies using stem cells to treat erectile dysfunction have been performed or are ongoing, there is little consensus on the optimal protocol. We aimed to develop a protocol optimizing human bone marrow-derived mesenchymal stromal cell (hBMSC) therapy in a rat model of cavernous nerve injury. We performed, in order, a dose-finding study, a toxicokinetic study of hBMSCs, and a study to determine the timing and number of cell injections. From the dose-finding study, 1 × 10(6) cells were selected as the dose per hBMSC injection. From the toxicokinetic study, 14 days was selected as the interval between repeat treatments. In the final study, the ratio of maximal intracavernous pressure to mean arterial pressure was significantly lower in the control group than in the sham group (23.4% vs. 55.1%, P <0.001). An immediate single injection of hBMSCs significantly improved erectile function compared with the control group (39.8%, P = 0.035), whereas a delayed single injection showed improvement with a marginal trend (38.1%, P = 0.079). All histomorphometric changes were significantly more improved in the immediate or delayed single injection groups than in the control group. Repeat treatments did not provide any benefit for the recovery of erectile function and histomorphometric changes. Intracavernous injection of 1 × 10(6) hBMSCs results in a recovery of penile erection and histomorphometric changes in a rat model of cavernous nerve injury, even when treatment was delayed until 4 weeks after cavernous nerve injury.


Synthesis of bisindolylmaleimides using a palladium catalyzed cross-coupling reaction.
Bisindolylmaleimides are known to be potent and selective PKC inhibitors. A new synthesis of this class of compound is reported. The key step is a Suzuki cross-coupling reaction using a readily available indolylmaleimide triflate intermediate.


Land-use zoning management to protecting the Regional Key Ecosystem Services: A case study in the city belt along the Chaobai River, China.
There are complex tradeoffs and synergies between ecosystem services. At the regional scale, some ecosystem services play a more critical role in regional ecological security and social development. Therefore, it is necessary to identify key ecosystem services and prioritize their protection and management to ensure their sustainable and effective delivery in a region. This paper proposes a set of research schemes that comprehensively examine the supply, trade-off, and demand of ecosystem services, using the city belt along the Chaobai River in north China as an example for empirical research. Demarcating a 1 km × 1 km area, we first analyze the spatial-temporal supply patterns of Net Primary Productivity (NPP), Water Yield (WY), Soil Retention (SR), Sand Retention (SAR), and Habitat Quality (HQ) from 2010 to 2015 using the CASA, InVEST, USLE, RWEQ and InVEST models, respectively. Thereafter, we define the Regional Key Ecosystem Service (RKES) through regional reality and Spearman correlation analysis among the five ecosystem services. Finally, based on the disparity between the supply and demand of the RKES, we develop a land zoning approach and propose targeted zoning management measures. The results show that WY was the RKES in the study area. The demand for WY did not exceed supply, and the supply and demand were always in a state of mismatch. The stable area with surplus supply and demand of WY was designated as an Ecological Protected Area, a core area to guarantee the RKES. In general, our scheme can be targeted to address the protection of RKES from the perspective of land management. This is of great significance for ensuring regional ecological security and sustainable use of ecosystem services, and could be used to inform the formulation of ecological protection policies.


Subcortical signal alteration of corticospinal tracts. A radiologic manifestation of ARIA: A case report.
Patients with Alzheimer's disease who have been given monoclonal antibodies targeting amyloid-β (Aβ) (eg, gantenerumab, donanemab, lecanemab, and aducanumab) for scientific purposes may have a spectrum of imaging findings known as amyloid-related imaging abnormalities (ARIA), shown on brain magnetic resonance imaging (MRI) scans. These neuroimaging abnormalities are caused by antibody-mediated destruction of accumulated Aβ aggregates in cerebral blood vessels and brain parenchyma. ARIA may demonstrate as brain edema or sulcal effusion (ARIA-E) or as hemosiderin deposits caused by brain parenchymal or pial hemorrhage (ARIA-H). The current study explores 2 cases with interval development of FLAIR hyper signal intensity along the bilateral corticospinal tracts in the motor cortex/precentral gyri after treatment by aducanumab. We believe this manifestation is a subtype of ARIA-A that has not been explored earlier. Our first case was a 72-year-old woman with a history of HTN and kidney transplant (polycystic kidney) who presented with mild cognitive impairment with clinical findings consistent with early Alzheimer's disease. After receiving 3 doses of aducunumab and experiencing cognition improvement, she underwent a brain MRI because of dizziness and vertigo. The brain MRI demonstrated new FLAIR hyper signal intensity in subcortical regions of precentral gyri (motor cortex) symmetrically as well as trace subarachnoid hemorrhage at the vertex compatible with ARIA-E and ARIA-H. Our second case was an 85-year-old woman with a history of small lymphocytic leukemia which was treated 20 years earlier. After orthopedic surgery 2 years ago, she developed dementia with anterograde amnesia. Since then, Aricept and Namenda have been started, but there have been no improvements in her subjective condition. The initial Amyloid PET/MR imaging showed diffuse cerebral Amyloid deposition. After tolerating 6 doses of aducanumab a safety MRI revealed new bilateral symmetric FLAIR hyper signal intensity in the subcortical motor cortex. Results of our study suggest that the subcortical corticospinal tract is another hotspot for ARIA findings. Hence, these regions might be an unknown site for both the action and adverse effects of aducanumab on amyloid plaques with secondary inflammation. In addition, radiologists must take this phenomenon into the account, and be cognizant that the FLAIR hyper signal intensities should not be misinterpreted as motor neuron disease (eg, amyotrophic lateral sclerosis).


Isoelectric focusing of creatine kinase MM isoforms and its application for diagnosis of acute myocardial infarction.
CK MM isoforms (MM 3 having the highest isoelectric point, followed by MM 2, MM 1, and MM X) were measured in 35 patients with acute myocardial infarction (AMI) by isoelectric focusing on agarose gel. Blood samples were analysed every 2 h for the first 12 h, then every 4-8 h until 72 h after AMI. In the first sample, obtained 2.1 h after the onset of chest pain, the ratio of the isoforms MM 3:1 was 0.7 (range 0.2-1.8), equivalent to a normal value. Before the total CK exceeded normal, in 86% of the patients the ratio MM 3:1 rose to 2.2 (range 0.3-3.3). The maximal individual ratio MM 3:1 was 4 (range 0.9-12) after 7 h. It fell below 1 again after 27 h. Thus, the ratio MM 3:1 was useful in the early diagnosis of AMI by enzymatic methods and to estimate the time elapsed since the onset of infarction. Twenty patients with an open infarct vessel (angiographic data after thrombolytic therapy) showed similar peak enzyme activities as ten non-reperfused patients. They differed significantly in the time to the peak activity, mostly for CK MM 3 and CK MB (p less than 0.0005). A higher ratio CK MM 3:1 and a shorter time to the maximum CK MM 3 activity in reperfused patients helps to assess the success of thrombolytic therapy.


Effects of low oxygen levels on the expression and function of transporter OCTN2 in BeWo cells.
Although hypoxia is normal in early pregnancy, low placental oxygen concentrations later in pregnancy are often linked to complications such as pre-eclampsia and intrauterine growth restriction. The effects of low oxygen levels on drug and nutrient uptake via the organic cation transporter OCTN2 has been studied in BeWo cells, an in-vitro model of human trophoblast. BeWo cells were cultured under 20% (control) or 2% O(2) (hypoxia) for 48 h before each experiment. In-vitro hypoxia was also simulated by the addition of CoCl(2) to the cell culture medium. RT-PCR indicated increased transcription of OCTN2 in BeWo cells cultured under hypoxia, but Western blots did not show a corresponding increase in the amount of OCTN2 protein in the hypoxic cells compared with control. Hypoxia resulted in significant reductions in OCTN2-mediated carnitine uptake. Decreased placental transport of carnitine may lead to symptoms of carnitine deficiency in infants from hypoxic pregnancies, whether caused by high altitude, pre-eclampsia or other factors. The OCTN1 substrate ergothioneine reversed the effects of hypoxia on carnitine transport, but identical concentrations of N-acetylcysteine, another water-soluble intracellular antioxidant, did not have the same effect.


Orientation and frequency dependence of backscatter coefficient in normal and pathological breast tissues.
The backscatter coefficient was measured on five groups of normal and pathological breast specimens: (1) as a function of frequency (in the range 4-14 MHz) and (2) at a single fixed frequency (10 MHz), as a function of the angle of incidence between the beam and the specimen (approximately 60 degrees). The results of the study are discussed in relation to the content of cells and collagen fibers in breast tissues. The absolute value of backscatter coefficient is larger in tissues with a prevalence of collagenous fibers in comparison to tissues with only cells. In fibrofatty tissue, the inhomogeneity of the specimens is probably responsible for the highest backscatter value. The power law frequency dependence of the backscatter coefficient is of a diffractive nature in tissues characterized by only cells; in tissues with a strong prevalence of collagenous fibers, the power law frequency dependence increases. Periodicities in the angular patterns have been quantified by the autocorrelation functions for each group of specimens. The results of the study suggest a means for assessing tissue structure in normal and pathological breast tissue.


[Construction of T vectors based on Xcm I recognition site and optimization of PCR fragments for ligation].
To construct T vectors based on Xcm I recognition site and optimize the PCR fragments for its ligation. We firstly cloned the human histone H4 cDNA containing one Xcm I recognition site at both its 5' and 3' end into pCDNA 3.0 vector and then generated T vector with pCDNA 3.0 backbone by cutting the recombinant plasmid with Xcm I. To increase the ligation efficiency, the primers were firstly phosphorylated before DNA fragments amplification and then the PCR products were treated with Taq DNA polymerase and dATP after PCR amplification. Two DNA fragments with the length of 312 bp and 1 329 bp were ligated to it and the ligation mixture was transformed into E. coli DH5α competent cells and the positive rates of the transformants were evaluated by PCR and DNA agarose gel electrophoresis. Our results showed that the T vector produced by our method could ligate to the target DNA fragments with high efficiency. Besides, the phosphorylation state of the primers used for PCR amplification is also an important factor determining the cloning efficiency. What's more, as for longer DNA fragments, retreatment with Taq DNA polymerase and dATP after PCR amplification and purification could improve the ligation efficiency significantly. Our protocol may overcome the dependence on blue/white screening to get positive clones and provide a potent way to generate T vectors and ligate them to the target PCR fragment.


[Pulmonary aspergillosis complicating atypical mycobacterial infection in two patients suffering from chronic obstructive pulmonary disease].
Atypical mycobacteria and Aspergillus are opportunistic organisms responsible for severe pulmonary diseases whose development is encouraged by the presence of chronic obstructive pulmonary disease (COPD) and related immunosuppression. We report the cases of two patients, both alcoholics with emphysematous COPD, who developed chronic pulmonary aspergillosis following atypical mycobacterial infection. Patient 1 developed chronic necrotising aspergillosis several months after the diagnosis of infection with Mycobacterium avium. Patient 2 developed an aspergilloma several weeks after the diagnosis of infection with Mycobacterium xenopi. The association of these two pathologies presents diagnostic and therapeutic problems that are discussed. The development of Aspergillus pulmonary disease may complicate atypical mycobacterial infections and explain a poor response to treatment. Our two case reports suggest that a systematic search should be made for pulmonary aspergillosis during the follow-up of patients with atypical mycobacterial infection.


Antidiuretic hormone in bronchogenic carcinoma.
Immunoactive antidiuretic hormone (ADH) was measured by radioimmunoassay in the plasma, lung tumours and metastatic tumours of nineteen patients with bronchogenic carcinoma. Ten patients had hyponatraemia and carcinoma of the small oat cell type. Plasma ADH measured in nine of these patients ranged from 11--270 pg/ml and was elevated above the normal range (4.6--6.2 pg/ml) in all subjects. ADH-immunoreactive material was detectable in all primary lung tumours (range 9--1080 pg/mg wet weight, n = 7) and metastases (range 5--63 pg/mg wet weight, n = 9) obtained from the hyponatraemic patients. A statistical relationship existed between plasma and tumour ADH concentration in six patients where both measurements were performed. Three patients had small cell carcinomas (two oat cell and one anaplastic) without overt hyponatraemia. ADH-like material was detectable in the lung tumours (18 and 1.1 pg/mg wet weight) and liver metastases (4 and 1.0 pg/mg wet weight) of two patients but not in the third. Four of the remaining patients had squamous cell carcinomas and two had adenocarcinomas. None had hyponatraemia. ADH-like material was undetectable in all lung tumours, metastatic tumours and uninvolved tissue from these patients. ADH extracted from the pituitaries of four patients ranged from 6400--13200 pg/mg wet weight. ADH immunoreactive extracts of six lung tumours and nine metastases (all oat cell) showed the same pattern on elution from a Sephadex G-25 column. A large peak, which made up 65% of the total activity, was eluted in the same position as synthetic arginine vasopressin and contained comparable amounts of immunoreactive and bioactive ADH. Two smaller peaks (8 and 27% of total activity) were eluted in positions of higher molecular weight and contained more immunoreactive than bioactive ADH. In contrast, three of four pituitary extracts showed only a single peak which eluted in the same position as marker vasopressin.


Differential modulation of ATP-induced calcium signalling by A1 and A2 adenosine receptors in cultured cortical astrocytes.
1. Despite the accumulating evidence that under various pathological conditions the extracellular elevation of adenine-based nucleotides and nucleosides plays a key role in the control of astroglial reactivity, how these signalling molecules interact in the regulation of astrocyte function is still largely elusive. 2. The action of the nucleoside adenosine in the modulation of the intracellular calcium signalling ([Ca(2+)](i)) elicited by adenosine 5'-triphosphate (ATP)-induced activation of P2 purinoceptors was investigated on neocortical type-1 astrocytes in primary culture by using single-cell microfluorimetry. 3. Astrocyte challenge with ATP (1-10 microm) elicited biphasic [Ca(2+)](i) responses consisting of an initial peak followed by a sustained elevation. The stable adenosine analogue 2-chloroadenosine (2-ClA) potentiated the transient [Ca(2+)](i) rise induced by activation of metabotropic P2Y receptors. Among the various P1 receptor agonists tested, the nonselective agonist 5'-N-ethylcarboxamidoadenosine (NECA) mimicked the 2-ClA action, whereas the selective A1 R(-) N6-(2-phenylisopropyl)-adenosine (R-PIA), the A2A 2-[4-(2-carboxyethyl)phenethylamino]-5'-N-ethylcarboxamidoadenosine (CGS-21680) and A3 1-deoxy-1-(6-[([3-lodophenyl]methyl)-amino]-9H-purin-9-yl)-N-methyl-beta-d-ribofuranuronamide (IB-MECA) agonists were ineffective. 4. Application of R-PIA>NECA>or=2-ClA depressed the [Ca(2+)](i) plateau reversibly. Moreover, in the presence of R-PIA or 2-ClA, the prolonged [Ca(2+)](i) signal was maintained by application of the A1 antagonist 1,3-diethyl-8-phenylxanthine (DPX). Finally, preincubation of the astrocytes with pertussis toxin abrogated the 2-ClA inhibition of the ATP-elicited sustained [Ca(2+)](i) rise without affecting the transient [Ca(2+)](i) potentiation. 5. Taken together, these findings indicate that stimulation of A1 and A2 adenosine receptors mediates a differential modulation of [Ca(2+)](i) signalling elicited by P2 purinoceptors. Since variations in [Ca(2+)](i) dynamics also affect cell proliferation and differentiation, our data suggest that tuning of the extracellular levels of adenosine may be relevant for the control of astrogliosis mediated by adenine nucleotides.


Medicaid managed care and the unmet need for mental health care among children with special health care needs.
To determine the association between Medicaid managed care pediatric behavioral health programs and unmet need for mental health care among children with special health care needs (CSHCN). The National Survey of CSHCN (2000-2002), using subsets of 4,400 CSHCN with Medicaid and 1,856 CSHCN with Medicaid and emotional problems. Additional state-level sources were used. Multilevel models investigated the association between managed care program type (carve-out, integrated) or fee-for-service (FFS) and reported unmet mental health care need. The National Survey of CSHCN conducted telephone interviews with a sample representative at both the national and state levels. In multivariable models, among CSHCN with only Medicaid, living in states with Medicaid managed care (odds ratio [OR]=1.81; 95 percent confidence interval: 1.04-3.15) or carve-out programs (OR=1.93; 1.01-3.69) were associated with greater reported unmet mental health care need compared with FFS programs. Among CSHCN on Medicaid with emotional problems, the association between managed care and unmet need was stronger (OR=2.48; 1.38-4.45). State Medicaid pediatric behavioral health managed care programs were associated with greater reported unmet mental health care need than FFS programs among CSHCN insured by Medicaid, particularly for those with emotional problems.


Seroepidemiology of hepatitis a in the croatian population.
Hepatitis A virus (HAV) has a worldwide distribution, although this distribution tends to be uneven among geographical regions and population groups. The prevalence of anti-HAV antibodies in the general population varies widely among countries. In Europe, the seroprevalence of HAV is reported to range from 32% to 88%. The aim of this study was to determine the seroprevalence of HAV among the general Croatian population. During a 2-year period (2008-2009), a total of 791 serum samples were tested for the presence of anti-HAV total (IgM+IgG) and anti-HAV IgM antibodies using an automated enzyme-linked fluorescent assay (Mini Vidas; bioMérieux, Marcy l'Etoile, France). The overall anti-HAV seroprevalence was 41.6%. The observed difference in the seroprevalence rates among male and female patients was not statistically significant (44% vs. 39.6%, P = 0.218). A marked increase in anti-HAV seropositivity with age was observed (P < 0.001). The seroprevalence did not differ significantly between participants residing in rural regions (45.3%) and those residing in urban regions (40.6%, P = 0.292). Our results corroborate those of seroprevalence studies in other developed countries. More than half of the Croatian population (59.4%) is susceptible to HAV infection. Older age is an important predictor for being anti-HAV positive.


Effect of cisapride on gastric and esophageal emptying in insulin-dependent diabetes mellitus.
The effects of cisapride on gastric emptying, esophageal emptying, gastrointestinal symptoms, and glycemic control were evaluated in 20 insulin-dependent diabetics who had delayed gastric emptying of the solid or liquid component of a meal, or both. A double-isotope technique was used to measure gastric emptying, and esophageal emptying was measured as the time for a bolus of the solid meal to enter the stomach. On 2 days each patient received cisapride (20 mg) or placebo orally, 60 min before an esophageal and gastric emptying test. A third gastric and esophageal emptying test was performed after each patient had orally taken 10 mg of cisapride or placebo q.i.d. for 4 wk. Single-dose cisapride increased esophageal emptying (p less than 0.01) and both solid and liquid gastric emptying (p less than 0.001). The response to cisapride was most marked in patients with the greatest delay in esophageal and gastric emptying (p less than 0.05). After administration of cisapride for 4 wk, gastric emptying of solid and liquid were faster (p less than 0.001), but esophageal emptying was not significantly different from the placebo test. Upper gastrointestinal symptoms were less after cisapride (p less than 0.05), whereas there was no change on placebo (p greater than 0.2). Plasma glucose and glycosylated hemoglobin concentrations were not different after cisapride compared with placebo. These results indicate that single-dose cisapride increases esophageal emptying in insulin-dependent diabetics and that chronic administration of cisapride is effective in the treatment of diabetic gastroparesis.


Characterization of Minimizers of Aviles-Giga Functionals in Special Domains.
We consider the singularly perturbed problem F ε ( u , Ω ) : = ∫ Ω ε | ∇ 2 u | 2 + ε - 1 | 1 - | ∇ u | 2 | 2 on bounded domains Ω ⊂ R 2 . Under appropriate boundary conditions, we prove that if Ω is an ellipse, then the minimizers of F ε ( · , Ω ) converge to the viscosity solution of the eikonal equation | ∇ u | = 1 as ε → 0 .


Determining the Relationship Between Seizure-Free Days and Other Predictors of Quality of Life in Patients with Dravet Syndrome and Their Carers from FFA Registration Studies.
Dravet syndrome (DS) is a rare, lifelong epileptic encephalopathy characterised by frequent and severe seizures associated with premature mortality. Typically diagnosed in infancy, patients also experience progressive behavioural, motor-function and cognitive decline. Twenty percent of patients do not reach adulthood. Quality of life (QoL) is impaired for both patients and their carers. Reducing convulsive seizure frequency, increasing convulsive seizure-free days (SFDs) and improving patient/carer QoL are primary treatment goals in DS. This study explored the relationship between SFDs and patients' and carers' QoL to inform a cost-utility analysis of fenfluramine (FFA). In FFA registration studies, patients (or their carer proxies) completed the Paediatric QoL inventory (PedsQL). These data were mapped to EuroQol-5 Dimensions Youth version (EQ-5D-Y) to provide patient utilities. Carer utilities were collected using EQ-5D-5L and mapped to EQ-5D-3L to align patient and carer QoL on the same scale. Linear mixed-effects and panel regression models were tested and Hausman tests identified the most appropriate approach for each group. On this basis, a linear mixed-effects regression model was used to examine the relationships between patient EQ-5D-Y and clinically relevant variables (age, frequency of SFDs per 28 days, motor impairments and treatment dose). A linear panel regression model examined the relationship between SFDs and carer QoL. After adjustment for age and underlying comorbidities, the patient regression model showed that SFDs per 28 days was a significant predictor of QoL. Each additional patient-SFD increased utility by 0.005 (p < 0.001). The carer linear panel model also showed that increasing SFDs per 28 days was a significant predictor of improved QoL. Each additional SFD increased carer utility by 0.014 (p < 0.001). This regression framework highlights that SFDs are significantly correlated with both patients' and carers' QoL. Treatment with effective antiseizure medications that increase SFDs directly improves QoL for patients and their carers.


Chemometrics quality assessment of wastewater treatment plant effluents using physicochemical parameters and UV absorption measurements.
Chemometric techniques like Principal Component Analysis (PCA) and Partial Least Squares Regression (PLS) are used to explore, analyze and model relationships among different water quality parameters in wastewater treatment plants (WWTP). Different data sets generated by laboratory analysis and by an automatic multi-parametric monitoring system with a new designed optical device have been investigated for temporal variations on water quality parameters measured in the water influent and effluent of a WWTP over different time scales. The obtained results allowed the discovery of the more important relationships among the monitored parameters and of their cyclic dependence on time (daily, monthly and annual cycles) and on different plant management procedures. This study intended also the modeling and prediction of concentrations of several water components and parameters, especially relevant for water quality assessment, such as Dissolved Organic Matter (DOM), Total Organic Carbon (TOC) nitrate, detergent, and phenol concentrations. PLS models were built to correlate target concentrations of these constituents with UV spectra measured in samples collected at (1) laboratory conditions (in synthetic water mixtures); and at (2) WWTP conditions (in real water samples from the plant). Using synthetic water mixtures, specific wavelengths were selected with the aim to establish simple and reliable prediction models, which gave good relative predictions with errors of around 3-4% for nitrates, detergent and phenols concentrations and of around 15% for the DOM in external validation. In the case of nitrate and TOC concentrations modeling in real water samples from the effluent of the WWTP using the reduced spectral data set, results were also promising with low prediction errors (less than 20%).


Effects of the Surface Densities of Glycoclusters on the Determination of Their IC50 and Kd Value Determination by Using a Microarray.
Pseudomonas aeruginosa (PA) is an opportunistic bacterium involved in 10-30% of nosocomial diseases. It causes severe lung injury to cystic fibrosis patients, often leading to patient death. PA strains are multidrug resistant, thus making the design of new therapeutics a challenge for public health. One promising therapeutic option is to design glycoclusters that target the virulence factor of PA. LecA is a galactose-specific lectin that might be involved in adhesion and biofilm formation by PA. The DNA-directed immobilization (DDI) microarray is a powerful tool for screening and understanding of structure-activity relationships between glycoclusters and lectins. High-throughput and multiplexed analysis of lectin-glycocluster interactions on a DDI microarray allows measurement of IC50 and dissociation constant (Kd ) values with minute amounts of material. In order to study the robustness of the DDI microarray in determination of IC50 and Kd values, the impact of glycocluster surface density was investigated. The data obtained show that measured IC50 values were influenced by glycocluster surface density: as the density of glycoclusters increases, the measured IC50 values increase too. In contrast, the measured Kd values were not affected by glycocluster surface density, provided that the experimental conditions allow interaction between glycocluster and lectin at single-molecule level (no surface cluster effect).


Mortality predictors in liver transplant recipients with recurrent hepatitis C cirrhosis.
Recipients of orthotopic liver transplant for hepatitis C (HCV) invariably develop recurrent disease. The risk factors for death and retransplantation following the development of cirrhosis from HCV are unclear. The aim of this study was to identify predictors of survival in liver transplant recipients who develop cirrhosis from recurrent HCV. We reviewed records of patients who underwent liver transplantation for cirrhosis due to HCV between January 1990 and December 2001. Prognostic factors of patient survival following the development of recurrent cirrhosis were identified through multivariate analysis. During the study period, 511 patients underwent transplantation for HCV cirrhosis. Of these, 65 patients (13%) developed biopsy proven recurrent cirrhosis from HCV; 43 (8%) were relisted for transplantation, and 24 (5%) underwent retransplantation. The overall Kaplan-Meier patient survival rates after the histologic diagnosis of cirrhosis at 1 and 5 years were 66% and 30%, respectively. A multivariate Cox proportional hazards model showed patients with higher last (i.e. at follow-up or prior to retransplantation) International normalized ratio (INR) values (hazard ratios (HR)=2.02, 95% confidence interval 1.26, 3.24, P<0.01) to have an increased risk of death. Our results suggested survival was decreased after the diagnosis of cirrhosis from recurrent HCV. Higher INR was associated with an increased risk of death following the development of cirrhosis.


Bean mild mosaic virus: genome and its translation products.
Viral RNA from purified preparations of bean mild mosaic virus (BMMV) was resolved into two RNAs of mol. wt 1.5 X 10(6) daltons (R1) and 0.76 X 10(6) daltons (R2). Complementary DNA hybridizations, infectivity tests, and in-vitro translation studies established that R2 is a subgenomic RNA derived from R1. In-vitro translation studies showed that the genomic RNA (R1) coded for two major proteins of mol. wt. 40.5 kD and 25.7 kD. Of these, the 40.5 kD protein was shown to be the capsid protein that is translated in vitro from both the R1 and R2 RNAs. Time-course studies on the appearance of in-vitro translation products showed that the 25.7 kD product is the earliest product made when the genomic R1 RNA is translated. This is not one of the products of R2 RNA. Several of the characteristics of BMMV described in this paper show similarities with turnip crinkle virus (TCV) and carnation mottle virus (CarMV).


Continuous noninvasive glucose monitoring; water as a relevant marker of glucose uptake in vivo.
With diabetes set to become the number 3 killer in the Western hemisphere and proportionally growing in other parts of the world, the subject of noninvasive monitoring of glucose dynamics in blood remains a "hot" topic, with the involvement of many groups worldwide. There is a plethora of techniques involved in this academic push, but the so-called multisensor system with an impedance-based core seems to feature increasingly strongly. However, the symmetrical structure of the glucose molecule and its shielding by the smaller dipoles of water would suggest that this option should be less enticing. Yet there is enough phenomenological evidence to suggest that impedance-based methods are truly sensitive to the biophysical effects of glucose variations in the blood. We have been trying to answer this very fundamental conundrum: "Why is impedance or dielectric spectroscopy sensitive to glucose concentration changes in the blood and why can this be done over a very broad frequency band, including microwaves?" The vistas for medical diagnostics are very enticing. There have been a significant number of papers published that look seriously at this problem. In this review, we want to summarize this body of research and the underlying mechanisms and propose a perspective toward utilizing the phenomena. It is our impression that the current world view on the dielectric response of glucose in solution, as outlined below, will support the further evolution and implementation toward practical noninvasive glucose monitoring solutions.


The Role of miR-497/EIF3A Axis in TGFβ1-Induced Epithelial-Mesenchymal Transition and Extracellular Matrix in Rat Alveolar Epithelial Cells and Pulmonary Fibroblasts.
Multi-cause-induced interstitial lung disease, particularly pulmonary fibrosis, is a serious clinical concern. Fibroblasts have been suggested to have a major role, with it recently being revealed that some of these fibroblasts are derived from alveolar epithelial cells through epithelial-mesenchymal transition (EMT). Eukaryotic translation initiation factor 3 subunit A (EIF3A) is a protein that in humans is encoded by the EIF3A gene, and has been suggested to play roles in regulating translation of a subset of mRNAs and in regulating cell cycle progression and cell proliferation. In the present study, we chose a well-known TGFβ1-induced EMT model in alveolar epithelial cells to investigate the functional role of EIF3A. TGFβ1-induced EIF3A expression and EMT process in alveolar epithelial cells, after EIF3A knockdown, the EMT process could be partially reversed. Online tools and luciferase assays showed that miR-497 could inhibit EIF3A expression by directly binding to the 3'UTR of EIF3A. Ectopic miR-497 expression partially reversed TGFβ1-induced EMT in alveolar epithelial cells. In addition, miR-497 could suppress TGFβ1-induced pulmonary fibroblast proliferation and EIF3A, Collagen I and α-SMA protein levels. Taken together, EIF3A could promote TGFβ1-induced EMT in alveolar epithelial cells; miR-497 suppressed TGFβ1-induced EMT in alveolar epithelial cells TGFβ1-induced excessive proliferation and ECM in pulmonary fibroblast through inhibiting EIF3A by targeting. MiR-497/EIF3A axis shows the potential to be effective in the treatment of pulmonary fibrosis. J. Cell. Biochem. 118: 3401-3408, 2017. © 2017 Wiley Periodicals, Inc.


Characteristics of surgically induced astigmatism after standardized microincisional cataract surgery with a superior limbal incision.
To determine (1) if measurements of surgically induced astigmatism (SIA) as measured by keratometry (K) and total keratometry (TK) differ (2) if SIA affects the magnitude and/or meridian of keratometric astigmatism (3) if SIA evolves over time. Tertiary care center. Retrospective data analysis. A swept-source optical coherence tomography biometry dataset (IOLMaster700) consisting of 498 eyes (327 patients) from a tertiary care center was analyzed. For all eyes preoperative and postoperative biometric measurements at 1-month, 3-month, and 6-months postoperative visits were considered for vector analysis of SIA K and SIA TK . Centroids in right and left eyes were 0.26 diopters (D) @5 degrees/0.31 D @1 degree for SIA K and 0.27 D @4 degrees/0.34 D @1 degree for SIA TK . Centroids for difference vectors K-TK in right and left eyes were 0.02 D @ 176 degrees/0.03 D @6 degrees. The mean SIA magnitudes in right and left eyes were 0.48 ± 0.41 D and 0.50 ± 0.37 D for SIA K and 0.53 ± 0.42 D and 0.54 ± 0.40 D for SIA TK . In eyes with ATR astigmatism, an increase in postoperative astigmatism magnitude was more common than a decrease. More than 30% of eyes showed changes in the meridian of more than 15 degrees. Overall, we observed differences in K- and TK-derived SIA, and changes in SIA magnitude over time. For postsurgical interventions, postoperative astigmatism meridian values should be measured to base treatments. Astigmatism magnitude showed a tendency to decrease for steep-meridian incisions and to increase in flat-meridian incisions.


Murine echocardiography: a practical approach for phenotyping genetically manipulated and surgically modeled mice.
There have now been literally hundreds of genetically manipulated mouse models developed during the past decade of cardiac research. Echocardiography is considered an extremely important tool to noninvasively assess and serially follow the phenotype of genetically and surgically altered mice. This review describes in detail the technical considerations, various routinely used methods to assess cardiac function, and some emerging techniques in the assessment of cardiac function in experimental mouse models of cardiac disease.


Onset of action of indacaterol in patients with COPD: comparison with salbutamol and salmeterol-fluticasone.
indacaterol is a novel, inhaled once-daily ultra-long-acting β(2)-agonist for the treatment of chronic obstructive pulmonary disease (COPD). this study compared the onset of action of single doses of indacaterol 150 and 300 μg with salbutamol 200 μg, salmeterol-fluticasone 50/500 μg, and placebo in moderate-to-severe COPD patients. this was a multicenter, randomized, double-blind, placebo-controlled crossover study. The primary variable was forced expiratory volume in one second (FEV(1)) at five minutes postdose. out of 89 patients randomized (mean age 62 years), 86 completed the study. At five minutes postdose, both indacaterol doses were statistically and clinically superior to placebo (P < 0.001), with treatment-placebo differences in FEV(1) of 100 (95% confidence interval [CI] 70-130) mL and 120 (95% CI 90-150) mL for indacaterol 150 and 300 μg, respectively. FEV(1) at five minutes postdose with both indacaterol doses was numerically higher than for salbutamol (10 and 30 mL for indacaterol 150 and 300 μg, respectively) and significantly higher than for salmeterol-fluticasone (50 mL, P = 0.003; 70 mL, P < 0.001, respectively). Moreover, both indacaterol doses showed significantly higher FEV(1) than placebo (P < 0.001) at all postdose time points. The numbers of patients with an FEV(1) increase of at least 12% and 200 mL at five minutes postdose were 16 (18.8%), 24 (27.6%), 20 (23.3%), 8 (9.1%), and 3 (3.4%) for indacaterol 150 and 300 μg, salbutamol 200 μg, salmeterol-fluticasone 50/500 μg, and placebo, respectively. single doses of indacaterol 150 and 300 μg demonstrated a fast onset of action similar to that for salbutamol and faster than that for salmeterol-fluticasone.


Binding of sulfamethazine to β-cyclodextrin and methyl-β-cyclodextrin.
β-cyclodextrin (βCD) and methyl-β-cyclodextrin (MβCD) complexes with sulfamethazine (SMT) were prepared and characterized by different experimental techniques, and the effects of βCD and MβCD on drug solubility were assessed via phase-solubility analysis. The phase-solubility diagram for the drug showed an increase in water solubility, with the following affinity constants calculated: 40.4±0.4 (pH 2.0) and 29.4±0.4 (pH 8.0) M(-1) with βCD and 56±1 (water), 39±3 (pH 2.0) and 39±5 (pH 8.0) M(-1) with MβCD. According to (1)H NMR and 2D NMR spectroscopy, the complexation mode involved the aromatic ring of SMT included in the MβCD cavity. The complexes obtained in solid state by freeze drying were characterized by Fourier transform infrared spectroscopy, scanning electron microscopy, and thermal analysis. The amorphous complexes obtained in this study may be useful in the preparation of pharmaceutical dosage forms of SMT.


Biotechnological perspectives on algae: a viable option for next generation biofuels.
Because of their biofuel producing capabilities, algae (including microalgae and cyanobacteria) are effective and sustainable tools to attain energy security with a growing world population and for reduction of our current reliance on fossil fuels. Algal metabolic and genetic engineering could provide substantial advancements in producing novel and promising strains for the production of alternative biofuels. In this review, we have highlighted biotechnological strategies for microalgae and cyanobacteria that target the improvement of: (1) biosynthesis of biofuel precursors (fatty acid, TAGs, and lipids etc.), (2) carbon-capture ability to accumulate more lipids, and (3) engineering hydrogenases for augmented production of biohydrogen. Other strategies for improving quality and quantity of algal biofuels are also explored.


Identification of immune-related lncRNA panel for predicting immune checkpoint blockade and prognosis in head and neck squamous cell carcinoma.
Immunotherapy is changing head and neck squamous cell carcinoma (HNSCC) treatment pattern. According to the Chinese Society of Clinical Oncology (CSCO) guidelines, immunotherapy has been deemed as first-line recommendation for recurrent/metastatic HNSCC, marking that advanced HNSCC has officially entered the era of immunotherapy. Long non-coding RNAs (lncRNAs) impact every step of cancer immunity. Therefore, reliable immune-lncRNAs able to accurately predict the immune landscape and survival of HNSCC are crucial to clinical management. In the current study, we downloaded the transcriptomic and clinical data of HNSCC from The Cancer Genome Atlas and identified differentially expressed immune-related lncRNAs (DEir-lncRNAs). Further then, Cox and least absolute shrinkage and selection operator (LASSO) regression analyses were performed to identify proper DEir-lncRNAs to construct optimal risk model. Low-risk and high-risk groups were classified based on the optimal cut-off value generated by the areas under curve for receiver operating characteristic curves (AUC), and Kaplan-Meier survival curves were utilized to validate the prediction model. We then evaluated the model based on the clinical factors, immune cell infiltration, and chemotherapeutic and immunotherapeutic efficacy between two groups. In our study, we identified 256 Deir-lncRNAs in HNSCC. A total of 18 Deir-lncRNA pairs (consisting of 35 Deir-lncRNAs) were used to construct a risk model significantly associated with survival of HNSCC. Cox proportional hazard regression analysis confirmed that our risk model could be served as an independent prognostic indicator. Besides, HNSCC patients with low-risk score significantly enriched of CD8+ T cell, and corelated with high chemosensitivity and immunotherapeutic sensitivity. Our risk model could be served as a promising clinical prediction indicator, effective discoursing of the immune cell infiltration of HNSCC patients, and distinguishing patients who could benefit from chemotherapy and immunotherapy.


[Accreditation and digitization - just a load of crap? Systematic risk analysis among medical employees - today and 12 years ago].
The aim of this study was to elucidate whether measures of quality management and digitization have the potential to reduce treatment risks in patients of a surgical clinic. All health professional involved in the treatment of patients were asked to participate in a systematic, process-orientated and anonymous survey to assess the probability of occurrence of 69 treatment risks in stationary patients. The surveys were conducted in 2006 before establishing quality management (QM) and digitization, and recently after various certification activities have been performed and the digitization has been completed. According to the survey respondents, QM measures and digitization have led to a significant reduction of the probability of occurrence of 20 treatment risks, although the number of surgeries performed rose 1.8-fold while the number of employees increased by just 1.2-fold to 1.4-fold. The risk reduction was most pronounced regarding mistaken patient identity errors, while complex process risks like insufficient postoperative aftercare or patient dissatisfaction with ineffective communication remained unchanged. An increase in process risks that may be due to an increased workload can be mitigated by QM and digitization measures. This requires a quality and risk management system that is organized by the administration, supported by responsible and risk-aware employees and not imposed. Health professionals estimate that digitization and QM measures have the potential to reduce patients' treatment risks and help offset the increased workload. In particular, accreditations can help implement and maintain quality management measures.


Expression of insulin-like growth factors at mRNA levels during the metamorphic development of turbot (Scophthalmus maximus).
Insulin-like growth factors I and II (IGF-I and IGF-II) are important regulators of vertebrate growth and development. This study characterized the mRNA expressions of igf-i and igf-ii during turbot (Scophthalmus maximus) metamorphosis to elucidate the possible regulatory role of the IGF system in flatfish metamorphosis. Results showed that the mRNA levels of igf-i significantly increased at the early-metamorphosis stage and then gradually decreased until metamorphosis was completed. By contrast, mRNA levels of igf-ii significantly increased at the pre-metamorphosis stage and then substantially decreased during metamorphosis. Meanwhile, the whole-body thyroxine (T4) levels varied during larval metamorphosis, and the highest value was observed in the climax-metamorphosis. The mRNA levels of igf-i significantly increased and decreased by T4 and thiourea (TU, inhibitor of endogenous thyroid hormone) during metamorphosis, respectively. Conversely, the mRNA levels of igf-ii remained unchanged. Furthermore, TU significantly inhibited the T4-induced mRNA up-regulation of igf-i during metamorphosis. The whole-body thyroxine (T4) levels were significantly increased and decreased by T4 and TU during metamorphosis, respectively. These results suggested that igf-i and igf-ii may play different functional roles in larval development stages, and igf-i may have a crucial function in regulating the early metamorphic development of turbot. These findings may enhance our understanding of the potential roles of the IGF system to control flatfish metamorphosis and contribute to the improvement of broodstock management for larvae.


Five-year cumulative incidence of overweight and obesity, and longitudinal change in body mass index in Japanese workers: The Japan Epidemiology Collaboration on Occupational Health Study.
The present study aimed to estimate cumulative incidence of overweight and obesity and describe 5-year longitudinal changes in body mass index (BMI) in a large occupational cohort in Japan. Participants were 55 229 Japanese employees, who were aged 20-59 years and attended at all subsequent annual health check-ups between 2009 and 2014. Mixed model analysis was performed to examine the effects of age and cohort by gender on BMI change, with age as a random variable. Cumulative incidence of overweight (23.0≤ BMI <27.5 kg/m2 ) and obesity (BMI ≥27.5 kg/m2 ) was calculated. Logistic regression analysis was used to estimate odds ratios for the incidence of overweight and obesity according to age group. The incidence of overweight and obesity was approximately double in men (28.3% and 6.7%, respectively) compared to women (14.3% and 3.9%, respectively).The incidence of obesity decreased with age in men, but did not differ according to age in women (P for trend: .02 and .89, respectively). Among overweight participants, the incidence of obesity was higher in women (18.9%) than men (14.5%) and decreased with advancing age (P for trend: <.001 in men and .003 in women). Mean BMI was higher in men than women in all age groups throughout the period. Younger cohorts tended to have a higher BMI change compared with older cohorts. In this Japanese occupational cohort, transition from overweight to obesity is higher in women than men, and the more recent cohorts had a higher change in mean BMI than the older cohorts.


Robot deployment in long-term care : Case study on using a mobile robot to support physiotherapy.
Healthcare systems in industrialized countries face the challenge of providing care for a growing number of elderly people. Information technology has the possibility of facilitating this process by providing support for nursing staff and improving the well-being of the elderly through a variety of support systems. Little is known about the challenges that arise from the deployment of technology in care settings; however, the integration of technology into care is one of the core determinants of successful support. This article presents the challenges and options associated with the integration of technology into care using the example of a mobile robot to support physiotherapy for elderly people with cognitive impairment in the European project Spatio-Temporal Representations and Activities for Cognitive Control in Long-Term Scenarios (STRANDS). This article presents the technical challenges associated with the introduction of robots in the context of care as well as the perspectives of physiotherapists involved and an overview of information and experiences gained. It is hoped that this will provide useful information for the work of researchers and practitioners wishing to integrate robotic aids into the caregiving process.


The molecular basis of gender disparities in smoking lung cancer patients.
Gender disparities exist in smoking-related lung cancer epidemiology, but the molecular basis has not been explored so far. We aimed at identifying genes with gender-bias expression pattern in smoking lung cancer patients for understanding the molecular basis of gender bias in smokers using meta-analysis of microarray gene expression data. Transcriptome of around 1100 samples from 13 studies were used in the meta-analysis to identify 'Lung Cancer genes specific to Female-Smokers' (LCFS) and 'Lung Cancer genes specific to Male-Smokers' (LCMS). The expression profiles of these genes were validated with an independent microarray report and TCGA-RNA-sequencing data. The molecular interactions, pathway, and other functional annotations were portrayed for the key genes identified. We identified 1159 gender-biased genes in smoking lung cancer patients. Of these, 400 and 474 genes showed differential expression in cancerous compared to normal lung of women (LCFS) and men (LCMS), respectively. While many up-regulated LCFS were involved in 'immune responses' including T-cell activation, leukocyte cell-cell adhesion, the LCMS were mainly involved in 'positive regulation of gene expression', signaling pathways including RAS, VEGF, insulin-receptor signaling, and 'cell cycle'. The strategic-method identified genes, particularly, SNX20, GIMAP6, MTMR2, FAM171B, IDH1, MOBP, FBXO17, LPXN and WIPF1, which were consistently differentially expressed in at least 4 studies, and in agreement with RNA-Seq data. Exploring their functions could be beneficial to the gender-based diagnosis, prognosis, and treatment of lung cancer in smokers. The current meta-analysis supports existing knowledge of sexual-dimorphism of immune responses in cancer.


Human saliva penetration of coronally unsealed obturated root canals.
Studies have shown significant coronal dye and bacterial leakage following exposure of sealed root canals to artificial and natural saliva. The purpose of this study was to determine the time needed for bacteria in natural saliva to contaminate the entire length of root canals obturated by lateral and vertical condensation techniques. Forty root canals were cleaned and shaped using a step-back technique. Thirty root canals were obturated with gutta-percha and root canal sealer using either lateral or vertical condensation techniques. Forty root canals were obturated without a root canal sealer and served as positive controls. After obturation, the coronal 3 mm of five root canals were sealed with sticky wax and served as negative controls. The coronal portions of the filling materials were placed in contact with human saliva and the number of days required for bacteria in saliva to penetrate the entire root canals were determined. No bacterial leakage occurred in the negative control group. Complete bacterial leakage occurred within 2 days in the positive control group. All root canals were recontaminated in less than 30 days. No statistical significant difference was found between the two methods of obturation.


Interaction betwenn family and child/adolescent with hearing deficiency.
to know the family interaction with the hearing impaired child/adolescent. descriptive and exploratory qualitative research developed at a Special School in Southern Brazil. Participants were 10 primary caregivers of deaf children/adolescents between 10 and 19 years old. The collection took place in November 2017, through semi-structured interviews containing questions about the communication process of deaf children/adolescents with their families. The information was analyzed through thematic analysis. The study was submitted and approved by the Ethics Committee under opinion number 2.333.560. as the main theme of the study "Interaction between the family and the child/adolescent with hearing impairment", it addresses two sub-themes: potentialities and weaknesses in the communication of the family with the child/adolescent with hearing impairment and learning in the care of the child/adolescent with hearing impairment. it was identified that the interaction of the deaf with the family and society is impaired by people's lack of knowledge about the deaf community and the Brazilian Sign Language, which raises concern in caregivers who often overprotect the child/adolescent which may limit the full development of their skills and autonomy.


Multiple insecticide resistance mechanisms in urban population of Anopheles coluzzii (Diptera: culicidae) from Lagos, South-West Nigeria.
Malaria is a major public health challenge in Africa with Nigeria accounting for the highest burden of the disease in the world. Vector control has proved to be a highly effective component of malaria control, however, the development and spread of insecticide resistance in major vectors of malaria have been a major challenge. This study assessed resistance mechanisms in Anopheles coluzzii populations from Kosofe, Lagos mainland and Ojo Local Government Areas in Lagos, Nigeria where An. gambiae s.l is resistant to DDT and Permethrin. WHO susceptibility bioassay test was used in determining resistance status of An. coluzzii to discriminating doses of DDT and Permethrin while synergist assay was used to assess the involvement of monooxygenases in resistance development. Sub-species of An. gambiae s.l (An. gambiae and An. coluzzii) were identified using polymerase chain reaction (PCR) and Restriction Fragment Length Polymorphism (PCR-RFLP) while Allele-Specific Polymerase Chain Reaction (AS-PCR) assay was used to detect knockdown mutation (kdr-West; L1014F). Biochemical assays were used in determining the activities of metabolic enzymes. High DDT resistance was recorded in An. coluzzii populations from the three sites. Mortality rate of mosquitoes exposed confirmed Permethrin resistance in Kosofe (50%) and Lagos mainland (48%) but resistance was suspected in Ojo (96%). All specimens tested were confirmed as An. coluzzii with low kdr frequency; 11.6%, 16.4% and 6.7% in Kosofe, Lagos mainland and Ojo respectively. Pre-exposure to synergist (PBO) before exposure to Permethrin led to increased mortality in all populations. Esterase activity was insignificantly overexpressed in Kosofe (p = 0.849) and Lagos mainland (p = 0.229) populations. In contrast, GST activity was significantly lower in populations from Lagos mainland (63.650 ± 9.861; p = 0.007) and Ojo (91.765 ± 4.959; p = 0.042) than Kisumu susceptible strains (120.250 ± 13.972). Monooxygenase activity was higher in Lagos mainland (2.371 ± 0.261) and Ojo (1.361 ± 0.067) populations, albeit significantly in Lagos mainland (p = 0.007) only. Presence of target-site mutation in all populations, increased mortality with pre-exposure to PBO and elevated monooxygenase in Lagos mainland population were confirmed. Multiple resistance mechanisms in some urban populations of An. coluzzii from Lagos, Nigeria calls for appropriate resistance management strategies.


Cultured vestibular ganglion neurons demonstrate latent HSV1 reactivation.
Vestibular neuritis is a common cause of both acute and chronic vestibular dysfunction. Multiple pathologies have been hypothesized to be the causative agent of vestibular neuritis; however, whether herpes simplex type I (HSV1) reactivation occurs within the vestibular ganglion has not been demonstrated previously by experimental evidence. We developed an in vitro system to study HSV1 infection of vestibular ganglion neurons (VGNs) using a cell culture model system. basic science study. Lytic infection of cultured rat VGNs was observed following low viral multiplicity of infection (MOI). Inclusion of acyclovir suppressed lytic replication and allowed latency to be established. Upon removal of acyclovir, latent infection was confirmed with reverse-transcription polymerase chain reaction and by RNA fluorescent in situ hybridization for the latency-associated transcript (LAT). A total of 29% cells in latently infected cultures were LAT positive. The lytic ICP27 transcript was not detected by reverse-transcription polymerase chain reaction (RT-PCR). Reactivation of HSV1 occurred at a high frequency in latently infected cultures following treatment with trichostatin A (TSA), a histone deactylase inhibitor. VGNs can be both lytically and latently infected with HSV1. Furthermore, latently infected VGNs can be induced to reactivate using TSA. This demonstrates that reactivation of latent HSV1 infection in the vestibular ganglion can occur in a cell culture model, and suggests that reactivation of HSV1 infection a plausible etiologic mechanism of vestibular neuritis.


New (68)Ga-PhenA bisphosphonates as potential bone imaging agents.
In vivo positron emission tomography (PET) imaging of the bone using [(68)Ga]bisphosphonates may be a valuable tool for cancer diagnosis and monitoring therapeutic treatment. We have developed new [(68)Ga]bisphosphonates based on the chelating group, AAZTA (6-[bis(hydroxycarbonyl-methyl)amino]-1,4-bis(hydroxycarbonyl methyl)-6-methylperhydro-1,4-diazepine). Phenoxy derivative of AAZTA (2,2'-(6-(bis(carboxymethyl)amino)-6-((4-(2-carboxyethyl)phenoxy)methyl)-1,4-diazepane-1,4-diyl)diacetic acid), PhenA, 2, containing a bisphosphonate group (PhenA-BPAMD, 3, and PhenA-HBP, 4) was prepared. Labeling of these chelating agents with (68)Ga was evaluated. The ligands reacted rapidly in a sodium acetate buffer with [(68)Ga]GaCl3 eluted from a commercially available (68)Ge/(68)Ga generator (pH4, >95% labeling at room temperature in 5min) to form [(68)Ga]PhenA-BPAMD, 3, and [(68)Ga]PhenA-HBP, 4. The improved labeling condition negates the need for further purification. The (68)Ga bisphosphonate biodistribution and autoradiography of bone sections in normal mice after an iv injection showed excellent bone uptake. New (68)Ga labeled bisphosphonates may be useful as in vivo bone imaging agents in conjunction with positron emission tomography (PET).


Distinguish metabolic profiles and defense enzymes in Alternaria leaf blight resistant and susceptible genotypes of groundnut.
Alternaria leaf blight is major fungal disease of summer groundnut, causes significant loss of haulm and pod yield. Aims of this study were to understand the role of metabolites and phenylpropanoid related enzymes in Alternaria leaf blight resistance and to find out metabolic marker for disease resistance. Alternaria leaf blight resistant (GPBD4 and CS186) and susceptible genotypes (GG2 and TPG41) of groundnut were grown in pots during rabi-summer 2015. Groundnut plants were infected with Alternaria alternata (Fr.) Keissler at 40 days after sowing. 5 days after infection, upper second leaves were collected from both control and infected plants for analysis. A total of 67 metabolites comprising sugars, sugar alcohols, amino acids, organic acids, fatty acids, sterols and phenolic were identified using gas chromatography-mass spectrometry (non-targeted metabolomics). Constitutive levels of alpha-d-galactoside, d-mannitol, d-erythropentitol, glycine, and hexadecanoic acid were observed higher in resistant genotypes compared to susceptible genotypes. Moreover, arabinofuranose, cinnamic acid, 2-butendioic acid, and linoleic acid were observed only in resistant genotypes at both control and infected stage. In susceptible genotypes myo-inositol, glucose and fructose content was increased after infection with pathogen while decreased in resistant genotypes. Resistant genotypes had higher constitutive level of cinnamic and salicylic acid compared to susceptible genotypes. Non-infected leaves of resistant genotypes also had higher activities of phenylalanine ammonia lyase and tyrosine ammonia lyase activities. Our results suggest that metabolites specifically present in resistant genotypes impart defense mechanism against Alternaria pathogen and can be used as bio-marker for screening of germplasm.


Selection of phage-display library peptides recognizing ethanol targets on proteins.
There is a forthcoming link between chronic alcohol consumption and proteins covalently modified by ethanol metabolites and their antibodies. To identify sensitive probes of protein-ethanol conjugates, we screened for the ethanol-altered protein domains with a phage-display combinatorial peptide library. In principle, recognition of the epitopes by the library peptides occurs through protein-protein interactions. A general screening, M13-based library with 10(9) random sequences of linear heptameric peptides was used. The peptides were displayed in five copies each, as fusion proteins with phage's minor coat protein III. They were located on one end of the surface of the phage particles. The targets were a model protein, streptavidin, and protein-ethanol conjugates (hydroxyethyl radical- or acetaldehyde-modified bovine serum albumin). They were either immobilized on a surface by direct coating or affinity captured on floating beads. An enriched library of phages with the tightest peptide binders for each target was selected and amplified in a multiple-cycle biopanning in vitro procedure. Binders were characterized by DNA sequencing of the corresponding phages and by counter-screening with positive and negative targets in either an enzyme-linked immunosorbent assay or plaque assay. We obtained the HPQ motif for streptavidin and two unique subsets of peptides that recognized each ethanol target with a selectivity of two orders of magnitude above the carrier protein and controls. The application of biopanning processes, coupled with phage-display peptide libraries on biological fluids and tissues, could provide a systematic mapping of protein--ethanol conjugates and supply a means for early diagnosis and prognosis of chronic alcohol consumption in human beings.


Functional importance of the Gly cluster in transmembrane helix 2 of the Bordetella pertussis CyaA-hemolysin: Implications for toxin oligomerization and pore formation.
Adenylate cyclase-hemolysin (CyaA) is a major virulence factor of Bordetella pertussis causing whooping cough in humans. We previously showed that two transmembrane helices (α2 and α3) in the hemolysin domain (CyaA-Hly) are crucially involved in hemolytic activity. Here, PCR-based substitutions were employed to investigate a potential involvement in hemolysis of a series of four Gly residues (Gly(530), Gly(533), Gly(537) and Gly(544)) which map onto one face of a helical wheel plot of pore-lining helix 2. All CyaA-Hly mutant toxins were over-expressed in Escherichia coli as 126-kDa soluble proteins at levels comparable to the wild-type toxin. A drastic reduction in hemolytic activity against sheep erythrocytes was observed for three CyaA-Hly mutants, i.e. G530A, G533A and G537A, but not G544A, suggesting a functional importance of the Gly(530)_Gly(533)_Gly(537) cluster. A homology-based structure of the α2-loop-α3 hairpin revealed that this crucial Gly cluster arranged as a GXXGXXXG motif is conceivably involved in helix-helix association. Furthermore, a plausible pore model comprising three α2-loop-α3 hairpins implicated that Gly(530)XXGly(533)XXXGly(537) could function as an important framework for toxin oligomerization. Altogether, our present data signify for the first time that the Gly(530)_Gly(533)_Gly(537) cluster in transmembrane helix 2 serves as a crucial constituent of the CyaA-Hly trimeric pore structure.


Longitudinal study of Lyme borreliosis in a high risk population in Switzerland.
Orienteers from all parts of Switzerland (n = 416) were included in a longitudinal study for Lyme borreliosis. In spring 1986, the seroprevalence was 28.1%. At the beginning of the study, 84.3% of orienteers reported a history of tick bite, and 3.8% reported a past history of Lyme borreliosis. During the first (spring 1986-autumn 1986), second (autumn 1986-spring 1987) and third (spring 1987-autumn 1987) period, rates of seroconversion were 0.6%, 2.7% and 2.1% respectively. During the first and second period, clinical incidence were 1.0% and 0.25% respectively. No active Lyme borreliosis was detected during the third period. Among orienteers who seroconverted during the study (n = 16), only two developed clinical symptoms. Hence, Borrelia burgdorferi infection is often asymptomatic.


Evolutionary conservation and function of the human embryonic stem cell specific miR-302/367 cluster.
miRNA clusters define a group of related miRNAs closely localized in the genome with an evolution that remains poorly understood. The miR-302/367 cluster represents a single polycistronic transcript that produces five precursor miRNAs. The cluster is highly expressed and essential for maintenance of human embryonic stem cells. We found the cluster to be highly conserved and present in most mammals. In primates, seed sequence and miRNA structure are conserved, but inter-precursor sequences are evolving. Insertions of new miRNAs, deletions of individual miRNAs, and a cluster duplication observed in different species suggest an actively evolving cluster. Core transcriptional machinery consisting of NANOG and OCT-4 transcription factors that define stem cells are present upstream of the miR-302/367 cluster. Interestingly, we found the miR-302/367 cluster flanking region to be enriched as a target site of other miRNAs suggesting a mechanism for feedback control. Analysis of miR-302 and miR-367 targets demonstrated concordance of gene set enrichment groups at high gene ontology levels. This cluster also expresses isomiRs providing another means of establishing sequence diversity. Finally, using three different kidney tumor datasets, we observed consistent expression of miR-302 family members in normal tissue while adjacent tumor tissue showed a significant lack of expression. Clustering expression levels of miR-302 validated target genes showed a significant correlation between miR-302/367 cluster miRNAs and a subset of validated gene targets in healthy and adjacent tumor tissues. Taken together, our data show a highly conserved and still evolving miRNA cluster that may have additional unrecognized functions.


Changes in soil characteristics and C dynamics after mangrove clearing (Vietnam).
Of the blue carbon sinks, mangroves have one of the highest organic matter (OM) storage capacities in their soil due to low mineralization processes resulting from waterlogging. However, mangroves are disappearing worldwide because of demographic increases. In addition to the loss of CO2 fixation, mangrove clearing can strongly affect soil characteristics and C storage. The objectives of the present study were to quantify the evolution of soil quality, carbon stocks and carbon fluxes after mangrove clearing. Sediment cores to assess physico-chemical properties were collected and in situ CO2 fluxes were measured at the soil-air interface in a mangrove of Northern Vietnam. We compared a Kandelia candel mangrove forest with a nearby zone that had been cleared two years before the study. Significant decrease of clay content and an increase in bulk density for the upper 35cm in the cleared zone were observed. Soil organic carbon (OC) content in the upper 35cm decreased by >65% two years after clearing. The quantity and the quality of the carbon changed, with lower carbon to nitrogen ratios, indicating a more decomposed OM, a higher content of dissolved organic carbon, and a higher content of inorganic carbon (three times higher). This highlights the efficiency of mineralization processes following clearing. Due to the rapid decrease in the soil carbon content, CO2 fluxes at sediment interface were >50% lower in the cleared zone. Taking into account carbonate precipitation after OC mineralization, the mangrove soil lost ~10MgOCha-1yr-1 mostly as CO2 to the atmosphere and possibly as dissolved forms towards adjacent ecosystems. The impacts on the carbon cycle of mangrove clearing as shown by the switch from a C sink to a C source highlight the importance of maintaining these ecosystems, particularly in a context of climate change.