Stereospecificity in strigolactone biosynthesis and perception.
Plants produce strigolactones with different structures and different stereospecificities which provides the potential for diversity and flexibility of function. Strigolactones (SLs) typically comprise a tricyclic ABC ring system linked through an enol-ether bridge to a butenolide D-ring. The stereochemistry of the butenolide ring is conserved but two alternative configurations of the B-C ring junction leads to two families of SLs, exemplified by strigol and orobanchol. Further modifications lead to production of many different strigolactones within each family. The D-ring structure is established by a carotenoid cleavage dioxygenase producing a single stereoisomer of carlactone, the likely precursor of all SLs. Subsequent oxidation involves cytochrome P450 enzymes of the MAX1 family. In rice, MAX1 enzymes act stereospecifically to produce 4-deoxyorobanchol and orobanchol. Strigol- and orobanchol-type SLs have different activities in the control of seed germination and shoot branching, depending on plant species. This can partly be explained by different stereospecificity of SL receptors which includes the KAI2/HTL protein family in parasitic plants and the D14 protein functioning in shoot development. Many studies use chemically synthesised SL analogues such as GR24 which is prepared as a racemic mixture of two stereoisomers, one with the same stereo-configuration as strigol, and the other its enantiomer, which does not correspond to any known SL. In Arabidopsis, these two stereoisomers are preferentially perceived by AtD14 and KAI2, respectively, which activate different developmental pathways. Thus caution should be exercised in the use of SL racemic mixtures, while conversely the use of specific stereoisomers can provide powerful tools and yield critical information about receptors and signalling pathways in operation.


A comparison of paediatric dentists' and general dental practitioners' care patterns in paediatric dental care.
The aim of this study was to compare the care patterns of paediatric dentists and general dentists in the dental treatment of children in the Netherlands. A case control study was completed based on the financial records of one of the largest Dutch health insurance companies. After medical ethical approval the dental records from 2004, 2005 and 2006 of 16 paediatric dentists were used and compared with the records of general practitioners with the same number of insured paediatric patients from the same age and urbanisation level. Preventive, diagnostic and restorative care for four consecutive age groups (<6, 6-8, 9-11, 12-17 yrs) were used as independent variables. Differences between the dentists and the age groups were tested with the Independent-Sample t-Test and ratios were calculated. Compared with general dentists, paediatric dentists use statistically significant more often rubber-dam (p=0.009) and did more preventive treatments (p<0.001) in children up to aged 11 yrs, more extractions (p<0.001), took more radiographs (p=0.027) and used local analgesia more often (p=0.002) in children until aged 8 yrs and performed more restorations (p=0.02) in children up to 6 yrs of age. There was no significant difference in the care pattern of the dentists for the oldest age group (12-17 yrs). The care-index for paediatric dentists and general dentists from this research was comparable with Dutch epidemiological studies. Compared with general dentists, paediatric dentists have a more extensive treatment approach when treating children. In the youngest age groups the differences are the most pronounced. Further studies are needed to clarify whether the cause is the needs of the patient or an attitude of supervised neglect by the general dentists.


Provenance Solutions for Medical Research in Heterogeneous IT-Infrastructure: An Implementation Roadmap.
Research data generated in large projects raise challenges about not only data analytics but also data quality assessments and data governance. The provenance of a data set - that is the history of data sets - holds information relevant to technicians and non-technicians and is able to answer questions regarding data quality, transparency, and more. We propose an implementation roadmap to extract, store, and utilize provenance records in order to make provenance available to data analysts, research subjects, privacy officers, and machines (machine readability). Each aspect is tackled separately, resulting in the implementation of a provenance toolbox. We aim to do so within the context of HiGHmed, a research consortium established within the medical informatics initiative in Germany. In this testbed of federated IT-infrastructures, the toolbox shall assist each stakeholder in answering domain-specific and domain-agnostic questions regarding the provenance of data sets. This way, we will improve data re-use, transparency, and reproducibility.


How sailfish use their bills to capture schooling prey.
The istiophorid family of billfishes is characterized by an extended rostrum or 'bill'. While various functions (e.g. foraging and hydrodynamic benefits) have been proposed for this structure, until now no study has directly investigated the mechanisms by which billfishes use their rostrum to feed on prey. Here, we present the first unequivocal evidence of how the bill is used by Atlantic sailfish (Istiophorus albicans) to attack schooling sardines in the open ocean. Using high-speed video-analysis, we show that (i) sailfish manage to insert their bill into sardine schools without eliciting an evasive response and (ii) subsequently use their bill to either tap on individual prey targets or to slash through the school with powerful lateral motions characterized by one of the highest accelerations ever recorded in an aquatic vertebrate. Our results demonstrate that the combination of stealth and rapid motion make the sailfish bill an extremely effective feeding adaptation for capturing schooling prey.


Design and synthesis of novel 1,3,5-triphenyl pyrazolines as potential anti-inflammatory agents through allosteric inhibition of protein kinase Czeta (PKCζ).
Much light has been shed on the vital role of protein kinase Czeta (PKCζ) in NF-κB activation and the potential use of PKCζ inhibitors as anti-inflammatory agents. We previously reported a series of 1,3,5-trisubstituted pyrazolines as potent and selective allosteric inhibitors of PKCζ; in that series of compounds, the phenolic OH at the 5-phenyl was essential for binding to the PKCζ PIF pocket. In the present study, we surprisingly found that replacing it by a halogen and at the same time moving the OH to the 3-phenyl still resulted in active compounds. An extension of this class of compounds with a new focused library is presented herein, where the phenolic OH at the 5-phenyl, which was reported to be an irreplaceable feature for activity, was moved to the 3-phenyl and replaced by halogen. The new set of compounds maintained the same level of potency against PKCζ and selectivity against PKC isoforms, and showed reduced potency against the PIF pocket mutant PKCζ[Val297Leu]. Of note, the repositioning of the key functional groups resulted in a marked enhancement of cellular potency. One of the most potent new PKCζ inhibitors, 2h, was able to suppress NO production in RAW 264.7 macrophage cells with 8 times higher efficacy than the previous series, and inhibited the NF-κB transcriptional activity in U937 cells with a sub-micromolar IC50.


Exploring the Relations of Morphological Awareness with Phonological Awareness and Vocabulary: The Case of the Greek Language.
Although relations between morphological awareness, phonological awareness, and vocabulary have been widely observed, questions remain about their precise associations. The purpose of the present study was to explore the relations of morphological awareness with two highly related linguistic skills (phonological awareness and vocabulary) in a transparent orthography with rich morphology. The study sample consisted of 121 (58 males, Mean age = 93.94 months, SD = 3.32) 2nd grade Greek-speaking children. Confirmatory factor analysis revealed that the three-factor model provided the best fit to the data, indicating that although morphological awareness, phonological awareness, and vocabulary are highly correlated, they represent distinct linguistic constructs. In addition, hierarchical regression analysis was used to examine the bidirectional effects between the three linguistic skills. Results revealed that both phonological awareness and vocabulary significantly contributed to morphological awareness, with phonological awareness having a stronger effect. Conversely, morphological awareness significantly affected both phonological awareness and vocabulary. The effect size from phonological awareness and vocabulary to morphological awareness was similar to the effect size reported from morphological awareness to phonological awareness and vocabulary. These results suggest that morphological awareness is highly associated with phonological awareness and vocabulary, being though a distinct skill. In addition, it seems that these linguistic skills have bidirectional effects with each other in first grades.


Therapeutic strategy for treating aortoesophageal fistulas.
The development fistulas between the thoracic aorta and the esophagus are highly fatal conditions. We aimed to identify a therapeutic strategy for treating aortoesophageal fistula (AEF) in this study, by investigating all AEF cases presented in this special symposium at the 65th Annual Scientific Meeting of the Japanese Association for Thoracic Surgery. Forty-seven AEF patients were included in this study. The survivors and nonsurvivors at six and 18 months after diagnosis of AEF were classified into "Group A6", "Group D6", "Group A18", and "Group D18", respectively. Comparisons between Group A6 and Group D6 and between Group A18 and Group D18 were made with regard to therapeutic strategy. Twenty-two (46.8 %) and 33 (70.3 %) of the 47 patients died within 6 and 18 months, respectively. The patients treated with omentum wrapping (p = 0.0052), esophagectomy (p = 0.0269) and a graft replacement strategy for the aorta (p = 0.002) were more frequently included in Group A6. The patients with the omentum wrapping (p = 0.0174) and esophagectomy (p = 0.0203) and graft replacement were more significantly included in Group A18. The results of the multivariate analysis indicated that the mortality rate at 6 and 18 months after diagnosis was significantly correlated with graft replacement (p = 0.0188) and esophagectomy (p = 0.0257), respectively. There were significant differences in the actuarial survival curves in patients who had omentum wrapping, graft replacement, and esophagectomy compared to patients who did not have these 3 therapeutic procedures. The use of thoracic endovascular aortic repair alone for AEF should not be considered a definitive surgery. In contrast, esophagectomy, open surgery with aortic replacement using prostheses and homografts and greater omentum wrapping significantly improve the mid-term survival of AEF.


ADVERSE DRUG REACTIONS: FACTORS AND ROLE OF PHARMACIST IN THEIR PREVENTION.
Adverse drug reactions (ADRs) are the undesirable effects of drugs even when administered in daily normal doses. It has been seen that ADRs may arise even after single administration of drug(s), however, long term therapies are more prone to ADRs. Currently it has become a burning issue all around the world and connected with financial expansion owing to hospitalization. Pharmacists are the custodians of drugs especially clinical pharmacists play a vital role in preventing the risks of ADRs. This review deals with the factors leading to possible ADRs, its prevention and the role of pharmacist in management of ADRs.


Occupational diseases in South African mines--a neglected epidemic?
A major difficulty in assessing incidence and prevalence of occupational diseases among the workforce employed on South African mines is the lack of epidemiological data. Published and unpublished data were collated and analysed for the period 1915-1988 in order to provide insights into occupational disease trends. Using proxies for incidence rates, it was demonstrated that the rates of silicosis and tuberculosis were likely to be high. In the case of silicosis among black miners, disease was contracted in a relatively short period on average. The stabilisation of the black workforce on the mines, which started in 1975 and continues, is an important factor likely to lead to an increase in occupational disease rates. Recommendations for improved data reporting and research are made, in particular the need to identify occupational groups at high risk and the calculation of prevalences.


Oral hairy leukoplakia which occurred as a presenting sign of acute myeloid leukemia in a child.
Oral hairy leukoplakia (OHL) is caused by the reactivation of a previous Epstein-Barr virus (EBV) infection in the epithelium of the tongue. Most lesions are characterized by corrugated whitish patches on the lateral border of the tongue. It is frequently associated with AIDS, but cases in patients with other immunosuppressed states have also been reported. In leukemia patients, OHL is rarely encountered, and appears only after chemotherapy. We report a case of OHL which occurred as a presenting sign of acute myeloid leukemia (AML) in a previously healthy 15-year-old child. A 15-year-old boy presented with a whitish patch on the left lateral border of the tongue. The biopsy specimen revealed papillomatosis, hyperkeratosis, acanthosis and ballooning degeneration in the stratum spinosum. The patient was EBV seropositive, and PCR analysis of EBV DNA in the lesional tissue was positive. After the diagnosis of OHL in dermatologic department, the patient was referred to pediatrics due to the abnormal peripheral blood smear, and was diagnosed with AML.


Current management of Duchenne muscular dystrophy in the Middle East: expert report.
Aim: Duchenne muscular dystrophy (DMD) is a severe and rare X-linked neuromuscular childhood disorder that results in functional decline, loss of ambulation and early death due to cardiac or respiratory failure. The objective of this paper is to address different aspects of the current management of DMD in the Middle East, north Africa (MENA) region, and to gather experts' recommendations on how to optimally diagnose and treat patients suffering from this disease. Methods: A group of experts (neuromuscular medicine, neuropediatricians and geneticists) convened to discuss the diagnosis and management of DMD in the MENA region. A list of practical statements was prepared by the chair of the meeting to guide the discussions around critical aspects relating to the current and future management of DMD. Results & conclusion: Ideally, DMD management should be a multidisciplinary approach. Nevertheless, few tertiary care hospitals in the region are currently able to provide the full spectrum of medical expertise and services needed by DMD patients. Clinical practice in the region remains heterogeneous. Specific guidelines for diagnosis and treatment are needed in the MENA region to improve outcomes. Disease awareness among the general public and the medical community is lacking. Now that mutation-specific therapies are being developed and more widely studied, general education programs regarding early signs and symptoms, a standardized referral and diagnosis pathway, patient registries and support groups will significantly improve the management of the disease.


[Access barriers to occupational risk coverage in service provision contracts in the public health sector in Bogotá].
Objective To identify access barriers to the General System for Occupational Risks (SGRL in Spanish) in a group of independent professionals (contracted with service provision contraction) in the public health sector in Bogotá. Methodology Qualitative research (narrative) through interviews of four independent professionals of the Secretaría Distrital de Salud (District Health Secretariat), 4 independent professionals of Tunjuelito Hospital and 3 experts in the field of occupational risks. The content analysis paradigm was used for information analysis. Results The two institutions of the public health sector in Bogotá consulted have implemented strategies that facilitate the access to the SGRL for independent professionals. However, the interviewed workers and experts identified some access barriers membership for independent workers in this occupational risk system; among these barriers are the lack of a culture of occupation risk prevention, lack of knowledge about laws and the utility of the system as well as the non-conception of the SGRL as a fundamental part of the social security system of the country. Conclusion It is necessary for the SGRL to be recognized as an integral and fundamental part of the country's social security. This depends on a cultural construction of the importance of an occupational risk system among the state organizations, employers and employees.


Worried together: a qualitative study of shared anxiety in patients with metastatic non-small cell lung cancer and their family caregivers.
Anxiety is prevalent, distressing, and understudied among patients with advanced lung cancer and their family caregivers. Preliminary evidence suggests that anxiety is not only present in both patients and caregivers but shared by the dyad. Few studies have examined the nature of shared anxiety and its impact on patient-caregiver dyads. This study was developed to identify shared causes and manifestations of anxiety experienced by patients with stage IV non-small cell lung cancer (NSCLC) and their primary caregivers. Data were collected through in-depth semi-structured interviews with ten matched patient-caregiver dyads and one unmatched patient (N = 21) recruited from two comprehensive cancer care centers. Using grounded theory, eight themes emerged characterizing shared causes and manifestations of anxiety: (1) uncertainty, (2) loss and impending loss, (3) changing roles, (4) conflict outside the dyad, (5) finances, (6) physical symptoms, (7) fears of decline and dying, and (8) life after the patient's passing. All themes were shared by patients and caregivers. Implications for future research include the development and evaluation of interventions to reduce anxiety in cancer patient-caregiver dyads.


The effect of tibolone treatment on apolipoproteins and lipoprotein (a) concentrations in postmenopausal women: A meta-analysis of randomized controlled trials.
Tibolone is a synthetic steroid with estrogenic, androgenic and progestogenic properties that is used as hormone replacement therapy (HRT) in postmenopausal women. Treatment with tibolone has been demonstrated to lead to changes of the lipid profile, including alterations in lipoprotein (a) and apolipoprotein levels. Hence, we conducted the present meta-analysis of randomized controlled trials (RCTs) to assess the effect of tibolone treatment on apolipoproteins and lipoprotein (a) values in postmenopausal women. Several databases (Cochrane Library, PubMed/Medline, Scopus, and Google Scholar) were searched for English-language manuscripts published up to September 2023 that scrutinized the effects of tibolone administration on apolipoprotein A-I (ApoA-I), apolipoprotein A-II (ApoA-II), apolipoprotein B (ApoB), and lipoprotein (a) in postmenopausal women. The results were reported as the weighted mean difference (WMD) with a 95% confidence interval (CI), generated using a random-effects model. Finally, 12 publications with 13 RCT arms were included in the current meta-analysis. The overall results from the random-effects model demonstrated a notable reduction in ApoA-I (n = 9 RCT arms, WMD: -34.96 mg/dL, 95 % CI: -42.44, -27.48, P < 0.001) and lipoprotein (a) (n = 12 RCT arms, WMD: -7.49 mg/dl, 95 % CI: -12.17, -2.81, P = 0.002) after tibolone administration in postmenopausal women. However, treatment with tibolone did not impact ApoA- II (n = 4 RCT arms, WMD: 1.32 mg/dL, 95 % CI: -4.39, 7.05, P = 0.64) and ApoB (n = 9 RCT arms, WMD: -2.68 mg/dL, 95 % CI: -20.98, 15.61, P = 0.77) values. In the subgroup analyses, we noticed a notable decrease in lipoprotein (a) levels when tibolone was prescribed to females aged < 60 years (WMD: -10.78 mg/dl) and when it was prescribed for ≤ 6 months (WMD: -15.69 mg/dl). The present meta-analysis of RCTs highlighted that treatment with tibolone reduces lipoprotein (a) and apolipoprotein A-I levels in postmenopausal women. As the decrease in serum lipids' concentrations is associated with a decrease in the risk of cardiovascular disease (CVD), treatment with tibolone could be a suitable therapy for postmenopausal women with elevated CVD risk.


Thoracic endografting increases cardiac afterload and leads to left ventricular hypertrophy in dogs.
Aortic endografting can cause aortic stiffening. We aimed to determine the chronic effect of thoracic endografting on cardiac afterload, function and remodelling. Eleven dogs were included, and all except 1 was successfully assessed [endograft, n = 5; sham operation (control), n = 5]. We deployed a stent graft in the descending aorta. The ascending aortic pressure and flow were measured, and aortic input impedance was obtained by frequency analysis to determine characteristic impedance and arterial compliance. Left ventricular pressure-volume relations were measured with an admittance catheter. Measurements were performed before, 10 min after and 3 months after endografting. Following euthanasia, we weighed the left ventricle of each dog and measured the cardiomyocyte cell size. Arterial compliance decreased from 0.47 ± 0.07 to 0.36 ± 0.06 and to 0.31 ± 0.05 ml/mmHg (both P < 0.01 versus baseline), and characteristic impedance increased from 0.11 ± 0.04 to 0.19 ± 0.05 and to 0.21 ± 0.04 mmHg/ml/s (both P < 0.01 versus baseline) 10 min and 3 months after endografting, respectively. Pressure-volume relation analysis showed that arterial elastance increased from 5.3 ± 1.0 to 6.7 ± 1.6 (at 10 min) and to 6.8 ± 1.0 mmHg/ml (at 3 months) (both P < 0.05 versus baseline), but end-systolic elastance and ventriculo-arterial coupling remained unchanged. Left ventricular weight to body weight ratio and left ventricular cardiomyocyte cell width in the endograft group were larger compared with the control's results (5.06 ± 0.27 g/kg vs 4.20 ± 0.49 g/kg, P = 0.009, 15.1 ± 1.7 µm vs 13.9 ± 1.5 µm, P = 0.02, respectively). The mid-term effect of the descending aortic endografting on left ventricular contractility and efficiency in canine normal hearts was minimal. However, endografting resulted in increased cardiac afterload and left ventricular hypertrophy.


Post-Loop Electrosurgical Excision Procedure High-Risk Human Papillomavirus Testing as a Test of Cure: The British Columbia Experience.
To determine whether Hybrid Capture 2 High-Risk HPV DNA Test (HC2) can be used as a test of cure in women treated for cervical intraepithelial neoplasia grade 2 or worse (CIN 2+) and allow discharge from colposcopy follow-up with a return to a cytology-based screening program for HC2-negative women. Data were analyzed for all women who underwent a loop electrosurgical excision procedure between August 1, 2008, and June 30, 2011, and had a valid HC2 result after loop electrosurgical excision procedure and follow-up histopathology result, to determine risk of persistent or recurrent CIN 2+ in HC2-positive and HC2-negative women. Two thousand three hundred forty women had adequate biopsies and valid HC2 results. Of 460 HC2-positive women, 118 (25.7%) were diagnosed with CIN 2+, whereas of 1,880 HC2-negative women, 35 (1.9%) had a subsequent diagnosis of CIN 2+ (p < .0002) yielding a HC2-negative predictive value of 98.1% (95% confidence interval = 97.4-98.7). Of 460 HC2-positive women, 306 initially had negative biopsies. In the subsequent 36 months, 38 of the 306 were diagnosed with CIN 2+. We conclude that women with a negative HC2 test can safely return to routine annual cytology screening by primary care providers while women who test HC2 positive are at higher risk and should continue to be followed by colposcopy, even if their initial biopsy is negative.


Impaired replication capacity of acute/early viruses in persons who become HIV controllers.
Human immunodeficiency virus type 1 (HIV-1) controllers maintain viremia at <2,000 RNA copies/ml without antiretroviral therapy. Viruses from controllers with chronic infection were shown to exhibit impaired replication capacities, in part associated with escape mutations from cytotoxic-T-lymphocyte (CTL) responses. In contrast, little is known about viruses during acute/early infection in individuals who subsequently become HIV controllers. Here, we examine the viral replication capacities, HLA types, and virus sequences from 18 HIV-1 controllers identified during primary infection. gag-protease chimeric viruses constructed using the earliest postinfection samples displayed significantly lower replication capacities than isolates from persons who failed to control viremia (P = 0.0003). Protective HLA class I alleles were not enriched in these early HIV controllers, but viral sequencing revealed a significantly higher prevalence of drug resistance mutations associated with impaired viral fitness in controllers than in noncontrollers (6/15 [40.0%] versus 10/80 [12.5%], P = 0.018). Moreover, of two HLA-B57-positive (B57(+)) controllers identified, both harbored, at the earliest time point tested, signature escape mutations within Gag that likewise impair viral replication capacity. Only five controllers did not express "protective" alleles or harbor viruses with drug resistance mutations; intriguingly, two of them displayed typical B57 signature mutations (T242N), suggesting the acquisition of attenuated viruses from B57(+) donors. These data indicate that acute/early stage viruses from persons who become controllers have evidence of reduced replication capacity during the initial stages of infection which is likely associated with transmitted or acquired CTL escape mutations or transmitted drug resistance mutations. These data suggest that viral dynamics during acute infection have a major impact on HIV disease outcome.


[Combined Effects of Soil Amendment and Zinc Fertilizer on Accumulation and Transportation of Cadmium in Soil-Rice System].
A field experiment was conducted in moderately and severely Cd contaminated paddy fields in Beishan Town, Changsha City, Hunan Province. This study examined the effects of LS amendment (limestone+sepiolite), in combination with soil application and foliar spraying of Zn fertilizer, on Cd uptake in early and late rice plants. The results showed that: ① the application of LS (2250 kg·hm-2 and 4500 kg·hm-2) significantly increased pH and CEC values in paddy soil during the early and late rice seasons, but the addition of Zn fertilizer (90 kg/hm2) to soil and through foliar spraying (0.2 g·L-1 and 0.4 g·L-1) had no significant effects on the pH or CEC of the soil. ② LS application decreased concentrations of TCLP-Cd and CaCl2-Cd in the soils, by 11.5%-38.8% and 24.0%-81.0%, respectively, while neither of the treatments involving the addition of Zn fertilizer to soil or through foliar spraying had any significant effects on the concentrations of TCLP-Cd and CaCl2-Cd. ③Single treatments involving only LS amendment, Zn fertilizer in soil, or foliar spraying of Zn fertilizer also reduced Cd concentrations in brown rice, but to a lesser degree than the combined treatments. The combined treatments (L1Z1F1, L1Z1F2, L2Z1F1, and L2Z1F2) reduced Cd concentrations in brown rice by 64.9%-67.5% and 56.1%-80.6%, for early and late rice, respectively, while L2Z1F1 (4500 kg·hm-2 LS+90 kg·hm-2 Zn fertilizer+foliar spraying 0.2 g·L-1 Zn fertilizer) resulted in the largest reduction in Cd concentration in brown rice. ④ The Cd/Zn ratio in brown rice was significantly positively correlated with Cd concentrations, indicating that increased Zn concentration in different rice tissues was one of the key reasons for decreased Cd concentration in brown rice. Clearly, as a remediation technology, combining LS amendments with zinc fertilizer is an effective method for achieving the safe utilization of moderately and severely Cd contaminated paddy fields, by effectively inhibiting the uptake, accumulation, and transportation of Cd in rice plants and decreasing Cd concentrations in brown rice.


THE IN VITRO DIFFERENTIATION OF MONONUCLEAR PHAGOCYTES. I. THE INFLUENCE OF INHIBITORS AND THE RESULTS OF AUTORADIOGRAPHY.
The influence of selected inhibitors of protein synthesis on the in vitro differentiation of mouse mononuclear phagocytes has been investigated. DL-p-Fluorophenylalanine at concentrations of 250 microg/ml inhibits the formation of three lysosomal hydrolases, cytochemically demonstrable acid phosphatase and osmiophilic, phase-dense granules. These effects occur in the absence of cell death and are reversed by L-phenylalanine. Puromycin at concentrations of 0.2 to 0.4 microg/ml has a similar effect on both the morphology and biochemistry of cell maturation. Colchicine at a concentration of 0.05 microg/ml inhibits the growth in cell diameter and has less of an effect on enzyme production. Cells exposed to leucine-H(3) for 2 to 3 minutes exhibit the localization of grains in the perinuclear-dense granule region. After an exposure of 60 minutes a similar localization is evident but with a correspondingly greater number of grains. A similar localization of grains occurs when choline-methyl-H(3) is employed as a tracer. The data suggest the storage of newly formed protein and possibly phospholipid in the centrosphere region.


The anti-inflammatory effects of acupuncture and their relevance to allergic rhinitis: a narrative review and proposed model.
Classical literature indicates that acupuncture has been used for millennia to treat numerous inflammatory conditions, including allergic rhinitis. Recent research has examined some of the mechanisms underpinning acupuncture's anti-inflammatory effects which include mediation by sympathetic and parasympathetic pathways. The hypothalamus-pituitary-adrenal (HPA) axis has been reported to mediate the antioedema effects of acupuncture, but not antihyperalgesic actions during inflammation. Other reported anti-inflammatory effects of acupuncture include an antihistamine action and downregulation of proinflammatory cytokines (such as TNF- α , IL-1 β , IL-6, and IL-10), proinflammatory neuropeptides (such as SP, CGRP, and VIP), and neurotrophins (such as NGF and BDNF) which can enhance and prolong inflammatory response. Acupuncture has been reported to suppress the expression of COX-1, COX-2, and iNOS during experimentally induced inflammation. Downregulation of the expression and sensitivity of the transient receptor potential vallinoid 1 (TRPV1) after acupuncture has been reported. In summary, acupuncture may exert anti-inflammatory effects through a complex neuro-endocrino-immunological network of actions. Many of these generic anti-inflammatory effects of acupuncture are of direct relevance to allergic rhinitis; however, more research is needed to elucidate specifically how immune mechanisms might be modulated by acupuncture in allergic rhinitis, and to this end a proposed model is offered to guide further research.


Isotype-specific immunoregulation; characterization and function of Fc receptors on T-T hybridomas which produce murine IgA-binding factor.
Several methods have been used in the present study to characterize Fc receptors (FcR) expressed on T-T hybridomas derived from mouse Peyer's patch T helper (Th) cell clones that preferentially support IgA responses. These T hybridomas (designated Th HA cells) produce IgA-binding factor (IBF alpha) which regulates antigen-dependent IgA responses. The ultrastructure of Th HA cells and the distribution of Fc alpha R on these cell lines were determined by colloidal gold (CG) immunoelectron microscopy (IEM). When Th HA cells were incubated with purified mouse IgA followed by CG-labeled anti-IgA, an even pattern of CG was distributed on the cell membrane. To ensure that binding occurred through Fc alpha R, Th HA cells were mixed with MOPC 315 IgA anti-DNP, followed by staining with CG-labeled TNP-human serum albumin. This resulted in an identical pattern of gold particle distribution, confirming expression of Fc alpha R on Th HA cells. No Fc mu R or Fc gamma 1R were detectable on Th HA cells by IEM. Immunocytoadherence with TNP-conjugated erythrocytes confirmed that Th HA cells were Fc alpha R+; however, no IgM or IgG rosettes were seen. When these cell lines were analyzed by flow cytometry (FACS) using IgA, IgM, or IgG1 and FITC-labeled anti-H chain-specific antibodies, 55 to 65% of cultured Th HA cells expressed Fc alpha R, and 11 to 18% expressed Fc mu R; however, no Fc gamma 1R was detectable on Th HA cells. The use of ELISA with Th HA cells as antigen confirmed the expression of Fc alpha R and the presence of less Fc mu R on these two cell lines. Solubilized membrane fractions derived from Th HA cells were tested for the presence of FcR by ELISA and for biologic function for support of IgA responses in Peyer's patch B cell cultures. Both Fc alpha R and Fc mu R were detected in fractions derived from Th HA cells. Furthermore, these fractions supported in vitro IgA anti-sheep erythrocyte responses, comparable to those obtained with Th HA cell culture supernatants containing IBF alpha. These studies show that Th HA cells express Fc alpha R with less Fc mu R, and the solubilized form of Fc alpha R exhibits IBF alpha-like activity. The significance of FcR expression by Th cell clones and cell lines and the relationship of soluble Fc alpha R and IBF alpha for IgA response regulation are discussed.


The adverse effect of benzhexol on memory in Parkinson's disease.
Seventy-eight subjects suffering from idiopathic Parkinson's disease were studied in 2 separate groups to test the hypothesis that benzhexol affects memory. In study A, 54 subjects were tested by means of a free recall technique; in study B, 24 subjects were given a signal detection memory task. Both studies yielded significant correlations between memory impairment and dosage levels of benzhexol. No correlation was found between memory status and dosage of levodopa, duration of illness or Hamilton depression scores. The implications of these findings are discussed.


A longitudinal assessment of antimicrobial susceptibility among important pathogens collected as part of the Tigecycline Evaluation and Surveillance Trial (T.E.S.T.) in France between 2004 and 2012.
Clinically important Gram-positive and -negative isolates were collected from patients in France between 2004 and 2012 as a part of the Tigecycline Evaluation and Surveillance Trial. MICs were determined using methodology described by the Clinical and Laboratory Standards Institute. In total, 17,135 isolates were contributed by 29 medical centres; respiratory (25.1%) and cardiovascular (20.3%) sources predominated. High susceptibility was observed among Enterococcus spp. and Staphylococcus aureus (including methicillin-resistant S. aureus [MRSA]) to linezolid (100%), tigecycline (≥99.8%) and vancomycin (≥94.6%). The percentage of MRSA decreased from 34.3% in 2004 to 20.0% in 2009 before increasing to 34.7% in 2012. Vancomycin, linezolid, levofloxacin and carbapenems were highly active (≥99.6%) against Streptococcus pneumoniae; 3.2% were PRSP. Escherichia coli showed peak susceptibility to the carbapenems (≥99.9%), tigecycline (99.3%) and amikacin (97.9%); significant (p < 0.01) decreases in susceptibility were observed for ampicillin, cefepime and ceftriaxone between 2004 and 2012. ESBL production among E. coli increased from 3.0% (2004) to 14.9% (2012). High susceptibility was noted among Haemophilus influenzae to levofloxacin (100%), amoxicillin-clavulanate (99.2%), carbapenems (≥98.7%) and ceftriaxone (98.5%); β-lactamase production fluctuated with no notable trend between 18.1% (2007) and 27.7% (2011). Klebsiella spp. were highly susceptible to carbapenems (≥99.6%) and amikacin (≥96.4%); significant (p < 0.01) decreases in amoxicillin-clavulanate, cefepime, ceftriaxone, levofloxacin, piperacillin-tazobactam and tigecycline susceptibility were observed among K. pneumoniae between 2004 and 2012. Only imipenem was highly active (96.5% susceptible) against Acinetobacter baumannii. Imipenem and amikacin (87.7% and 87.1% susceptible) were the most active agents against P. aeruginosa; 10.2% of isolates were categorized as multidrug resistant. Carbapenems, linezolid, tigecycline and vancomycin conserved good in vitro activity against most pathogens (according to their spectrum of activity) in France between 2004 and 2012.


Single Tooth Prosthetic Restoration Through Surgical Crown Lengthening, Conservative Therapies and CAD-CAM Milled Restoration in Lithium-Disilicate. A Case Report.
The restorative treatment of a decayed, non-vital upper premolar often requires an interdisciplinary approach. Esthetics and the entity of the masticatory loads are usually affecting the choice of procedures and materials. A conservative treatment in a case with esthetic needs and functional requirements is investigated in this case report. A case of a 45- year- old woman with a severely decayed upper premolar is presented. The tooth has been previously treated in an emergency department with a provisional endodontic medication, but a further treatment has been then performed. The tooth was painful at the moment of the visit and the carious lesion appeared to invade the subgingival level. A surgical-conservative approach has been selected involving crown lengthening, endodontic treatment and a prosthetic crown with a monolithic Lithium-Silicate. The rehabilitative process required 3 months, including maturation times after surgery and the placement of the provisional crown. The patient claimed to be both esthetically and functionally satisfied with the restoration. A conservative treatment of a severely involved tooth requires a critical evaluation of the remaining structures and a precise selection of the restorative materials. A monolithic crown could represent an ideal solution for restoring an upper premolar requiring esthetics and solid function, but the choice of a high translucent, easily polishable and possibly even chair-side prosthetic material might be an appreciable added value for clinicians.


Loperamide-Induced Torsades de Pointes: A Case Series.
Loperamide is an over-the-counter, inexpensive, antidiarrheal opioid that can produce life-threatening toxicity at high concentrations. CASE REPORT 1: A 28-year-old man with a history of depression and substance abuse disorder (SUD) presented to the emergency department (ED) with shortness of breath and lightheadedness. He ingested large amounts of loperamide daily. The patient's initial electrocardiogram (ECG) demonstrated sinus rhythm, right axis deviation, undetectable PR interval, QRS 168 ms, and QTc 693 ms. He was administered intravenous sodium bicarbonate and magnesium sulfate and admitted to the intensive care unit, eventually developing Torsades de Pointes (TdP). He was given lidocaine and isoproterenol infusions, and an external pacemaker was placed. He was discharged in stable condition on hospital day (HD) 16. CASE REPORT 2: A 39-year-old woman with a history of hepatitis C, depression, and SUD was transported to the ED after reported seizure-like activity. The patient experienced TdP in the ED and admitted to ingesting large amount of loperamide daily. An ECG demonstrated sinus rhythm, right axis deviation, PR interval 208 ms, QRS interval 142 ms, and QTc 687 ms. She was administered intravenous magnesium, sodium bicarbonate, and isoproterenol. After intensive care unit admission, the patient experienced no further TdP and was discharged on HD 6. WHY SHOULD AN EMERGENCY PHYSICIAN BE AWARE OF THIS?: Emergency physicians should proceed with caution when treating patients with loperamide toxicity. Even in asymptomatic patients and drug discontinuance, obtain consultation with a medical toxicologist, promptly treat ECG abnormalities aggressively, and admit all patients for further monitoring.


Characterization of a new GlnR binding box in the promoter of amtB in Streptomyces coelicolor inferred a PhoP/GlnR competitive binding mechanism for transcriptional regulation of amtB.
The transcription of amtB in Streptomyces coelicolor has been proposed to be counter-regulated by GlnR (a global regulator for nitrogen metabolism) and PhoP (a global regulator for phosphate metabolism). However, the GlnR-protected region, which was deduced to be two 22-bp GlnR binding boxes (gTnAc-n6-GaAAc-n6-GtnAC-n6-GAAAc-n6, abbreviated as a1-b1 and a2-b2), was separated from the PhoP-protected region in the promoter of amtB, leaving the mechanism for this regulation undefined. In this study, another 22-bp GlnR binding box, which consisted of a3-site-n6-b3-site (a3-b3) overlapping with the PhoP-binding sequences, was identified in the promoter region of amtB by a DNase I footprinting assay. An electrophoretic mobility shift assay (EMSA) using purified recombinant GlnR and the synthetic amtB promoter fragments with the three GlnR binding boxes individually mutated demonstrated that every box was involved in GlnR binding in vitro. Further in vivo assays using the egfp reporter gene fused to various kinds of mutated promoter regions of amtB demonstrated that all of the three GlnR binding boxes were required for GlnR-mediated activation of amtB transcription under the nitrogen-limited condition. The results of EMSA using the amtB promoter with mixtures of recombinant His-tagged GlnR and Trx-His-S-tagged PhoP inferred that PhoP might compete against GlnR from binding at the a3-b3 site, attributable to the PhoP/GlnR counter-regulatory function subjected to further experimental proof.


Transendothelial chemotaxis of human alpha/beta and gamma/delta T lymphocytes to chemokines.
Two subpopulations of human T lymphocytes expressing different antigen receptors, alpha/beta and gamma/delta, emigrate into inflamed tissues in distinctive patterns. We compared the transmigration of alpha/beta and gamma/delta T cells to C-C and C-X-C chemokines using an in vitro transendothelial chemotaxis assay. The C-C chemokines monocyte chemoattractant protein (MCP)-1, RANTES, macrophage inflammatory protein (MIP)-1alpha and MIP-1beta stimulated similar, dose-dependent chemotaxis of purified gamma/delta T cells, whereas MCP-1, RANTES, and MIP-1alpha produced greater chemotaxis of purified alpha/beta T cells than MIP-1beta. In contrast, the C-X-C chemokines interleukin (IL)-8 and interferon-gamma inducible protein-10 (IP-10) did not promote chemotaxis of either alpha/beta or gamma/delta T cells. Three gamma/delta T cell clones with differing CD4 and CD8 phenotypes also migrated exclusively to C-C chemokines. Phenotypic analysis of mononuclear cells that transmigrated from an input population of unfractionated peripheral blood mononuclear cells confirmed the results with purified gamma/delta T cells. Our data demonstrate that human peripheral blood alpha/beta and gamma/delta T cells can transmigrate to MCP-1, RANTES, MIP-1alpha, and MIP-1beta, and suggest that both T lymphocyte subpopulations share the capacity to emigrate in response to C-C chemokines during inflammation.


Prediction of Aqueous Reduction Potentials of X•, ChH•, and XO• Radicals with X = Halogen and Ch = Chalcogen.
The aqueous electron affinity and aqueous reduction potentials for F•, Cl•, Br•, I•, OH•, SH•, SeH•, TeH•, ClO•, BrO•, and IO• were calculated using electronic structure methods for explicit cluster models coupled with a self-consistent reaction field (SMD) to treat the aqueous solvent. Calculations were conducted using MP2 and correlated molecular orbital theory up to the CCSD(T)-F12b level for water tetramer clusters and MP2 for octamer cluster. Inclusion of explicit waters was found to be important for accurately predicting the redox potentials in a number of cases. The calculated reduction potentials for X• and ChH• were predicted to within ∼0.1 V of the reported literature values. Fluorine is anomalous due to abstraction of a hydrogen from one of the surrounding water molecules to form a hydroxyl radical and hydrogen fluoride, so its redox potential was calculated using only an implicit model. Larger deviations from experiment were predicted for ClO• and BrO•. These deviations are due to the free energy of solvation of the anion being too negative, as found in the pKa calculations, and that for the neutral being too positive with the current approach.


Severe acute respiratory syndrome caused by the influenza A (H1N1) virus.
In view of the pandemic caused by a new virus, influenza A (H1N1), we report the case of a 56-year-old patient without relevant risk factors and with severe acute respiratory syndrome resulting from infection with this virus. We present the results of laboratory tests and the imaging findings (chest X-ray and CT scans). The evolution was favorable, and the patient was discharged after 14 days.


[Infectious complications in cesarean section: previous classification and use of antibiotics].
A retrospective study of all the patients that underwent cesarean section at "Ramón Gonzalez Coro" Gynecoobstetric Hospital in 1997 and 1998 was made. The information about some variables, such as the diagnosis of cesarean sections, the frequency of infectious complications, the presence of complications according to the classification, the correct and incorrect use of antibiotics according to the classification of the surgical procedure and the occurrence of infectious complications due to the correct or incorrect use of antibiotics that were collected in the database of the Automated System to register cesarean sections was processed. The main indication was anterior cesarean section (34.1%), followed by cephalopelvic disproportion (13.5%). 17.1% had some infectious complications. Endometritis was the most frequent complicatrion (7.9%). Most of the infectious complications occurred in the clean cesarean sections (16.7%) and in the dirty cesarean sections (22.3%). The best use of antibiotics was observed in both groups (96.5 and 96.6%, respectively). Patients with an incorrect use of antibiotics had more infectious complications (37.8%). A statistical association was found between these 2 variables (p = 7.569 x 20).


Superficial femoral artery as inflow for bypass to the proximal popliteal artery.
We have obtained long term follow-up on 58 patients in whom the superficial femoral artery was used as a donor site for bypass to the proximal popliteal artery. The indication for the reconstructive procedure was intermittent claudication 36%, rest pain 34%, and gangrene 26%. All patients had angiographic evidence of a patent proximal superficial femoral artery and many had relative indications for short bypass such as limited saphenous vein availability or compromised medical condition. There was a cumulative patency rate of 79% by life table method for the series with a six year follow-up. We have observed that the superficial femoral artery is a satisfactory inflow site for proximal popliteal bypass grafts in selected patients. The procedure can maximize use of a limited segment of vein with minimal operative time and morbidity. Inflow stenosis can be revised or long bypass procedures can be offered in the event of graft failure.


VarQ: A Tool for the Structural and Functional Analysis of Human Protein Variants.
Understanding the functional effect of Single Amino acid Substitutions (SAS), derived from the occurrence of single nucleotide variants (SNVs), and their relation to disease development is a major issue in clinical genomics. Despite the existence of several bioinformatic algorithms and servers that predict if a SAS is pathogenic or not, they give little or no information at all on the reasons for pathogenicity prediction and on the actual predicted effect of the SAS on the protein function. Moreover, few actual methods take into account structural information when available for automated analysis. Moreover, many of these algorithms are able to predict an effect that no necessarily translates directly into pathogenicity. VarQ is a bioinformatic pipeline that incorporates structural information for the detailed analysis and prediction of SAS effect on protein function. It is an online tool which uses UniProt id and automatically analyzes known and user provided SAS for their effect on protein activity, folding, aggregation and protein interactions, among others. We show that structural information, when available, can improve the SAS pathogenicity diagnosis and more important explain its causes. We show that VarQ is able to correctly reproduce previous analysis of RASopathies related mutations, saving extensive and time consuming manual curation. VarQ assessment was performed over a set of previously manually curated RASopathies (diseases that affects the RAS/MAPK signaling pathway) related variants, showing its ability to correctly predict the phenotypic outcome and its underlying cause. This resource is available online at http://varq.qb.fcen.uba.ar/. Supporting Information & Tutorials may be found in the webpage of the tool.


Semliki forest virus vectors engineered to express higher IL-12 levels induce efficient elimination of murine colon adenocarcinomas.
To evaluate the use of alphavirus vectors for tumor treatment we have constructed and compared two Semliki Forest virus (SFV) vectors expressing different levels of IL-12. SFV-IL-12 expresses both IL-12 subunits from a single subgenomic promoter, while in SFV-enhIL-12 each IL-12 subunit is expressed from an independent subgenomic promoter fused to the SFV capsid translation enhancer. This latter strategy provided an eightfold increase of IL-12 expression. We chose the poorly immunogenic MC38 colon adenocarcinoma model to evaluate the therapeutic potential of SFV vectors. A single intratumoral injection of 10(8) viral particles of SFV-IL-12 or SFV-enh-IL-12 induced>or=80% complete tumor regressions with long-term tumor-free survival. However, lower doses of SFV-enhIL-12 were more efficient than SFV-IL-12 in inducing antitumoral responses, indicating a positive correlation between the IL-12 expression level and the therapeutic effect. Moreover, repeated intratumoral injections of suboptimal doses of SFV-enhIL-12 increased the antitumoral response. In all cases SFV vectors were more efficient at eliminating tumors than a first-generation adenovirus vector expressing IL-12. In addition, the antitumoral effect of SFV vectors was only moderately affected by preimmunization of animals with high doses of SFV vectors. This antitumoral effect was produced, at least partially, by a potent CTL-mediated immune response.


Frequency of head-impact-related outcomes by position in NCAA division I collegiate football players.
Concussions and subconcussive impacts sustained in American football have been associated with short- and long-term neurological impairment, but differences in head impact outcomes across playing positions are not well understood. The American Medical Society for Sports Medicine has identified playing position as a key risk factor for concussion in football and one for which additional research is needed. This study examined variation in head impact outcomes across primary football playing positions in a group of 730 National Collegiate Athletic Association Division I Football Championship Series athletes, using a self-report questionnaire. Although there were no significant differences between position groups in the number of diagnosed concussions during the 2012 football season, there were significant differences between groups in undiagnosed concussions (p=0.008) and "dings" (p<0.001); offensive linemen reported significantly higher numbers than most other positions. Significant differences were found between position groups in the frequencies of several postimpact symptoms, including dizziness (p<0.001), headache (p<0.001), and seeing stars (p<0.001) during the 2012 football season, with offensive linemen reporting significantly more symptoms compared to most other groups. There were also positional differences in frequency of returning to play while symptomatic (p<0.001) and frequency of participating in full-contact practice (p<0.001). Offensive linemen reported having returned to play while experiencing symptoms more frequently and participating in more full-contact practices than other groups. These findings suggest that offensive linemen, a position group that experiences frequent, but low-magnitude, head impacts, develop more postimpact symptoms than other playing positions, but do not report these symptoms as a concussion.


Environmental performance and mechanical analysis of concrete containing recycled asphalt pavement (RAP) and waste precast concrete as aggregate.
The overall objective of this research project was to investigate the feasibility of incorporating 100% recycled aggregates, either waste precast concrete or waste asphalt planning, as replacements for virgin aggregates in structural concrete and to determine the mechanical and environmental performance of concrete containing these aggregates. Four different types of concrete mixtures were designed with the same total water cement ratio (w/c=0.74) either by using natural aggregate as reference or by totally replacing the natural aggregate with recycled material. Ground granulated blast furnace slag (GGBS) was used as a mineral addition (35%) in all mixtures. The test results showed that it is possible to obtain satisfactory performance for strength characteristics of concrete containing recycled aggregates, if these aggregates are sourced from old precast concrete. However, from the perspective of the mechanical properties, the test results indicated that concrete with RAP aggregate cannot be used for structural applications. In terms of leaching, the results also showed that the environmental behaviour of the recycled aggregate concrete is similar to that of the natural aggregate concrete.


[cGMP-activated phosphodiesterase from human brain: kinetic and regulatory properties].
The kinetic and regulatory properties of cGMP-activated phosphodiesterase (PDE) from human brain were studied. In double reciprocal plots the enzyme activity is characterized by a linear dependence of cAMP and a nonlinear one for cGMP. Micromolar concentrations of cGMP accelerate cAMP hydrolysis (7-14-fold) with Ka for cGMP of 0.36 microM. Stimulation of cAMP hydrolysis is accompanied by a decrease of Km with no changes in Vmax. With a rise in the cGMP concentration above 5 microM PDE activation is changed by its inhibition. Both substrates act as competitive inhibitors towards each other. The Ki value for both cGMP and cAMP is 30 microM. After the increase in the cAMP (Bt)2 concentration the activation of 5 microM cAMP hydrolysis is accompanied by the enzyme inhibition. Both analogs competitively inhibit cGMP hydrolysis with Ki of 10 and 1500 microM for cGMP(Bt)2 and cAMP(Bt)2, respectively. The data obtained point to the existence of two binding sites for cyclic nucleotides, namely, a regulatory site which is highly specific for cGMP and a catalytic site responsible for the hydrolysis of the both substrates which displays no apparent specificity either for cAMP or for cGMP. The different affinity of natural and synthetic cyclic nucleotides for these sites is determined, to a large extent, by the amino groups in the 2nd and 6th positions of the purine ring.


The Health Buddies App as a Novel Tool to Improve Adherence and Knowledge in Atrial Fibrillation Patients: A Pilot Study.
Atrial fibrillation (AF) constitutes an important risk for stroke, especially in an ageing population. A new app (Health Buddies) was developed as a tool to improve adherence to non-vitamin K antagonist oral anticoagulants (NOACs) in an elderly AF population by providing a virtual contract with their grandchildren, spelling out daily challenges for both. The aim of this pilot study was to assess the feasibility and usability of the Health Buddies app in AF patients. Two workshops were conducted to steer app development and to test a first prototype. The feasibility of the finalized app was investigated by assessing the number of eligible AF patients (based on current prescription of NOACs, the presence of grandchildren between 5 and 15 years old, availability of a mobile phone, computer, or tablet), and the proportion of those who were willing to participate. Participants had to use the app for 3 months. The motivation of the patients to use the app was assessed based on the number of logins to the app. Their perception of its usefulness was examined by specific questionnaires. Additionally, the effects on knowledge level about AF and its treatment, and adherence to NOAC intake were investigated. Out of 830 screened AF patients, 410 were taking NOACs and 114 were eligible for inclusion. However, only 3.7% (15/410) of the total NOAC population or 13.2% of the eligible patients (15/114) were willing to participate. The main reasons for not participating were no interest to participate in general or in the concept in particular (29/99, 29%), not feeling comfortable using technology (22/99, 22%), no interest by the grandchildren or their parents (20/99, 20%), or too busy a lifestyle (12/99, 12%). App use significantly decreased towards the end of the study period in both patients (P=.009) and grandchildren (P<.001). NOAC adherence showed a taking adherence and regimen adherence of 88.6% (SD 15.4) and 81.8% (SD 18.7), respectively. Knowledge level increased from 64.6% (SD 14.7) to 70.4% (SD 10.4) after 3 months (P=.09). The app scored positively on clarity, novelty, stimulation, and attractiveness as measured with the user experience questionnaire. Patients evaluated the educational aspect of this app as a capital gain. Only a small proportion of the current AF population seems eligible for the innovative Health Buddies app in its current form. Although the app was positively rated by its users, a large subset of patients was not willing to participate in this study or to use the app. Efforts have to be made to expand the target group in the future.


Empirical LiK excited state potentials: connecting short range and near dissociation expansions.
We report on a high-resolution spectroscopic survey of 6Li40K molecules near the 2S + 4P dissociation threshold and produce a fully empirical representation for the B1Π potential by connecting available short- and long-range data. The purpose is to identify a suitable intermediate state for a coherent Raman transfer to the absolute ground state, and the creation of a molecular gas with dipolar interactions. Starting from weakly bound ultracold Feshbach molecules, the transition frequencies to twenty-six vibrational states are determined. Our data are combined with long-range measurements [Ridinger et al., EPL, 2011, 96, 33001], and near-dissociation expansions for the spin-orbit coupled potentials are fitted to extract the van der Waals C6 dispersion coefficients. A suitable vibrational level is identified by resolving its Zeeman structure and by comparing the experimentally attained g-factor to our theoretical prediction. Using mass-scaling of the short-range data for the B1Π [Pashov et al., Chem. Phys. Lett., 1998, 292, 615620] and an updated value for its depth, we model the short- and the long-range data simultaneously and produce a Rydberg-Klein-Rees curve covering the entire range.


Comparative study on antigenicity and immunogenicity of 26-28 kDa antigen and recombinant Sj26 (RSj26) of Schistosoma japonicum.
This paper reports a comparison of the recombinant Sj26 (rSj26) antigen derived from the Philippine strain and the 26-28 kDa antigen isolated and purified from the Chinese strain of Schistosoma japonicum with respect to their antigenicity and immunogenicity. The results showed that there were obvious cross reactions between rSj26 and 26-28 kDa antigen when rSj26 antigen was tested against specific antibodies in sera of mice infected with the Chinese strain of S. japonicum or the 26-28 kDa antigen was tested against specific anti-rSj26 antibodies by ELISA, IFA and Western blotting. Both the 26-28 kDa and the rSj26 antigen had weak cross reactions with SEA antigen. The worm reduction rate after challenging with Chinese strain cercariae in mice immunized with rSj26 was 26-32%, similar to that in mice immunized with 26-28 kDa antigen. It is suggested that rSj26 antigen can induce a certain level of specific protective immunity in the host against infection by the Chinese strain of S. japonicum cercariae.


5-HT1F receptor regulates mitochondrial homeostasis and its loss potentiates acute kidney injury and impairs renal recovery.
Our laboratory previously reported that agonists of the 5-hydoxytryptamine 1F (5-HT1F) receptor induce renal mitochondrial biogenesis (MB) and that stimulation of the 5-HT1F receptor following ischemia/reperfusion (I/R)-induced acute kidney injury (AKI) accelerated the recovery of renal function in mice. The goal of this study was to examine the contribution of the 5-HT1F receptor in the regulation of renal mitochondrial homeostasis and renal function in naïve and injured mice. Although 5-HT1F receptor knockout (KO) mice were healthy and fertile, and did not exhibit renal dysfunction, renal mitochondrial DNA copy number and mitochondrial fission gene expression increased at 10 wk of age. The 5-HT1F receptor KO mice exhibited greater proximal tubular injury and diminished renal recovery after I/R-induced AKI compared with wild-type mice. These findings were associated with persistent suppression of renal cortical MB and ATP levels after injury. In summary, the 5-HT1F receptor is a component of physiological MB regulation in the kidney, and its absence potentiates renal injury and impedes recovery.


To be precise, the details don't matter: On predictive processing, precision, and level of detail of predictions.
Many theoretical and empirical contributions to the Predictive Processing account emphasize the important role of precision modulation of prediction errors. Recently it has been proposed that the causal models used in human predictive processing are best formally modeled by categorical probability distributions. Crucially, such distributions assume a well-defined, discrete state space. In this paper we explore the consequences of this formalization. In particular we argue that the level of detail of generative models and predictions modulates prediction error. We show that both increasing the level of detail of the generative models and decreasing the level of detail of the predictions can be suitable mechanisms for lowering prediction errors. Both increase precision, yet come at the price of lowering the amount of information that can be gained by correct predictions. Our theoretical result establishes a key open empirical question to address: How does the brain optimize the trade-off between high precision and information gain when making its predictions?


Balanced Reciprocal Translocation t(X;1) in a Girl with Tall Stature and Primary Amenorrhea.
Chromosomal translocations constitute one of the most important, yet uncommon, causes of primary amenorrhea and gonadal dysgenesis. Although X-autosome translocations are frequently associated with streak gonads and clinical features of the Turner syndrome, the majority of X-autosome carriers may present with a variable phenotype, developmental delay, and recognizable X-linked syndrome due to nonrandom X-inactivation. In this article, we describe a healthy 15.5-year-old girl with primary amenorrhea, gonadal dysgenesis, and tall stature without other manifestations of the Turner syndrome. Relevant clinical, biochemical, endocrinological, and cytogenetical evaluations were performed. Initial investigations revealed hypergonadotropic hypogonadism (FSH=134 mIU/mL [normal=10-15 mIU/mL], LH=47.5 [normal=10-15 mIU/mL], and estradiol=24.3 pmol/L). On ultrasound examination of the pelvis, streak ovaries with a hypoplastic uterus were noted. Chromosome study, performed according to routine procedures, revealed an apparently balanced reciprocal translocation involving the short arm of chromosome 1(p2) and the long arm of the X chromosome (q2) in all the cells with the following karyotype: 46,X,t(1;X)(p13;q22). She was placed on hormone replacement therapy. In our patient, X-autosome translocation was associated with gonadal dysgenesis and tall stature. We conclude that t(X;1) may be associated with gonadal dysgenesis without other congenital abnormalities. To our knowledge, normal phenotype with gonadal dysgenesis and tall stature in association with t(X;1) translocation has not been previously reported.


Balloon atrial septostomy guided by echocardiography in a neonatal intensive care unit.
The bedside two-dimensional echocardiography (2-D ECHO) has been successfully used to guide the balloon atrial septostomy, speeding up the procedure and preventing the risks of transportation to the hemodynamics laboratory. To assess the results of the bedside balloon atrial septostomy in cyanotic neonates. Between January/1997 and July/2008, 102 atrioseptostomies by balloon catheter guided by echocardiography were carried out and saturation levels, defect diameter, oxygen saturation, clinical-laboratory response and complications related to the procedure were evaluated. Of 102 cases of bedside Rashkind septostomy, 98 met the inclusion criteria, with 90 neonates in Group A (Rashkind procedure in the preoperative phase) and 8 in Group B (procedure at the postoperative phase). There was a predominance of the male sex (75%), mean age was 8.3 ± 9.3 days and the mean weight was 3,100 ± 1,100 g; the transposition of the great arteries was the most frequent congenital heart disease (n = 74). When comparing the levels of saturation pre and post-procedure (65.9 ± 19.5% and 86 ± 9.7%) and the diameter of the interatrial septal defect pre and post-procedure (2.3 ± 1.0 mm and 5.5 ± 1.3 mm) there was a statistically significant difference (p < 0.001). When comparing the levels of saturation and the diameter of the interatrial septal defect between the group of survivors vs non-survivor, there was no statistically significant difference (p > 0.05). The balloon atrial septostomy guided by the echocardiography reduces the risks by allowing the effective palliative procedure to be carried out promptly in cyanotic neonates, in addition to presenting lower hospital costs.


Safety and Pharmacokinetics of DS-1040 Drug-Drug Interactions With Aspirin, Clopidogrel, and Enoxaparin.
DS-1040, a novel low-molecular-weight inhibitor of activated thrombin-activatable fibrinolysis inhibitor, is under development for the treatment of thromboembolic diseases including venous thromboembolism and acute ischemic stroke. Here we describe the results of 3 studies that evaluated the safety and tolerability of DS-1040 along with the effect on DS-1040 pharmacokinetic (PK) parameters, when dosed alone or when coadministered with aspirin (NCT02071004), clopidogrel (NCT02560688), or enoxaparin in healthy subjects. Concomitant administration of single-dose DS-1040 with multiple-dose aspirin, multiple-dose clopidogrel, or single-dose enoxaparin, consistent with clinically relevant dose regimens, was safe and well tolerated with no serious treatment-emergent adverse events (TEAEs), TEAEs leading to discontinuation, bleeding-related TEAEs, and no significant changes in coagulation parameters. DS-1040 did not prolong bleeding time when administered concomitantly with aspirin or clopidogrel. In the aspirin study, DS-1040 PK was evaluated following the concomitant administration with multiple-dose aspirin, where the plasma DS-1040 exposure (peak plasma concentration [Cmax ] and area under the concentration-time curve [AUCinf ]) was to be similar to the data previously published in the first-in-human study of DS-1040 in healthy subjects. The PK parameters of DS-1040 coadministered with clopidogrel were similar to those of DS-1040 alone, with small increases in geometric means for Cmax (7%) and AUClast (9%). When coadministered with enoxaparin, the PK parameters of DS-1040 were not affected (1.1% and 1.5% decreases in geometric means for Cmax and AUClast , respectively). Therefore, concomitant administration of DS-1040 and clopidogrel or enoxaparin did not demonstrate PK drug-drug interactions.


Nutritional targeting of cyclooxygenase-2 for colon cancer prevention.
Factors related to diet and life style have been identified as primary determinants in about 80% of colorectal cancers. Non-steroidal anti-inflammatory drugs (NSAID) and selective cyclooxygenase-2 (COX-2) inhibitors (COXIB) reduce the relative risk of colon cancer. To overcome systemic COX inhibition associated with NSAID and COXIB, there is a growing interest in developing alternative colon cancer prevention strategies using diet-based approaches that target COX-2. The transition from aberrant crypt foci (ACF) to colon cancer is a multiyear process providing opportunities for nutritional targeting of genes influencing the course of this disease process at early stages of development. The activation of the proinflammatory gene COX-2 and PG production in the colonic mucosa are recognized risk factors in colon cancer. Many natural food components may impact colon cancer risk by interfering with ligand-activated receptors, signal transduction pathways, and transcription factors involved in stimulation of COX-2 expression. In this review, we highlight key upstream features of signaling pathways and transcriptional control of the COX-2 gene and discuss opportunities for dietary modulation of COX-2 expression in gastro-intestinal cancers with special emphasis on prevention of colorectal tumors. Review of the experimental evidence suggests that dietary strategies based on specific or cocktails of bioactive food components as well nutritional-pharmacological combinations targeted to regulation of COX-2 expression and activity may prove useful in the prevention of colon cancer. An integrated approach may offer the advantage of combined higher efficacies. Future studies should investigate the efficacy of combinations of bioactive food compounds on epigenetic regulation of the COX-2 gene and characterize potential synergies and amplification effects resulting from the concomitant use of bioactive food components and COX-2 inhibitors.


Multivariate analysis of the volatile components in tobacco based on infrared-assisted extraction coupled to headspace solid-phase microextraction and gas chromatography-mass spectrometry.
A novel infrared-assisted extraction coupled to headspace solid-phase microextraction followed by gas chromatography with mass spectrometry method has been developed for the rapid determination of the volatile components in tobacco. The optimal extraction conditions for maximizing the extraction efficiency were as follows: 65 μm polydimethylsiloxane-divinylbenzene fiber, extraction time of 20 min, infrared power of 175 W, and distance between the infrared lamp and the headspace vial of 2 cm. Under the optimum conditions, 50 components were found to exist in all ten tobacco samples from different geographical origins. Compared with conventional water-bath heating and nonheating extraction methods, the extraction efficiency of infrared-assisted extraction was greatly improved. Furthermore, multivariate analysis including principal component analysis, hierarchical cluster analysis, and similarity analysis were performed to evaluate the chemical information of these samples and divided them into three classifications, including rich, moderate, and fresh flavors. The above-mentioned classification results were consistent with the sensory evaluation, which was pivotal and meaningful for tobacco discrimination. As a simple, fast, cost-effective, and highly efficient method, the infrared-assisted extraction coupled to headspace solid-phase microextraction technique is powerful and promising for distinguishing the geographical origins of the tobacco samples coupled to suitable chemometrics.


Failure of guanidine and 2-(alpha-hydroxybenzyl)benzimidazole to inhibit replication of hepatitis A virus in vitro.
Replication of hepatitis A virus (HAV) in the human hepatoma-derived PLC/PRF/5 cell line was neither inhibited in the presence of various concentrations of guanidine or D-2-(alpha-hydroxybenzyl)benzimidazole (D-HBB), nor were the two chemicals effective in combination. Under identical conditions, however, replication of poliovirus type 1 was inhibited. Tracer experiments with radiolabelled guanidine and D-HBB also furnished no evidence that the two antiviral substances were metabolized gradually to inactive derivatives in PLC/PRF/5 cells. Therefore, it is concluded that resistance to the action of guanidine and D-HBB is an inherent characteristic of HAV. However, the insensitivity of HAV to these drugs does not exclude the virus from the family of picornaviruses.


Beta-adrenergic blocking agents in the treatment of patients after a myocardial infarction.
Beta-adrenergic blocking agents have been widely used in ischemic heart disease. They have achieved their greatest benefit in the secondary prevention of recurrent events in patients following acute myocardial infarction (MI). This is a review of the major clinical investigations exploring the effects of beta-adrenergic blocking agents in patients following acute MI and in a variety of patient subsets. These data indicate that the routine use of beta-adrenergic blocking agents in postinfarction patients results in a 25% to 35% decrease in mortality and has increased relative and absolute benefit in patients with ventricular ectopy and left ventricular dysfunction. The adverse effects of beta-adrenergic blocking agents are discussed which indicate that these drugs are well tolerated with little or no side effects. This review supports the observation that beta-adrenergic blocking agents have an important role in the treatment of patients following an acute MI, with the exclusion of those with chronic lung disease and severe left ventricular dysfunction.


proANP Metabolism Provides New Insights Into Sacubitril/Valsartan Mode of Action.
Sacubitril/valsartan (S/V) treatment is beneficial in patients with heart failure with reduced ejection fraction (HFrEF), but its mode of action remains elusive, although it involves the increase in ANP (atrial natriuretic peptide). Combining mass spectrometry and enzymatic assay in the plasma of 73 HFrEF patients treated with S/V and controls, we deciphered proANP processing that converts proANP into 4 vasoactive peptides. We found that proANP processing is sequential and involved meprin B, ECE (endothelin-converting enzyme) 1, and ANPEP (aminopeptidase N). This processing is limited in HFrEF patients via the downregulation of proANP production, corin, and meprin B activities by miR-425 and miR1-3p. S/V restored or compensated proANP processing by downregulating miR-425 and miR1-3p, hence increasing levels of proANP-derived bioactive peptides. In contrast, S/V directly and indirectly partially inhibited ECE1 and ANPEP. ECE1 partial inhibition resulted in a lower-than-expected increase in ET1 (endothelin 1), tilting the vasoactive balance toward vasodilation, and possibly hypotension. Furthermore, proANP glycosylation interferes with the midregional proANP assay -a clinical surrogate for proANP production, preventing any pathophysiological interpretation of the results. The analysis of S/V dose escalation with respect to baseline treatments suggests S/V-specific effects. These findings offer mechanistic evidence to the natriuretic peptide -defective state in HFrEF, which is improved by S/V. These data also strongly suggests that S/V increases plasma ANP by multiple mechanisms that involve 2 microRNAs, besides its protection from NEP (neprilysin) cleavage. Altogether, these data provide new insights on HFrEF pathophysiology and the mode of action of S/V.


Paralytic shellfish poisoning: a review.
Paralytic shellfish poisoning (PSP) in man results from the consumption of mussels, clams, and oysters that have fed on toxic dinoflagellates. Motile, marine protozoa of the dinoflagellate group often produce "blooms," i.e., red tides, which color the sea. Not all genera or species are toxic to fish and mammals, nor are the toxic principles the same in all poisonous protozoa. At least 5 of the group are known to cause poisonings in man. Shellfish poisonings other than PSP are also recognized. The PSP toxin, saxitoxin, is concentrated in the viscera and occasionally in the mantle and syphon of marine bivalves. Cooking does not completely destroy the low molecular weight poisonous factor. Reported mortality ranges from 8.5 to 23.2%. The disease is of significant public health concern in some localities of the world from May to November.


The Effect of Electrolytic Hydrogenation on Mechanical Properties of T92 Steel Weldments under Different PWHT Conditions.
In the present work, the effects of electrolytic hydrogen charging of T92 steel weldments on their room-temperature tensile properties were investigated. Two circumferential weldments between the T92 grade tubes were produced by gas tungsten arc welding using the matching Thermanit MTS 616 filler material. The produced weldments were individually subjected to considerably differing post-welding heat treatment (PWHT) procedures. The first-produced weldment was conventionally tempered (i.e., short-term annealed below the Ac1 critical transformation temperature of the T92 steel), whereas the second one was subjected to its full renormalization (i.e., appropriate reaustenitization well above the T92 steel Ac3 critical transformation temperature and subsequent air cooling), followed by its conventional subcritical tempering. From both weldments, cylindrical tensile specimens of cross-weld configuration were machined. The room-temperature tensile tests were performed for the individual welds' PWHT states in both hydrogen-free and electrolytically hydrogen-charged conditions. The results indicated higher hydrogen embrittlement susceptibility for the renormalized-and-tempered weldments, compared to the conventionally tempered ones. The obtained findings were correlated with performed microstructural and fractographic observations.


Controlling Pseudomonas aeruginosa persister cells by weak electrochemical currents and synergistic effects with tobramycin.
It is well recognized that bacterial populations commonly contain a small percentage of phenotypic variants, known as persister cells, which are dormant and extremely tolerant to antibiotics. When the antibiotic treatment is stopped, surviving persister cells can regenerate the bacterial population with a similar percentage of persister cells. Such persistence presents a great challenge to curing chronic infections, such as those associated with implanted medical devices. In this study, we report that bacterial persister cells can be effectively eliminated by low-level direct currents (DCs); e.g. treatment with 70 μA/cm(2) DC for 1 h using stainless steel (SS) 304 reduced the number of viable planktonic persister cells of Pseudomonas aeruginosa PAO1 by 98% compared to the untreated control. In addition to persister killing by applying DC alone, synergistic effects were observed when treating persister cells with 70 μA/cm(2) DC and 1.5 μg/mL tobramycin together using SS304 electrodes. The same level of DC was also found to be cidal to biofilms-associated persister cells of P. aeruginosa PAO1. These results are helpful for developing more effective methods to control chronic infections associated with implanted medical devices.


MglA and Igl proteins contribute to the modulation of Francisella tularensis live vaccine strain-containing phagosomes in murine macrophages.
The Francisella tularensis live vaccine strain (LVS), in contrast to its iglC mutant, replicates in the cytoplasm of macrophages. We studied the outcome of infection of the murine macrophagelike cell line J774A.1 with LVS and with iglC, iglD, and mglA mutants, the latter of which is deficient in a global regulator. Compared to LVS, all of the mutants showed impaired intracellular replication up to 72 h, and the number of the mglA mutant bacteria even decreased. Colocalization with LAMP-1 was significantly increased for all mutants compared to LVS, indicating an impaired ability to escape into the cytoplasm. A lysosomal acidity-dependent dye accumulated in approximately 40% of the vacuoles containing mutant bacteria but not at all in vacuoles containing LVS. Preactivation of the macrophages with gamma interferon inhibited the intracellular growth of all strains and significantly increased acidification of phagosomes containing the mutants, but it only slightly increased the LAMP-1 colocalization. The intracellular replication and phagosomal escape of the iglC and iglD mutants were restored by complementation in trans. In conclusion, the IglC, IglD, and MglA proteins each directly or indirectly critically contribute to the virulence of F. tularensis LVS, including its intracellular replication, cytoplasmic escape, and inhibition of acidification of the phagosomes.


Critical role for p53-serine 15 phosphorylation in stimulating transactivation at p53-responsive promoters.
The p53 tumour suppressor is induced by various stress stimuli and coordinates an adaptive gene expression programme leading to growth arrest or cell death. Some stimuli, such as DNA damage, lead to rapid and substantial multisite phosphorylation of p53, nucleated initially through phosphorylation of serine 15. Other stimuli, such as hyper-proliferation, do not stimulate p53-phosphorylation, raising questions regarding the physiological role for phosphorylation. Here, we show that a basal level of Ser15 phosphorylation occurs in both unstimulated cells and cells stimulated pharmacologically to induce p53. p53 in which Ser15 is substituted by alanine (S15A) fails to mediate p53-dependent transcription or growth arrest but can be rescued by substitution with aspartate (S15D: a phospho-mimic). Chromatin immunoprecipitation (ChIP) analyses show that, while wt- and S15A-p53 are detectable on the CDKN1A (p21) promoter (as a representative p53-responsive promoter), S15A-p53 does not stimulate histone acetylation (a measure of chromatin relaxation), nor is its recruitment stimulated, in response to a DNA damage or pharmacological stimulus. These data demonstrate that Ser15 phosphorylation is required for p53 function in the physiological context of p53-responsive promoters and suggest a key and possibly universal role even for low levels of this modification in promoting p53-transcription function.


Genomic insights into the mystery of mouse mummies on the summits of Atacama volcanoes.
Our understanding of the limits of animal life is continually revised by scientific exploration of extreme environments. Here we report the discovery of mummified cadavers of leaf-eared mice, Phyllotis vaccarum, from the summits of three different Andean volcanoes at elevations 6,029-6,233 m above sea level in the Puna de Atacama in Chile and Argentina. Such extreme elevations were previously assumed to be completely uninhabitable by mammals. In combination with a live-captured specimen of the same species from the nearby summit of Volcán Llullaillaco (6,739 m)1, the summit mummies represent the highest altitude physical records of mammals in the world. We also report a chromosome-level genome assembly for P. vaccarum that, in combination with a whole-genome re-sequencing analysis and radiocarbon dating analysis, provides insights into the provenance and antiquity of the summit mice. Radiocarbon data indicate that the most ancient of the mummies are, at most, a few centuries old. Genomic polymorphism data revealed a high degree of continuity between the summit mice and conspecifics from lower elevations in the surrounding Altiplano. Genomic data also revealed equal numbers of males and females among the summit mice and evidence of close kinship between some individuals from the same summits. These findings bolster evidence for resident populations of Phyllotis at elevations >6,000 m and challenge assumptions about the environmental limits of vertebrate life and the physiological tolerances of small mammals.


Exopolysaccharides protect Synechocystis against the deleterious effects of titanium dioxide nanoparticles in natural and artificial waters.
We have studied the effect of TiO2 nanoparticles (NPs) on the model cyanobacteria Synechocystis PCC6803. We used well-characterized NPs suspensions in artificial and natural (Seine River, France) waters. We report that NPs trigger direct (cell killing) and indirect (cell sedimentation precluding the capture of light, which is crucial to photosynthesis) deleterious effects. Both toxic effects increase with NPs concentration and are exacerbated by the presence of UVAs that increase the production of Reactive Oxygen Species (hydroxyl and superoxide radicals) by TiO2 NPs. Furthermore, we compared the responses of the wild-type strain of Synechocystis, which possesses abundant exopolysaccharides surrounding the cells, to that of an EPS-depleted mutant. We show, for the first time, that the exopolysaccharides play a crucial role in Synechocystis protection against cell killing caused by TiO2 NPs.


Glycosylation: A new signaling paradigm for the neurovascular diseases.
A wide range of life-threatening conditions with complicated pathogenesis involves neurovascular disorders encompassing Neurovascular unit (NVU) damage. The pathophysiology of NVU is characterized by several features including tissue hypoxia, stimulation of inflammatory and angiogenic processes, and the initiation of intricate molecular interactions, collectively leading to an elevation in blood-brain barrier permeability, atherosclerosis and ultimately, neurovascular diseases. The presence of compelling data about the significant involvement of the glycosylation in the development of diseases has sparked a discussion on whether the abnormal glycosylation may serve as a causal factor for neurovascular disorders, rather than being just recruited as a secondary player in regulating the critical events during the development processes like embryo growth and angiogenesis. An essential tool for both developing new anti-ischemic therapies and understanding the processes of ischemic brain damage is undertaking pre-clinical studies of neurovascular disorders. Together with the post-translational modification of proteins, the modulation of glycosylation and its enzymes implicates itself in several abnormal activities which are known to accelerate neuronal vasculopathy. Despite the failure of the majority of glycosylation-based preclinical and clinical studies over the past years, there is a significant probability to provide neuroprotection utilizing modern and advanced approaches to target abnormal glycosylation activity at embryonic stages as well. This article focuses on a variety of experimental evidence to postulate the interconnection between glycosylation and vascular disorders along with possible treatment options.


In vitro activity of Melaleuca alternifolia (tea tree) oil on filamentous fungi and toxicity to human cells.
Invasive fungal wound infections (IFIs) are increasingly reported in trauma patients and cause considerable morbidity and mortality despite standard of care treatment in trauma centers by experienced medical personnel. Topical agents such as oil of melaleuca, also known as tea tree oil (TTO), have been proposed for adjunctive treatment of IFIs. We evaluated the activity of TTO against filamentous fungi associated with IFIs by testing 13 clinical isolates representing nine species via time-kill assay with seven concentrations of TTO (100%, 75%, 50%, 25%, 10%, 5%, and 1%). To ascertain the safety of topical application to wounds, cell viability assays were performed in vitro using human fibroblasts, keratinocytes, osteoblasts, and umbilical vein endothelial cells with 10 concentrations of TTO (75%, 50%, 25%, 10%, 5%, and 10-fold serial dilutions from 1 to 0.0001%) at five time points (5, 15, 30, 60, and 180 min). Compatibility of TTO with explanted porcine tissues was also assessed with eight concentrations of TTO (100%, 75%, 50%, 25%, 10%, 5%, 1%, and 0.1%) at the time points used for cellular assays and at 24 h. The time-kill studies showed that fungicidal activity was variable between isolates. The effect of TTO on cell viability was primarily concentration dependent with significant cytotoxicity at concentrations of ≥ 10% and ≥ 50% for cells lines and whole tissue, respectively. Our findings demonstrate that TTO possesses antifungal activity against filamentous fungi associated with IFIs; furthermore that negligible effects on whole tissues, in contrast to individual cells, were observed following exposure to TTO. Collectively, these findings indicate a potential use of TTO as topical treatment of IFIs.


Multiphoton coherence domain molecular imaging with pump-probe optical coherence microscopy.
We have developed a high-resolution molecular imaging technique, pump-probe optical coherence microscopy (PPOCM), based on the fusion of pump-probe spectroscopy and optical coherence microscopy. We have demonstrated the prototype system on a fixed human skin sample containing a nodular melanoma. The results indicate that the PPOCM can clearly provide a strong contrast between the melanotic and amelanotic regions. Potential applications of the PPOCM imaging of melanin include the early diagnosis of melanoma and the mapping of tumor margins during excision. The technique may in general be applied to any biological chromophore with a known absorption spectrum.


Characterization and quantification of adeno-associated virus capsid-loading states by multi-wavelength analytical ultracentrifugation with UltraScan.
Aim: We present multi-wavelength (MW) analytical ultracentrifugation (AUC) methods offering superior accuracy for adeno-associated virus characterization and quantification. Methods: Experimental design guidelines are presented for MW sedimentation velocity and analytical buoyant density equilibrium AUC. Results: Our results were compared with dual-wavelength AUC, transmission electron microscopy and mass photometry. In contrast to dual-wavelength AUC, MW-AUC correctly quantifies adeno-associated virus capsid ratios and identifies contaminants. In contrast to transmission electron microscopy, partially filled capsids can also be detected and quantified. In contrast to mass photometry, first-principle results are obtained. Conclusion: Our study demonstrates the improved information provided by MW-AUC, highlighting the utility of several recently integrated UltraScan programs, and reinforces AUC as the gold-standard analysis for viral vectors.


Relative mass defect filtering of mass spectra: a path to discovery of plant specialized metabolites.
The rapid identification of novel plant metabolites and assignments of newly discovered substances to natural product classes present the main bottlenecks to defining plant specialized phenotypes. Although mass spectrometry provides powerful support for metabolite discovery by measuring molecular masses, ambiguities in elemental formulas often fail to reveal the biosynthetic origins of specialized metabolites detected using liquid chromatography-mass spectrometry. A promising approach for mining liquid chromatography-mass spectrometry metabolite profiling data for specific metabolite classes is achieved by calculating relative mass defects (RMDs) from molecular and fragment ions. This strategy enabled the rapid recognition of an extensive range of terpenoid metabolites in complex plant tissue extracts and is independent of retention time, abundance, and elemental formula. Using RMD filtering and tandem mass spectrometry data analysis, 24 novel elemental formulas corresponding to glycosylated sesquiterpenoid metabolites were identified in extracts of the wild tomato Solanum habrochaites LA1777 trichomes. Extensive isomerism was revealed by ultra-high-performance liquid chromatography, leading to evidence of more than 200 distinct sesquiterpenoid metabolites. RMD filtering led to the recognition of the presence of glycosides of two unusual sesquiterpenoid cores that bear limited similarity to known sesquiterpenes in the genus Solanum. In addition, RMD filtering is readily applied to existing metabolomics databases and correctly classified the annotated terpenoid metabolites in the public metabolome database for Catharanthus roseus.


Bronchodilator responsiveness in wheezy infants and toddlers is not associated with asthma risk factors.
There are limited data assessing bronchodilator responsiveness (BDR) in infants and toddlers with recurrent wheezing, and factors associated with a positive response. In a multicenter study of children ≤ 36 months old, we assessed the prevalence of and factors associated with BDR among infants/toddlers with recurrent episodes of wheezing. Forced expiratory flows and volumes using the raised-volume rapid thoracic compression method were measured in 76 infants/toddlers [mean (SD) age 16.8 (7.6) months] with recurrent wheezing before and after administration of albuterol. Prior history of hospitalization or emergency department treatment for wheezing, use of inhaled or systemic corticosteroids, physician treatment of eczema, environmental tobacco smoke exposure, and family history of asthma or allergic rhinitis were ascertained. Using the published upper limit of normal for post bronchodilator change (FEV(0.5) ≥ 13% and/or FEF(25-75) ≥ 24%) in healthy infants, 24% (n = 18) of children in our study exhibited BDR. The BDR response was not associated with any clinical factor other than body size. Dichotomizing subjects into responders (defined by published limits of normal) or by quartile to identify children with the greatest change from baseline (4th quartile vs. other) did not identify any other factor associated with BDR. Approximately one quarter of infants/toddlers with recurrent wheezing exhibited BDR at their clinical baseline. However, BDR in wheezy infants/toddlers was not associated with established clinical asthma risk factors.


Human malignancy-associated nucleolar antigen as a marker for tumor cells in patients with acute leukemia.
Tumor burden in adult patients with acute leukemia is assessed using the percentage of blast cells in the bone marrow or blood. It is clear, however, that not all blast cells are leukemic cells, especially during rapid marrow regeneration. Similarly, some leukemia cell lines have been shown to differentiate in vitro, and the same process also occurs in vivo. Therefore, the leukemic burden may be due to more differentiated cells as well as to blast cells. The purpose of this study was to investigate whether the human malignancy-associated nucleolar antigen (HMNA) could be used as a marker for leukemic cells and to examine its potential as a diagnostic tool. The proportion of HMNA-positive cells in the bone marrow of patients with acute leukemia was determined by indirect immunofluorescence with antibodies to HMNA and was compared with the differential counts routinely made in the clinic laboratory. The percentages of HMNA-positive cells among the nucleated cells in the marrow of 72 patients with clinical evidence of leukemia were significantly higher (range 9%-98%, median 83%) than those observed for nonleukemic individuals (range less than 0.05%-2.5%, median 1%) or for fractions of marrow cells from normal volunteers enriched for normal early progenitor cells (less than or equal to 2%). Patients with leukemia in remission had a lower percentage of HMNA-positive cells (range 0%-83%, median 3%). The percentage of HMNA-positive cells increased as patients approached relapse. Although the percentage of HMNA-positive cells was related to the percentage of blast cells in the bone marrow of the patients with leukemia, some partially differentiated cells were also HMNA-positive in some specimens, and some blastic cells were HMNA-negative in other specimens. These studies indicate the potential usefulness of HMNA as a marker for leukemic cells.


Printer models and error diffusion.
A new model-based approach to digital halftoning is proposed. It is intended primarily for laser printers, which generate "distortions" such as "dot overlap". Conventional methods, such as clustered-dot ordered dither, resist distortions at the expense of spatial and gray-scale resolution. The proposed approach relies on printer models that predict distortions, and rather than merely resisting them, it exploits them to increase, rather than decrease, both spatial and gray-scale resolution. We propose a general framework for printer models and find a specific model for laser printers. As an example of model-based halftoning, we propose a modification of error diffusion, which is often considered the best halftoning method for CRT displays with no significant distortions. The new version exploits the printer model to extend the benefits of error diffusion to printers. Experiments show that it provides high-quality reproductions with reasonable complexity. The proposed modified error diffusion technique is compared with Stucki's (1981) MECCA, which is a similar but not widely known technique that accounts for dot overlap. Model-based halftoning can be especially useful in transmission of high-quality documents using high-fidelity gray-scale image encoders.