Evolution of circoviruses in lorikeets lags behind its hosts.
The presence of endogenous viral elements in host genomes hints towards much older host-virus relationships than predicted by exogenous phylogenies, with highly mutable single-stranded DNA (ssDNA) viruses and RNA viruses often occupying entangled multispecies ecological niches. The difficulty lies in unravelling the long-term evolutionary history of vertebrate virus-host relationships and determining the age of a potentially ancient tree based only fresh shoots at the tips. Resolving such lineages, and the sometimes great discrepancy amongst evolutionary timescales, is problematic, especially when purifying selection or recombination can significantly alter the accuracy of phylogenetic reconstruction methods. Pathogens which occupy entangled multispecies ecological niches add a further layer of complexity but we show that multi-host scenarios may also provide opportunities to identify allopatric or sympatric paleobiological signals that can unlock longer term phylogenies. We identified host-based, cryptic, sympatric differentiation in beak and feather disease virus in the Psittaciformes tribe Loriini along with endogenous circovirus motifs in Kea (Nestor notabilis) and Gondwanan vicariance estimates to infer the evolutionary timescale of the circoviruses. This demonstrated a chronology of psittacine circovirus speciation aligned to conservative Zealandic divergences for relic circovirus motifs in Kea and a 10million year divergence coinciding with the Papuan central range orogeny that triggered the radiation of Loriini and segregation of an antecedent viral clade in Australian lorikeets. Estimates of circovirus speciation in birds highlighted a Gondwanan dominant group in Neoaves with passerine, columbid and larid circoviruses deeply separated from those in waterfowl, consistent with a Triassic divergence of Galloanserae. The circovirus tree had a deep ancestry in invertebrates with a Palaeozoic expansion in fish and mammals. We show that longer term evolutionary relationships in viruses which have a high rate of mutation and admixture can be disentangled, highlighting that contemporary virus host-switching can be explained by deep intra-lineage host phylogeny.


The future of sublingual immunotherapy.
Sublingual immunotherapy (SLIT) is currently the most prescribed form of allergen immunotherapy in many European countries. Its use has been accepted in the international consensus publications, and recently also the scepticism of USA scientists is attenuated. Still, this treatment may be improved, and the possible developments consist of modification of the materials, use of adjuvants and use of recombinant allergens. Moreover, new applications of SLIT, such as food allergy, seem promising. Concerning materials, the future form of SLIT is likely to be represented by tablets, which were already tested for efficacy and safety with grass pollen extracts, and are likely to increase the convenience for the patient by the use of no-updosing schedule. Adjuvants fitting with the characteristics of SLIT seem to be CpG oligodeoxynucleotides (CpG), able to interact with the Toll-like receptor 9 (TLR9) whose activation induces a Th1-like pattern of cytokine release, combination of 1,25-dihydroxyvitamin D3 plus dexamethasone (VitD3-Dex), and Lactobacillus plantarum. The approach with recombinant allergens, named component-resolved diagnosis, offers the possibility to tailor immunotherapy, which was found to be effective in two randomized trials of subcutaneous SIT (16-17), while studies with SLIT are not yet available. Regarding food allergy, an important controlled study demonstrated that SLIT with hazelnut is able to increase patients tolerance over possible reactions from inadvertent assumption of the culprit food, and warrants for further trials with other foods.


Exploring the genetics of lithium response in bipolar disorders.
Background: Lithium (Li) remains the treatment of choice for bipolar disorders (BP). Its mood-stabilizing effects help reduce the long-term burden of mania, depression and suicide risk in patients with BP. It also has been shown to have beneficial effects on disease-associated conditions, including sleep and cardiovascular disorders. However, the individual responses to Li treatment vary within and between diagnostic subtypes of BP (e.g. BP-I and BP-II) according to the clinical presentation. Moreover, long-term Li treatment has been linked to adverse side-effects that are a cause of concern and non-adherence, including the risk of developing chronic medical conditions such as thyroid and renal disease. In recent years, studies by the Consortium on Lithium Genetics (ConLiGen) have uncovered a number of genetic factors that contribute to the variability in Li treatment response in patients with BP. Here, we leveraged the ConLiGen cohort (N=2,064) to investigate the genetic basis of Li effects in BP. For this, we studied how Li response and linked genes associate with the psychiatric symptoms and polygenic load for medical comorbidities, placing particular emphasis on identifying differences between BP-I and BP-II. Results: We found that clinical response to Li treatment, measured with the Alda scale, was associated with a diminished burden of mania, depression, substance and alcohol abuse, psychosis and suicidal ideation in patients with BP-I and, in patients with BP-II, of depression only. Our genetic analyses showed that a stronger clinical response to Li was modestly related to lower polygenic load for diabetes and hypertension in BP-I but not BP-II. Moreover, our results suggested that a number of genes that have been previously linked to Li response variability in BP differentially relate to the psychiatric symptomatology, particularly to the numbers of manic and depressive episodes, and to the polygenic load for comorbid conditions, including diabetes, hypertension and hypothyroidism. Conclusions: Taken together, our findings suggest that the effects of Li on symptomatology and comorbidity in BP are partially modulated by common genetic factors, with differential effects between BP-I and BP-II.


SNP discovery by high-throughput sequencing in soybean.
With the advance of new massively parallel genotyping technologies, quantitative trait loci (QTL) fine mapping and map-based cloning become more achievable in identifying genes for important and complex traits. Development of high-density genetic markers in the QTL regions of specific mapping populations is essential for fine-mapping and map-based cloning of economically important genes. Single nucleotide polymorphisms (SNPs) are the most abundant form of genetic variation existing between any diverse genotypes that are usually used for QTL mapping studies. The massively parallel sequencing technologies (Roche GS/454, Illumina GA/Solexa, and ABI/SOLiD), have been widely applied to identify genome-wide sequence variations. However, it is still remains unclear whether sequence data at a low sequencing depth are enough to detect the variations existing in any QTL regions of interest in a crop genome, and how to prepare sequencing samples for a complex genome such as soybean. Therefore, with the aims of identifying SNP markers in a cost effective way for fine-mapping several QTL regions, and testing the validation rate of the putative SNPs predicted with Solexa short sequence reads at a low sequencing depth, we evaluated a pooled DNA fragment reduced representation library and SNP detection methods applied to short read sequences generated by Solexa high-throughput sequencing technology. A total of 39,022 putative SNPs were identified by the Illumina/Solexa sequencing system using a reduced representation DNA library of two parental lines of a mapping population. The validation rates of these putative SNPs predicted with low and high stringency were 72% and 85%, respectively. One hundred sixty four SNP markers resulted from the validation of putative SNPs and have been selectively chosen to target a known QTL, thereby increasing the marker density of the targeted region to one marker per 42 K bp. We have demonstrated how to quickly identify large numbers of SNPs for fine mapping of QTL regions by applying massively parallel sequencing combined with genome complexity reduction techniques. This SNP discovery approach is more efficient for targeting multiple QTL regions in a same genetic population, which can be applied to other crops.


Revisiting the (7)Li(p,n)(7)Be reaction near threshold.
In this work we review all the available experimental neutron data for the (7)Li(p,n) reaction near threshold which is necessary to obtain an accurate source model for Monte Carlo simulations in Boron Neutron Capture Therapy. Scattered published experimental results such as cross sections, differential neutron yields and total yields were collected and analyzed, exploring the sensitivity of the fitting parameters to the different possible variables and deriving a consistent working set of parameters to evaluate the neutron source near threshold.


100 Questions in Livestock Helminthology Research.
An elicitation exercise was conducted to collect and identify pressing questions concerning the study of helminths in livestock, to help guide research priorities. Questions were invited from the research community in an inclusive way. Of 385 questions submitted, 100 were chosen by online vote, with priority given to open questions in important areas that are specific enough to permit investigation within a focused project or programme of research. The final list of questions was divided into ten themes. We present the questions and set them briefly in the context of the current state of knowledge. Although subjective, the results provide a snapshot of current concerns and perceived priorities in the field of livestock helminthology, and we hope that they will stimulate ongoing or new research efforts.


Early improvement in food cravings are associated with long-term weight loss success in a large clinical sample.
Food cravings are associated with dysregulated eating behaviour and obesity, and may impede successful weight loss attempts. Gaining control over food craving is therefore a component in the management of obesity. The current paper examined whether early changes in control over food craving (assessed using the Craving Control subscale on the Control of Eating Questionnaire (CoEQ)) was predictive of weight loss in four phase 3 clinical trials investigating a sustained-release combination of naltrexone/bupropion (NB) in obese adults. The underlying component structure of the CoEQ was also examined. In an integrated analysis of four 56-week phase 3 clinical trials, subjects completed the CoEQ and had their body weight measured at baseline and at weeks 8, 16, 28 and 56. All analyses were conducted on subjects who had complete weight and CoEQ measurements at baseline and week 56, and had completed 56 weeks of NB (n=1310) or placebo (n=736). A latent growth curve model was used to examine whether early changes in the CoEQ subscales were associated with decreases in weight loss over time. Confirmatory factor analysis (CFA) was used to determine the psychometric properties of the CoEQ. The factor structure of the CoEQ was consistent with previous findings with a four-factor solution being confirmed: Craving Control, Positive Mood, Craving for Sweet and Craving for Savoury with good internal consistency (Cronbach's α=0.72-0.92). Subjects with the greatest improvement in Craving Control at week 8 exhibited a greater weight loss at week 56. These findings highlight the importance of the experience of food cravings in the treatment of obesity and support the use of the CoEQ as a psychometric tool for the measurement of food cravings in research and the pharmacological management of obesity.


Travel-associated neurological disease terminated in a postmortem diagnosed atypical HSV-1 encephalitis after high-dose steroid therapy - a case report.
Human encephalitis can originate from a variety of different aetiologies, of which infection is the most common one. The diagnostic work-up is specifically challenging in patients with travel history since a broader spectrum of unfamiliar additional infectious agents, e. g. tropical disease pathogens, needs to be considered. Here we present a case of encephalitis of unclear aetiology in a female traveller returning from Africa, who in addition developed an atypical herpes simplex virus (HSV) encephalitis in close temporal relation with high-dose steroid treatment. A previously healthy 48-year-old female presented with confusion syndrome and impaired vigilance which had developed during a six-day trip to The Gambia. The condition rapidly worsened to a comatose state. Extensive search for infectious agents including a variety of tropical disease pathogens was unsuccessful. As encephalitic signs persisted despite of calculated antimicrobial and antiviral therapy, high-dose corticosteroids were applied intravenously based on the working diagnosis of an autoimmune encephalitis. The treatment did, however, not improve the patient's condition. Four days later, bihemispheric signal amplification in the insular and frontobasal cortex was observed on magnetic resonance imaging (MRI). The intracranial pressure rapidly increased and could not be controlled by conservative treatment. The patient died due to tonsillar herniation 21 days after onset of symptoms. Histological examination of postmortem brain tissue demonstrated a generalized lymphocytic meningoencephalitis. Immunohistochemical reactions against HSV-1/2 indicated an atypical manifestation of herpesviral encephalitis in brain tissue. Moreover, HSV-1 DNA was detected by a next-generation sequencing (NGS) metagenomics approach. Retrospective analysis of cerebrospinal fluid (CSF) and serum samples revealed HSV-1 DNA only in specimens one day ante mortem. This case shows that standard high-dose steroid therapy can contribute to or possibly even trigger fulminant cerebral HSV reactivation in a critically ill patient. Thus, even if extensive laboratory diagnostics including wide-ranging search for infectious pathogens has been performed before and remained without results, continuous re-evaluation of potential differential diagnoses especially regarding opportunistic infections or reactivation of latent infections is of utmost importance, particularly if new symptoms occur.


Cell density ratios in a foveal patch in macaque retina.
We examine the assumptions that the fovea contains equal numbers of inner (invaginating or ON) and outer (flat or OFF) midget bipolar cells and equal numbers of inner and outer diffuse bipolar cells. Based on reconstruction from electron photomicrographs of serial thin sections through the fovea of a macaque monkey, we reject both assumptions. First, every foveal L and M cone is presynaptic to one inner and one outer midget bipolar cell; however, S cones are presynaptic to one outer but no inner midget bipolar cell. Second, we measure the density of all foveal cells in the same patch of fovea, affording accurate cell density ratios. For each foveal cone pedicle, at a density of 26,500 mm(-2), there is close to one (0.88) outer diffuse bipolar cell but only 0.40 inner diffuse bipolar cells. This asymmetry may be related to differences in resolution and sensitivity for light increments and decrements. We also find one (1.01) Müller cell, one (1.01) amacrine cell in the inner nuclear layer, and close to one (0.83) horizontal cell for each cone pedicle. In addition, for each S cone, there are two inner S-cone bipolar cells and two small bistratified ganglion cells. In total, there are 3.4 cone bipolar cells per cone but only 2.6 ganglion cells per cone. The latter ratio is enough to accommodate one midget ganglion cell for each midget bipolar cell.


Leveraging microenvironmental synthetic lethalities to treat cancer.
Treatment resistance leads to cancer patient mortality. Therapeutic approaches that employ synthetic lethality to target mutational vulnerabilities in key tumor cell signaling pathways have proven effective in overcoming therapeutic resistance in some cancers. Yet, tumors are organs composed of malignant cells residing within a cellular and noncellular stroma. Tumor evolution and resistance to anticancer treatment are mediated through a dynamic and reciprocal dialogue with the tumor microenvironment (TME). Accordingly, expanding tumor cell synthetic lethality to encompass contextual synthetic lethality has the potential to eradicate tumors by targeting critical TME circuits that promote tumor progression and therapeutic resistance. In this Review, we summarize current knowledge about the TME and discuss its role in treatment. We outline the concept of tumor cell-specific synthetic lethality and describe therapeutic approaches to expand this paradigm to leverage TME synthetic lethality to improve cancer therapy.


Availability of laboratory testing services for identification of periodontal pathogens in dental plaque.
Microbiologic monitoring of key subgingival plaque organisms has been proposed to enhance management of some patients with destructive forms of periodontal disease. However, isolation and identification of many of these species is available only through selected microbiology laboratories possessing special expertise with periodontal microorganisms. The purpose of this study was to determine the availability of periodontal microbiology laboratory services in the U.S. and Canada that identify suspected periodontal pathogens for dentists in clinical practice. A survey questionnaire was mailed to all U.S. and Canadian dental schools. Six laboratories that process plaque samples for dentists in private clinical practice were identified. All of the testing services examine plaque specimens for Actinobacillus actinomycetemcomitans and Bacteroides gingivalis. Four of the laboratories used bacterial culturing to characterize a wide range of plaque species and to determine antibiotic sensitivity of isolates. Two other laboratories use indirect immunofluorescence microscopy or DNA probes to screen for three putative periodontal pathogens. All but one of the testing services analyze plaque samples that are collected by private practitioners in their offices and forwarded to the laboratory through next-day delivery services or regular mail. The availability of these adjunctive diagnostic services may enhance the ability of dentists in clinical practice to apply recent advances in knowledge on periodontal microbiology into routine preventive and therapeutic patient care.


Juvenile polyposis of the stomach: clinicopathological features and its malignant potential.
To clarify a clinical entity of juvenile polyposis of the stomach compared with generalised juvenile gastrointestinal polyposis. The clinicopathological features of juvenile polyposis dominantly involving the stomach at initial presentation were reviewed in 12 patients (three new patients and nine from the literature). These were compared with 29 cases of generalised juvenile gastrointestinal polyposis. There were three men and nine women with juvenile polyposis of the stomach, aged 10-63 years. Hypoproteinaemia was present in nine patients, anaemia in seven, and a family history of intestinal polyposis in seven. No patient presented with a congenital abnormality. During the observation period, two patients developed colonic juvenile polyps. Gastric polyps invariably affected the antrum and extended to the fundus, eventually becoming more numerous, larger, and more pedunculated. Ten patients required gastrectomy for associated malignancy or uncontrolled protein losing gastropathy. Histological examinations of the resected specimens demonstrated neoplastic tissue arising from juvenile polyps in four of the 12 patients. Atypism in these mixed polyps varied from adenoma to well or moderately differentiated adenocarcinoma. Juvenile polyposis of the stomach has malignant potential, and may be a separate entity from generalised juvenile gastrointestinal polyposis.


Blood supply of pulmonary metastases.
There is a widely held belief that metastases to the lung (like primary lung tumors) are supplied by the bronchial arteries and that pulmonary neovascularization does not occur. In 17 human lungs, we have demonstrated, both in vitro and in vivo, pulmonary circulation to metastases. We have confirmed this in a series of animal studies in which we implanted tumors of various histologies into rats' tails and induced metastases from these tumors into the lungs. Forty-eight percent of these metastases had an entirely pulmonary circulation, 36% a primarily pulmonary circulation with a small bronchial component, and only 16% of metastases, located in the central one third of the lung, had an exclusively bronchial circulation. We have now shown that these supplying pulmonary vessels to metastases can be clearly demonstrated on thin-section computed tomography of the lungs. This may prove to be a helpful, noninterventional diagnostic criterion for distinguishing a primary from a metastatic lesion.


Giardia duodenalis typing from stools: a comparison of three approaches to extracting DNA, and validation of a probe-based real-time PCR typing assay.
A weighted, multi-attribute approach was used to compare three methods for direct extraction of Giardia duodenalis DNA from 15 microscopy-positive stools: (1) a QIAamp spin-column method for stools including a 10 min incubation at 95 °C, (2) method 1 preceded by five freeze-thaw cycles and (3) bead beating with guanidine thiocyanate using a FastPrep-28 machine followed by liquid-phase silica purification of DNA. The attributes compared included DNA yield measured using a new triose phosphate isomerase (tpi) gene probe-based real-time PCR, also described here. All three methods shared 100 % PCR positivity, while the bead-beating method provided the highest G. duodenalis DNA yield (P<0.01). However, when other weighted attributes, including biocontainment, resources and technical requirements, were also considered, spin-column extraction with prior freeze-thaw treatment (method 2) was deemed the most desirable and was selected for use. The tpi real-time PCR typing assay was designed to discriminate between the main human infectious assemblages of G. duodenalis (A and B) and was evaluated initially using standard isolates. Validation using microscopy-positive stools from 78 clinical giardiasis cases revealed 100 % typability; 20 (26 %) samples contained assemblage A, 56 (72 %) assemblage B and two (3 %) assemblages A and B. While the epidemiological significance of assemblage distribution will be revealed as more isolates are typed and analysed with patient demographic and exposure data, the utility of this assay and its ready application in our laboratory workflow and result turnaround margins is already evident.


[Acute mitral regurgitation caused by infarction-related papillary muscle rupture with successful surgical treatment: case report].
We present a case of severe complication of myocardial infarction -- acute mitral regurgitation caused by papillary muscle rupture. A 69-year-old man was admitted with chest pain lasting 1 hour and pulmonary oedema. ECG revealed ST-segment depression in leads II, III, aVF, V2-V6. Soon after admission the patient experienced respiratory disorders and consequently arrest. The patient was transferred in shock to the Department of Cardiothoracic Surgery, where he underwent successful artificial mitral valve implantation. One year later the patient is in good condition (NYHA class I) and the valve is fully functional.


Main group element nets to a T.
T-shaped nets (three-coordinated nets with an angular metric, bond angles near 90 degrees, 90 degrees, 180 degrees) are found in a number of extended structures. We explore in this paper the geometrical and electronic consequences of a stricter T-shape metric, where the distances between the vertices of the net are approximately equal and in the range of a chemical bond. Every atom in such a net has a T-shaped environment. One can think of these nets as extensions of BrF(3) or of substructures of various extended tellurium compounds. Several construction principles are found which allow an enumeration of a variety of one-, two-, and three-dimensional T-shape nets; not every three-coordinated net lends itself to the stricter geometrical and distance metric. Not everything is possible; there are no zero-dimensional T-nets, and none made entirely of three atom segments. Previous ideas on electron-rich multicenter bonding lead to a simple way of calculating the magic electron counts for each net; these lie in the range of 6 to 6.67.


Phylogenetic evidence for the transfer of Eubacterium lentum to the genus Eggerthella as Eggerthella lenta gen. nov., comb. nov.
Eubacterium lentum has unique phenotypic characters within the genus Eubacterium. The 16S rRNA sequence of Eubacterium lentum was determined and its phylogenetic position was defined. This micro-organism is a member of the genus Eubacterium but it is not closely related to Eubacterium limosum, the type species of the genus Eubacterium, and is nearer to Collinsella aerofaciens and Coriobacterium glomerans. A PCR-based identification system using species-specific primers designed on the basis of DNA sequences encoding the 16S rRNA of strains of Eubacterium lentum, Collinsella aerofaciens and Coriobacterium glomerans is described. A species-specific primer set can distinguish Eubacterium lentum from Eubacterium limosum or closely related species including Collinsella aerofaciens, Coriobacterium glomerans and Atopobium species. This species-specific PCR method can be used to identify Eubacterium lentum-like species isolated from human faeces. On the basis of the 16S rRNA sequence divergence from Collinsella aerofaciens and Coriobacterium glomerans and the presence of unique phenotypic characters, a new genus, Eggerthella gen. nov., is proposed for Eubacterium lentum, with one species, Eggerthella lenta comb. nov. The type strain of Eggerthella lenta is JCM 9979T.


Induction of retinoid X receptor activity and consequent upregulation of p21WAF1/CIP1 by indenoisoquinolines in MCF7 cells.
Retinoid X receptor (RXR) has been targeted for the chemoprevention and treatment of cancer. To discover potential agents acting through RXRs, we utilized an RXR response element (RXRE)-luciferase reporter gene assay. Following extensive screening, 3-amino-6-(3-aminopropyl)-5,6-dihydro-5,11-dioxo-11H-indeno[1,2-c]isoquinoline dihydrochloride (AM6-36) was found to induce RXRE-luciferase activities. AM6-36 inhibited COX-2 expression and anchorage-independent growth with 12-O-tetradecanoylphorbol 13-acetate-stimulated JB6 Cl41 cells, induced the expression of CD38 in HL-60 cells, and attenuated the growth of N-methyl-N-nitrosourea-induced mammary tumors in rats. Consistent with other reports describing the antiproliferative effects of RXR agonists in breast cancers, AM6-36 showed growth inhibition with cultured MCF7 breast cancer cells, accompanied by G(2)/M-phase arrest at lower concentrations and enhanced S-phase arrest at higher concentrations. On the basis of DNA microarray analysis, AM6-36 upregulated the expression of CDKN1A, a target gene of RXR, by 35-fold. In accord with this response, the expression of the corresponding protein, p21(WAF1/CIP1), was increased in the presence of AM6-36. Induction of p21 by AM6-36 was abrogated following transient knockdown of RXRα, demonstrating that the effect of AM6-36 on the expression of p21 is closely related to modulation of RXRα transcriptional activity. Intestinal permeability was suggested with Caco-2 cells and limited metabolism resulted when AM6-36 was incubated with human liver microsomes. Oral administration with rats resulted in 0.8 μg/mL, 4.3 μg/g, and 0.3 μg/g in serum, liver, and mammary gland, respectively. In sum, these data suggest that AM6-36 is a promising lead for the treatment or prevention of breast cancer and provide a strong rationale for testing in more advanced antitumor systems.


Artifacts in Simultaneous hdEEG/fMRI Imaging: A Nonlinear Dimensionality Reduction Approach.
Simultaneous recordings of electroencephalogram (EEG) and functional magnetic resonance imaging (fMRI) are at the forefront of technologies of interest to physicians and scientists because they combine the benefits of both modalities-better time resolution (hdEEG) and space resolution (fMRI). However, EEG measurements in the scanner contain an electromagnetic field that is induced in leads as a result of gradient switching slight head movements and vibrations, and it is corrupted by changes in the measured potential because of the Hall phenomenon. The aim of this study is to design and test a methodology for inspecting hidden EEG structures with respect to artifacts. We propose a top-down strategy to obtain additional information that is not visible in a single recording. The time-domain independent component analysis algorithm was employed to obtain independent components and spatial weights. A nonlinear dimension reduction technique t-distributed stochastic neighbor embedding was used to create low-dimensional space, which was then partitioned using the density-based spatial clustering of applications with noise (DBSCAN). The relationships between the found data structure and the used criteria were investigated. As a result, we were able to extract information from the data structure regarding electrooculographic, electrocardiographic, electromyographic and gradient artifacts. This new methodology could facilitate the identification of artifacts and their residues from simultaneous EEG in fMRI.


Intracranial pressure in patients undergoing decompressive craniectomy: new perspective on thresholds.
OBJECTIVE Decompressive craniectomy (DC) is an established part of treatment in patients suffering from malignant infarction of the middle cerebral artery (MCA) or traumatic brain injury (TBI). However, no clear evidence for intracranial pressure (ICP)-guided therapy after DC exists. The lack of this evidence might be due to the frequently used, but simplified threshold for ICP of 20 mm Hg, which determines further therapy. Therefore, the objective of this study was to evaluate this threshold's accuracy and to investigate the course of ICP values with respect to neurological outcome. METHODS Data on clinical characteristics and parameters of the ICP course on the intensive care unit were collected retrospectively in 102 patients who underwent DC between December 2007 and April 2014 at the authors' institution. The postoperative ICP course in the first 168 hours was recorded and analyzed. From these findings, ICP thresholds discriminating favorable from unfavorable outcome were calculated using conditional inference tree analysis. Additionally, survival analysis was performed using the Kaplan-Meier method. Prognostic factors were assessed via univariate analysis and multivariate logistic regression. Favorable outcome was defined as a score of 0-4 on the modified Rankin Scale. RESULTS Multivariate logistic regression revealed that anisocoria, diagnosis, and ICP values differed significantly between the outcome groups. ICP values in the favorable and unfavorable outcome groups differed significantly (p < 0.001), while the mean ICP of both groups lay below the limit of 20 mm Hg (17.5 and 11.5 mm Hg, respectively). These findings were reproduced when analyzing the underlying pathologies of TBI and MCA infarction separately. Based on these findings, optimized time-dependent threshold values were calculated and found to be between 10 and 17 mm Hg. These values significantly distinguished favorable from unfavorable outcome and predicted 30-day mortality (p < 0.001). CONCLUSIONS This study systematically evaluated ICP levels in a long-term analysis after DC and provides new, surprisingly low, time-dependent ICP thresholds for these patients. Future trials investigating the benefit of ICP-guided therapy should take these thresholds into consideration and validate them in further patient cohorts.


Translocation t(3;12)(q26;q21) in JAK2(V617F) Point Mutation Negative Chronic Idiopathic Myelofibrosis: A Case Report.
The myeloproliferative diseases (MPDs) or myelo-proliferative neoplasms (MPNs) are a group of diseases of the bone marrow in which excess cells are produced. Chronic idiopathic myelofibrosis (CIMF) is a stem cell defect characterized by splenomegaly with multiorgan extramedullary hematopoiesis, immature peripheral blood granulocytes and erythrocytes and progressive bone marrow fibrosis. The most common chromosomal abnormalities seen in CIMF patients include numerical changes of chromosomes 7, 8 and 9, and structural changes of 1q, 5q, 13q and 20q. At least 75.0% of patients with bone marrow abnormalities have one or more of these chromosomal anomalies. Detection of the Janus kinase 2 (JAK2) mutation may be a potential major breakthrough for understanding the pathobiology of MPNs, and is an essential part of the diagnostic algorithm. In this study, we describe a JAK2(V617F) mutation negative CIMF patient who has the chromosomal translocation t(3;12)(q26;q21) in her karyotype.


Image monitoring of pharmaceutical blending processes and the determination of an end point by using a portable near-infrared imaging device based on a polychromator-type near-infrared spectrometer with a high-speed and high-resolution photo diode array detector.
In the present study we have developed a new version (ND-NIRs) of a polychromator-type near-infrared (NIR) spectrometer with a high-resolution photo diode array detector, which we built before (D-NIRs). The new version has four 5 W halogen lamps compared with the three lamps for the older version. The new version also has a condenser lens with a shorter focal point length. The increase in the number of the lamps and the shortening of the focal point of the condenser lens realize high signal-to-noise ratio and high-speed NIR imaging measurement. By using the ND-NIRs we carried out the in-line monitoring of pharmaceutical blending and determined an end point of the blending process. Moreover, to determinate a more accurate end point, a NIR image of the blending sample was acquired by means of a portable NIR imaging device based on ND-NIRs. The imaging result has demonstrated that the mixing time of 8 min is enough for homogeneous mixing. In this way the present study has demonstrated that ND-NIRs and the imaging system based on a ND-NIRs hold considerable promise for process analysis.


[Results of a multicenter study of Rebif-22 mcg administration in Russia].
The results of multicenter study of Rebif 22 micrograms in Russia have been reported. 167 multiple sclerosis patients have received Rebif 22 micrograms three times a week for 1 year. This study provides evidence for reduction of the relapse rate in patients with relapsing/remitting and secondary progressive multiple sclerosis. The tendency to the decrease of the severity of relapses, less need for steroid use and the decrease of EDSS score in patients with relapsing/remitting multiple sclerosis have been shown. In general Rebif 22 micrograms was tolerated well.


Tissue-Specific Transcriptomes Reveal Gene Expression Trajectories in Two Maturing Skin Epithelial Layers in Zebrafish Embryos.
Epithelial cells are the building blocks of many organs, including skin. The vertebrate skin initially consists of two epithelial layers, the outer periderm and inner basal cell layers, which have distinct properties, functions, and fates. The embryonic periderm ultimately disappears during development, whereas basal cells proliferate to form the mature, stratified epidermis. Although much is known about mechanisms of homeostasis in mature skin, relatively little is known about the two cell types in pre-stratification skin. To define the similarities and distinctions between periderm and basal skin epithelial cells, we purified them from zebrafish at early development stages and deeply profiled their gene expression. These analyses identified groups of genes whose tissue enrichment changed at each stage, defining gene flow dynamics of maturing vertebrate epithelia. At each of 52 and 72 hr post-fertilization (hpf), more than 60% of genes enriched in skin cells were similarly expressed in both layers, indicating that they were common epithelial genes, but many others were enriched in one layer or the other. Both expected and novel genes were enriched in periderm and basal cell layers. Genes encoding extracellular matrix, junctional, cytoskeletal, and signaling proteins were prominent among those distinguishing the two epithelial cell types. In situ hybridization and BAC transgenes confirmed our expression data and provided new tools to study zebrafish skin. Collectively, these data provide a resource for studying common and distinguishing features of maturing epithelia.


Identification and characterization of microRNAs (miRNAs) and their transposable element origins in the saltwater crocodile, Crocodylus porosus.
MicroRNAs (miRNAs) are 18-24 nucleotide regulatory RNAs. They are involved in the regulation of genetic and biological pathways through post transcriptional gene silencing and/or translational repression. Data suggests a slow evolutionary rate for the saltwater crocodile (Crocodylus porosus) over the past several million years when compared to birds, the closest extant relatives of crocodilians. Understanding gene regulation in the saltwater crocodile in the context of relatively slow genomic change thus holds potential for the investigation of genomics, evolution, and adaptation. Utilizing eleven tissue types and sixteen small RNA libraries, we report 644 miRNAs in the saltwater crocodile with >78% of miRNAs being novel to crocodilians. We also identified potential targets for the miRNAs and analyzed the relationship of the miRNA repertoire to transposable elements (TEs). Results suggest an increased association of DNA transposons with miRNAs when compared to retrotransposons. This work reports the first comprehensive analysis of miRNAs in Crocodylus porosus and addresses the potential impacts of miRNAs in regulating the genome in the saltwater crocodile. In addition, the data suggests a supporting role of TEs as a source for miRNAs, adding to the increasing evidence that TEs play a significant role in the evolution of gene regulation.


Nebulized corticosteroids in the management of acute exacerbation of COPD.
Acute exacerbations in chronic onstructive pulmonary disease (COPD) are common and systemic steroids play an important role in the management of these cases along with the bronchodilators. Nebulized budesonide is being used in the acute attacks of bronchial asthma either in children or in adults. But the role of nebulized steroids in acute exacerbation of COPD is not much studied in the literature. In this clinical review we have evaluated the role of nebulized corticosteroids in the management of acute exacerbation of COPD (AECOPD). Through Medline, Pubmed and Embase we analyzed the various studies that has been done to study the role of nebulized corticosteroids in the management of acute exacerbation of COPD. The key words used for the search criteria were: acute exacerbation, COPD, nebulized corticosteroids, budesonide, fluticasone. Only eight studies were found which had evaluated the role of nebulized corticosteroids in acute exacerbations of COPD. All these studies had used nebulized budesonide in AECOPD in different dosages, and had been compared with both either parental or oral steroids, and standard bronchodilator therapy. All the studies had found the clinical efficacy of nebulized budesonide to be of similar extent to that of either parental or oral steroids in AECOPD. Side effects profile of nebulized budesonide was minimal and acceptable as compared to systemic steroids. Nebulized budesonide may be an alternative to parental/oral prednisolone in the treatment of acute exacerbations of COPD but further studies should be done to evaluate its long-term impact on clinical outcomes after an initial episode of COPD exacerbation.


Telomerase reverse transcriptase gene polymorphisms and cervical cancer susceptibility in high-risk human papillomavirus-infected women.
To investigate the relationship between Human telomerase reverse transcriptase (hTERT) gene polymorphisms and the susceptibility and clinicopathological parameters of cervical cancer in women infected with high-risk human papillomavirus (HR-HPV). A total of 380 patients with HPV-infected cervical cancer who were admitted to the Jilin province Maternal and Child Health Care Hospital (Jilin province Obstetrics Quality Control Center) from July 2019 to July 2023 were selected as case group, and 408 women with negative HPV results in the cervical cancer screening results of the physical examination in the same hospital were selected as the control group. Restriction fragment length polymorphisms polymerase chain reaction was used to detect the polymorphisms of hTERT, and its relationship with the susceptibility to high-risk HPV infection and clinicopathological parameters in patients with cervical cancer was analysed. Individuals carrying the GA and AA genotypes of rs2736122 were significantly associated with an increased risk of cervical cancer when compared with the GG genotype and the adjusted ORs were 0.53 (0.37-0.79) for the AA genotype and 0.73 (0.59-0.88) for the A allele genotype. Besides, GG genotype or G allele of rs2853677 presented a significant influence on cervical cancer, with ORs of 0.59 (0.41-0.86) and 10.77 (0.63-0.94), respectively, when compared with the AA genotype. And rs2853677 have statistically significant difference in tumour diameter and degree of differentiation subgroup(p < 0.05). The results of this study indicate that the hTERT gene rs2736122AA and rs2853677 GG genotypes can increase the susceptibility of high-risk HPV infection in cervical cancer patients. And rs2853677 is related to tumours above 4 cm and highly differentiated tumours. But both have nothing to do with the patient's chemotherapy sensitivity.


Universal total hip replacement with focus on the acetabular component and the hybrid hip.
One hundred sixty-one porous-coated Universal (Biomet Inc, Warsaw, IN) total hip arthroplasties were performed and studied preoperatively between July 1986 and June 1989. The study included 59 men (38 at 1-year follow-up; 21 at 2-year follow-up) and 91 women (60 at 1-year follow-up; 31 at 2-year follow-up). The average age was 62 years at 12-month follow-up and 64.7 years at 24-month follow-up (range 28 to 90 years). Three groups of patients with components were evaluated both clinically and radiographically: uncemented femoral and acetabular components (group 1); cemented femoral stems and uncemented acetabular components, ie, hybrid total hip arthroplasties (group 2); and revision total hip replacement arthroplasties (group 3). Clinical results for all groups showed Harris hip scores of 87 at 1-year follow-up and 89 at 2-year follow-up. Moderate to no limp occurred in 93% of patients at 1-year follow-up and 95% at 2-year follow-up. No ambulatory aids were used by 43% of patients at 1-year follow-up or 47% at 2-year follow-up. Both walking distance and endurance improved over time. Anthropometric analysis showed no progressive lucencies around the femoral or acetabular component in either group. A Kaplan-Meier survivorship analysis could not be used, because there was only one failure. This eliminated a comparison group for those lost to follow-up. Early clinical and radiographic results have been encouraging; however, long-term analysis is needed to accurately assess the success of this total hip arthroplasty system.


Prosthetic joints for the elderly.
Osteoarthritis is a consequence of other forms of arthritis, trauma, and all the uses to which we put our joints. Because it develops slowly, it is more common among elderly patients. The mechanical defects of this degenerative disease render medical treatment ineffectual. Prosthetic joints can be implanted to restore function and therefore improve the quality of life.


[Uterine sarcoma. Retrospective analysis of a series of 22 cases].
37 uterine tumors originally diagnosed as uterine sarcomas at Fondation Bergonié between 1970 and 1982 were histologically reclassified. 15 patients were excluded from this study because of inadequate data or not confirmed initial diagnosis. The 22 remaining cases were classified as leiomyosarcomas 13, endometrial stromal sarcomas 7 and mixed mesodermal sarcomas 2. The mean age of the patients was 56. The cases were classified according to the FIGO staging system. 5 patients had previously undergone (between 1 and 8 years) a subtotal hysterectomy for uterine leiomyoma. 5 patients had stage 1 sarcoma, 13 patients stage III and 2 patients stage IV. 13 patients are dead, 7 are alive without evidence of disease, 2 are alive with evidence of disease. The 5 year actuarial survival rate was 30%. Tumor extent at diagnosis was not correlated with survival. Analysis of failures revealed : 4 metastatic, 5 metastatic and pelvic and 6 isolated pelvic failures. Prognosis was not significantly different between leiomyosarcomas and endometrial stromal sarcomas. There was no pelvic failure in patients with stage I sarcoma (treated by surgery plus radiotherapy). Survival was not found to be correlated with histopathological grade as described in soft tissue sarcomas. Nevertheless a correlation (non significant) was found between survival and mitosis count. Chemotherapy (Cyvadic) was administered in 8 advanced tumors with partial response (always below 50% of the initial tumor) in 5 patients.


A tunable delivery platform to provide local chemotherapy for pancreatic ductal adenocarcinoma.
Pancreatic ductal adenocarcinoma (PDAC) is one of the most devastating and painful cancers. It is often highly resistant to therapy owing to inherent chemoresistance and the desmoplastic response that creates a barrier of fibrous tissue preventing transport of chemotherapeutics into the tumor. The growth of the tumor in pancreatic cancer often leads to invasion of other organs and partial or complete biliary obstruction, inducing intense pain for patients and necessitating tumor resection or repeated stenting. Here, we have developed a delivery device to provide enhanced palliative therapy for pancreatic cancer patients by providing high concentrations of chemotherapeutic compounds locally at the tumor site. This treatment could reduce the need for repeated procedures in advanced PDAC patients to debulk the tumor mass or stent the obstructed bile duct. To facilitate clinical translation, we created the device out of currently approved materials and drugs. We engineered an implantable poly(lactic-co-glycolic)-based biodegradable device that is able to linearly release high doses of chemotherapeutic drugs for up to 60 days. We created five patient-derived PDAC cell lines and tested their sensitivity to approved chemotherapeutic compounds. These in vitro experiments showed that paclitaxel was the most effective single agent across all cell lines. We compared the efficacy of systemic and local paclitaxel therapy on the patient-derived cell lines in an orthotopic xenograft model in mice (PDX). In this model, we found up to a 12-fold increase in suppression of tumor growth by local therapy in comparison to systemic administration and reduce retention into off-target organs. Herein, we highlight the efficacy of a local therapeutic approach to overcome PDAC chemoresistance and reduce the need for repeated interventions and biliary obstruction by preventing local tumor growth. Our results underscore the urgent need for an implantable drug-eluting platform to deliver cytotoxic agents directly within the tumor mass as a novel therapeutic strategy for patients with pancreatic cancer.


Online Education and Cognitive Behavior Therapy Improve Dementia Caregivers' Mental Health: A Randomized Trial.
To compare online cognitive-behavioral therapy (CBT) with and without telephone support respectively to online psychoeducation in a randomized controlled trial (RCT) in caregivers of people with dementia with mild anxiety or depression. Three-arm parallel-group RCT comparing online CBT with and without telephone support respectively to online psychoeducation. Online study with caregivers of people with dementia. The primary outcome measure was mental health measured by General Health Questionnaire-12 (GHQ-12) at 26 weeks. Secondary outcomes included the Hospital Anxiety and Depression Scale (HADS); the Relative Stress Scale (RSS) and the Short Sense of Competency Questionnaire. The primary analysis focused on people completing GHQ-12 at both baseline and 26 weeks, evaluated using analysis of covariance. 638 people were randomized to the 3 treatment arms, of whom 208 were included in the analysis population. There were significant improvements in GHQ-12 in all treatment arms compared to baseline (P < .001 for all interventions), but neither CBT with nor without telephone support conferred any significant advantage compared to psychoeducation. For the secondary outcomes, there were no significant differences between CBT with telephone support and psychoeducation, but CBT without telephone support was less effective than psychoeducation with respect to HADS depression subscale [mean difference 1.86, 95% confidence interval (CI) 0.61, 3.11; P = .004] and caregiver stress (RSS mean difference 3.11, 95% CI 0.13, 6.09; P = .04). Good safety was achieved in all 3 treatment arms, with no deaths or serious adverse events. Online CBT with telephone support and psychoeducation both achieved significant benefits over 26 weeks compared with baseline in mental health and mood, but there were no advantages for CBT compared with the psychoeducation intervention. CBT without telephone support was less effective with respect to mood outcomes than psychoeducation and should not be recommended based on current evidence.


Laser hair removal: a review and report on the use of the long-pulsed alexandrite laser for hair reduction of the upper lip, leg, back, and bikini region.
The mechanism and permanence of laser-assisted hair removal remains a formidable task in the medical community. The purpose of this study was to determine the safety and long-term efficacy of the long-pulsed or normal mode alexandrite infrared laser for hair depilation. Beginning in October 1996, a total of 31 anatomic sites on 22 patients ranging in age from 25 to 59 years (mean 42 years) were evaluated to assess hair removal. Treatment sites included 17 upper lips, 9 legs, 2 backs, and 3 bikini regions. Eligible patients were of Fitzpatrick skin types I-III. Patients were treated using the long-pulsed alexandrite infrared laser at 755 nm, single-pulse technique, 10 mm spot size, 10% overlap, pulse durations of 5, 10, and 20 msec, and a fluence of 20 J/cm2. Subjective patient improvement and objective, blinded graded improvement was assessed at 1, 2, 3, and 6 months. Objective blinded grading at 6 months revealed that hair reduction varied both with the pulse duration and anatomic location. Maximum reductions observed were 40%, 56%, 50%, and 15% for the lip, leg, back, and bikini areas, respectively. Upper lip hair reduction increased from 40% to 54% at 6 months when a second treatment was performed 8 weeks after the initial treatment. The long-pulsed alexandrite laser is safe and effective in reducing hair growth. Treatment efficacy varies with the anatomic location, pulse duration, and number of treatments. A single-pulse technique utilizing a 10 msec pulse duration at 20 J/cm2 produced the greatest hair reduction. No permanent adverse effects occurred on skin types I-III at the parameters tested.


[Analysis of efficacy and prognosis of neoadjuvant chemotherapy and (or) surgery plus radiotherapy for hypopharyngeal squamous cell carcinoma].
Objective:To analyze the risk factors that affect the prognosis of patients with hypopharyngeal squamous cell carcinoma(HPSCC) and to compare the efficacy of surgical resection followed by adjuvant radiotherapy(SR) with that of neoadjuvant therapy consisting of platinum-based chemotherapy and fluorouracil combined with either cetuximab or nimotuzumab, followed by SR. The study also aimed to evaluate the overall survival(OS) of patients, their postoperative eating function, tracheostomy decannulation rate, and tumor response to the two neoadjuvant chemotherapies. Methods:A retrospective analysis was performed on the medical records of HPSCC patients who received SR or neoadjuvant therapy followed by SR treatment at the Shanghai General Hospital from 2012 to 2019 and had not undergone any prior treatment. The prognostic factors were analyzed, and the survival analysis of patients who underwent SR treatment with two neoadjuvant chemotherapy regimens was performed. Results:A total of 108 patients were included in the study. The results of the univariate analysis showed that gender(P=0.850) had no significant correlation with the survival rate of HPSCC patients who underwent SR. However, age, smoking history, alcohol consumption history, platelet-to-lymphocyte ratio(PLR), neutrophil-to-lymphocyte ratio(NLR), T stage, N stage, neoadjuvant therapy with either cetuximab or nimotuzumab combined with platinum-based chemotherapy and fluorouracil, and histological grade were significantly associated with prognosis(P<0.05). The multivariate analysis revealed that smoking history, histological grade, and neoadjuvant therapy with either cetuximab or nimotuzumab combined with platinum-based chemotherapy and fluorouracil were independent risk factors affecting the prognosis of HPSCC(P<0.05). Patients who received neoadjuvant therapy had longer OS than those who underwent SR only(P<0.001). There was no significant difference in tumor response to the two neoadjuvant therapies and in OS(P>0.05), and there was no significant difference in the rate of oral feeding and tracheostomy decannulation among the three treatment groups(P>0.05). Conclusion:Univariate analysis showed that age at tumor onset, smoking history, alcohol consumption history, NLR, PLR, T stage, N stage, whether receiving neoadjuvant chemotherapy, and pathological grade were associated with the prognosis of HPSCC patients receiving SR treatment. Multivariate analysis showed that smoking history, pathological grade, and neoadjuvant chemotherapy were independent risk factors affecting the prognosis. Neoadjuvant chemotherapy with cetuximab or nimotuzumab can prolong the OS of patients, providing a certain basis and reference for the treatment of HPSCC.


Wideband four-level transmission gratings with flattened spectral efficiency.
Four-level transmission-type surface relief grating profiles with nearly flat efficiency over a spectral octave are designed by rigorous electromagnetic diffraction theory. Parametric optimization of the relief depths and transition points of the profile steps of these leads to efficiencies in the range 50-60% over the entire octave if the ratio of the grating period and the mean spectral wavelength is greater than ~ 3.


[Brucellosis-induced granulomatous non-puerperal mastitis--a case report].
An acute manifestation of a chronic brucellosis localised to the female breast is described. A granulomatous mastitis was identified on histological examination. To our knowledge, this is the first reported occurrence of this phenomenon. It is pointed out that it is clinically difficult to distinguish this type of infection from an inflammatory carcinoma of the breast.


Long non-coding RNA (lncRNA) MALAT1 in regulating osteogenic and adipogenic differentiation using a double-stranded gapmer locked nucleic acid nanobiosensor.
Long non-coding RNAs (lncRNA) are non-protein coding RNA molecules that are longer than 200 nucleotides. The lncRNA molecule plays diverse roles in gene regulation, chromatin remodeling, and cellular processes, influencing various biological pathways. However, probing the complex dynamics of lncRNA in live cells is a challenging task. In this study, a double-stranded gapmer locked nucleic acid (ds-GapM-LNA) nanobiosensor is designed for visualizing the abundance and expression of lncRNA in live human bone-marrow-derived mesenchymal stem cells (hMSCs). The sensitivity, specificity, and stability were characterized. The results showed that this ds-GapM-LNA nanobiosensor has very good sensitivity, specificity, and stability, which allows for dissecting the regulatory roles of cellular processes during dynamic physiological events. By incorporating this nanobiosensor in living hMSC imaging, we elucidated lncRNA MALAT1 expression dynamics during osteogenic and adipogenic differentiation. The data reveal that lncRNA MALAT1 expression is correlated with distinct sub-stages of osteogenic and adipogenic differentiation.


MOG-Ab prevalence in optic neuritis and clinical predictive factors for diagnosis.
What is the proportion of antibodies to myelin oligodendrocyte glycoprotein (MOG-Ab) in optic neuritis (ON) in adults and what would be the ON presentation for which MOG-Ab should be tested? Multicentric prospective study conducted during 1 year on all patients diagnosed with acute ON in all ophthalmological units in hospitals in a region in western France. Sixty-five patients were included. MOG-Ab prevalence was 14% (9/65) during an acute ON and 13% (7/55) after exclusion of patients already diagnosed with multiple sclerosis (MS) (8) or MOG+ON (2). Compared with MS and clinically isolated syndrome, MOG+ON had no female preponderance (67% of men in case of MOG+ON and 22% of men in case of MS and clinically isolated syndrome, p<0.05) were more often bilateral (44% vs 3%, p<0.005) and associated with optic disc swelling (ODS) (78% vs 14%, p<0.001). To predict MOG+ON, the positive predictive values (PPVs) of male sex, ODS and bilateral involvement were 29% (95% CI 9% to 48%), 41% (95% CI 18% to 65%) and 40% (95% CI 10% to 70%), respectively, while the negative predictive values (NPV) were 93% (95% CI 86% to 100%), 96% (95% CI 90% to 100%) and 91% (95% CI 83% to 99%), respectively. The combined factor 'ODS or bilateral or recurrent ON' was the best compromise between PPV (31% (95% CI 14% to 48%)) and NPV (100% (95% CI 100% to 100%)). Among ON episodes, MOG-Ab were found in 14% of cases. MOG+ON occurred without female preponderance and was significantly associated with ODS and/or bilateral ON. Testing MOG-Ab only in patients presenting with ODS or bilateral or recurrent ON would limit MOG-Ab tests to fewer than half of all patients without the risk of missing any MOG+ON cases.


Inhibition of Protease and Egg Hatching of Haemonchus contortus by Soybean Seed Exudates.
Gastrointestinal nematode infection of small ruminants causes losses in livestock production. Plant compounds show promises as alternatives to commercial anthelmintics that have been exerting selective pressures that lead to the development of drug-resistant parasites. Soybean (Glycine max) is an economical value crop, with a higher protein content compared to other legumes. The objective of this study was to evaluate whether the protease inhibitors exuded from the G. max mature seeds have anthelmintic activity against Haemonchus contortus. To obtain the soybean exudates (SEX), mature seeds were immersed in 100 mM sodium acetate buffer, pH 5.0, at 10 C, for 24 hr. Then the naturally released substances present in SEX were collected and exhaustively dialyzed (cutoff 12 kDa) against distilled water. The dialyzed seed exudates (SEXD) were heated at 100 C for 10 min and centrifuged (12,000 g, at 4 C for 15 min). The supernatant obtained was recovered and designated as the heat-treated exudate fraction (SEXDH). The protein content, protease inhibitor activity, and the effect of each fraction on H. contortus egg hatch rate were evaluated. The inhibition extent of SEX, SEXD, and SEXDH on H. contortus egg proteases was 31.1, 42.9, and 63.8%, respectively. Moreover, SEX, SEXD, and SEXDH inhibited the egg hatching with EC50 of 0.175, 0.175, and 0.241 mg ml-1, respectively. Among the commercial protease inhibitors tested, only EDTA and E-64 inhibited the H. contortus hatch rate (79.0 and 28.9%, respectively). We present evidence demonstrating that soybean exudate proteins can effectively inhibit H. contortus egg hatching. This bioactivity is displayed by thermostable proteins and provides evidence that protease inhibitors are a potential candidate for anthelmintic use.


Dietary omega-3 fatty acid supplementation increases the rate of muscle protein synthesis in older adults: a randomized controlled trial.
Loss of muscle mass with aging is a major public health concern. Omega-3 (n-3) fatty acids stimulate protein anabolism in animals and might therefore be useful for the treatment of sarcopenia. However, the effect of omega-3 fatty acids on human protein metabolism is unknown. The objective of this study was to evaluate the effect of omega-3 fatty acid supplementation on the rate of muscle protein synthesis in older adults. Sixteen healthy, older adults were randomly assigned to receive either omega-3 fatty acids or corn oil for 8 wk. The rate of muscle protein synthesis and the phosphorylation of key elements of the anabolic signaling pathway were evaluated before and after supplementation during basal, postabsorptive conditions and during a hyperaminoacidemic-hyperinsulinemic clamp. Corn oil supplementation had no effect on the muscle protein synthesis rate and the extent of anabolic signaling element phosphorylation in muscle. Omega-3 fatty acid supplementation had no effect on the basal rate of muscle protein synthesis (mean ± SEM: 0.051 ± 0.005%/h compared with 0.053 ± 0.008%/h before and after supplementation, respectively; P = 0.80) but augmented the hyperaminoacidemia-hyperinsulinemia-induced increase in the rate of muscle protein synthesis (from 0.009 ± 0.005%/h above basal values to 0.031 ± 0.003%/h above basal values; P < 0.01), which was accompanied by greater increases in muscle mTOR(Ser2448) (P = 0.08) and p70s6k(Thr389) (P < 0.01) phosphorylation. Omega-3 fatty acids stimulate muscle protein synthesis in older adults and may be useful for the prevention and treatment of sarcopenia. This trial was registered at clinical trials.gov as NCT00794079.


Studies on osteoporosis III. Effect of estrogens and fluoride.
Total skeletal calcium levels were determined in female mice with the aid of whole body neutron activation analysis. Three months treatment with heparin produced significant osteoporosis in C3-H/St(Ha) mice but not in C57/BL6 (J) mice. Treatment with a conjugated natural estrogen preparation (Premarin) prevented heparin accelerated osteoporosis, but high level fluoride in the drinking water had no preventive effect. In some experiments there was a suggestion of a deleterious effect of fluoride.


Recurrence after topical nonpreserved methylprednisolone therapy for keratoconjunctivitis sicca in Sjögren's syndrome.
The aim of this study was to evaluate the prevalence of long-term recurrence after topical nonpreserved methylprednisolone pulse therapy for the treatment of keratoconjunctivitis sicca (KCS) with Sjögren's syndrome. A total of 106 eyes of 53 Sjögren's syndrome patients were included in the study. All patients were treated with topical nonpreserved 1% methylprednisolone solution. Initial therapy consisted of eyedrops 4 times a day for 2 weeks, and then patients were reevaluated and tapered off the medication every 2 weeks until discontinuation. Tear film breakup time (TBUT), Schirmer test, corneal fluorescein staining, and subjective symptom scores were measured. Additionally, impression cytology of the bulbar conjunctiva was performed. The over-all success rates and success period were determined using the Kaplan-Meier survival curve. A reduction in subjective symptoms and fluorescein staining, and an improvement in TBUT and Schirmer test results, was observed after treatment (P<0.001). Impression cytology specimens revealed a significantly increased number of periodic acid-Schiff-positive cells after treatment. After the first pulse therapy, mean survival was 56.6 weeks and 11 (20.8%) patients recurred. After the second pulse therapy, mean survival was 72.4 weeks and only 1 patient recurred. No serious complications, including intraocular pressure elevation and cataract formation, were encountered during the entire follow-up period. Topical nonpreserved methylprednisolone pulse therapy proved to be a safe, effective long-term treatment of improving subjective and objective dry eye factors in KCS patients with Sjögren's syndrome.


Assessing the impact of mass media public health campaigns. Be Clear on Cancer 'blood in pee': a case in point.
To assess the impact on suspected cancer referral burden and new cancer diagnosis of Public Health England's recent Be Clear on Cancer 'blood in pee' mass media campaign. A retrospective cohort study design was used. For two distinct time periods, August 2012 to May 2013 and August 2013 to May 2014, all referrals of patients deemed to be at risk of urological cancer by the referring primary healthcare physician to Imperial College NHS Healthcare Trust were screened. Data were collected on age and sex and whether the referral was for visible haematuria, non-visible haematuria or other suspected urological cancer. In addition to referral data, hospital episode data for all new renal cell (RCC) and upper and lower tract transitional cell carcinoma (TCC), as well as testicular and prostate cancer diagnoses for the same time periods were obtained. Over the campaign period and the subsequent 3 months, the number of haematuria referrals increased by 92% (P = 0.013) when compared with the same period a year earlier. This increase in referrals was not associated with a significant corresponding rise in cancer diagnosis; instead changes of 26.8% (P = 0.56) and -3.3% (P = 0.84) were seen in RCC and TCC, respectively. This study has shown that the Be Clear on Cancer 'blood in pee' mass media campaign significantly increased the number of new suspected cancer referrals, but there was no significant change in the diagnosis of target cancers across a large catchment. Mass media campaigns are expensive, require significant planning and appropriate implementation and, while the findings of this study do not challenge their fundamental objective, more work needs to be done to understand why no significant change in target cancers was observed. Further consideration should also be given to the increased referral burden that results from these campaigns, such that pre-emptive strategies, including educational and process mapping, across primary and secondary care can be implemented.


State-Level Patterns and Trends in Cigarette Smoking Across Racial and Ethnic Groups in the United States, 2011-2018.
Reducing racial/ethnic disparities in smoking is a priority for state tobacco control programs. We investigated disparities in cigarette use by race/ethnicity, as well as trends in cigarette use across racial/ethnic groups from 2011 to 2018 in 50 US states and the District of Columbia. We used data from the Behavioral Risk Factor Surveillance System. In each state, smoking prevalence and corresponding 95% CIs were estimated for each racial/ethnic group in 2011, 2014, and 2018. We used logistic regression models to examine state-specific linear and quadratic time trends in smoking prevalence from 2011 to 2018. Racial/ethnic disparities in smoking prevalence varied across states. From 2011 to 2018, compared with White adults, the odds of smoking were lower among Black adults in 14 states (odds ratio [OR] range, 0.58-0.91) and were higher in 9 states (OR range, 1.10-1.98); no differences were found in the odds of smoking in 13 states. Compared with White adults, the odds of smoking were lower among Hispanic adults in most states (OR range, 0.33-0.84) and were typically higher among Other adults (OR range, 1.19-2.44). Significant interactions between year and race/ethnicity were found in 4 states, indicating that time trends varied across racial/ethnic groups. In states with differential time trends, the decline in the odds of smoking was typically greater among Black, Hispanic, and Other adults compared with White adults. Some progress in reducing racial/ethnic disparities in smoking has been made, but additional efforts are needed to eliminate racial/ethnic disparities in smoking.


[Economic evaluation of pharmacotherapeutic follow-up in type 2 diabetes mellitus patients in community pharmacies].
Assess economics results of Pharmacotherapeutic Follow-up (PF) in patients with diabetes mellitus type 2 in community pharmacies from additional Health system. In a prospective clinic study, 161 patients were divided into two groups, of which only one group received PF for 12 months. From the results, we calculated data of effectiveness and costs. The primary endpoint was the economic evaluation PF by using an indicator of effectiveness (changes in glycated hemoglobin), which was related to the costs of pharmaceutical care. Secondary endpoints were values of blood pressure, waist circumference and body mass index of patients, also related to costs of pharmaceutical care. A real reduction of 1.3% of HbA1 was observed in the PF group, in comparison to control group. The annual cost of the reduction in 1% in HbA1 values in the PF group patients was $45.15. This feature to improve the control of type 2 diabetic patients can be implemented with annual investments of about $225.76 per patient, using PF and monitoring of blood glucose test. It is possible to reduce the HbA1 values to desired levels by using PF. This can be considered an additional resource for the attainment of metabolic control, resulting in this study a cost of $37.62 per patient, per year, to reduce 1% in the HbA1 values.


Better-surviving liver grafts by the injection of anti-CD2 antibody: the important roles of host CD8+ and CD2+CD28+ T cells in chronic graft rejection and beta type platelet-derived growth factor receptor (PDGFR-beta) expression on apoptotic liver grafts.
Syngeneic liver grafts were implanted in the livers of 22 LEW/Sea strain rats. To prolong the graft survival, anti-CD2 monoclonal antibody (MAb) or anti beta type platelet-derived growth factor receptor (PDGFR-beta) antibody (Ab) was injected, or splenectomy was performed in the rats which were then followed until 10 to 11 weeks posttransplantation. The 22 rats with chronic graft rejection showed increased CD8a-like antigen (probably Fas ligand) on the peripheral blood T cells. All the liver grafts had both necrosis and apoptosis. The liver graft apoptosis was indicated by histopathological abnormalities, and by DNA strand breaks and hemosiderin depositions in the cytoplasm. PDGFR-beta expression in the apoptotic liver graft was demonstrated immunohistochemically. Among the 17 rats injected with anti-CD2 MAb, CD2 signaling on host T cells was effectively suppressed by the injection of anti-CD2 MAb in 4 rats with better-surviving liver grafts. In these 4 rats, CD28 antigen on thymic lymphocytes was down-modulated and high numbers (136-233-positive cells per lobe) of the epithelial reticular cells with apoptotic lymphocytes were counted. Anti-PDGFR-beta Ab caused high pulmonary secretions of growth factors and reticular fibrosis in the lungs of 5 rats injected with the Ab. Anti-PDGFR-beta Ab injection reduced the host cell apoptosis in the lung and thymus, but did not prolong the survival of liver grafts. In the 9 rats with both splenectomy and anti-CD2 MAb injection, pulmonary apoptosis was induced with the 6-16% reductions of CD4+ lymphocytes. Prolonged graft survival was observed in only one of the 9 rats. Anti-CD2 MAb was effective for prolonging the liver graft survival with suppressed CD28 antigen, but anti-PDGFR-beta Ab and splenectomy were not.


Measuring patient-centered medical home access and continuity in clinics with part-time clinicians.
Common patient-centered medical home (PCMH) performance measures value access to a single primary care provider (PCP), which may have unintended consequences for clinics that rely on part-time PCPs and team-based care. Retrospective analysis of 110,454 primary care visits from 2 Veterans Health Administration clinics from 2010 to 2012. Multi-level models examined associations between PCP availability in clinic, and performance on access and continuity measures. Patient experiences with access and continuity were compared using 2012 patient survey data (N = 2881). Patients of PCPs with fewer half-day clinic sessions per week were significantly less likely to get a requested same-day appointment with their usual PCP (predicted probability 17% for PCPs with 2 sessions/week, 20% for 5 sessions/week, and 26% for 10 sessions/week). Among requests that did not result in a same-day appointment with the usual PCP, there were no significant differences in same-day access to a different PCP, or access within 2 to 7 days with patients' usual PCP. Overall, patients had >92% continuity with their usual PCP at the hospital-based site regardless of PCP sessions/week. Patients of full-time PCPs reported timely appointments for urgent needs more often than patients of part-time PCPs (82% vs 71%; P < .01), but reported similar experiences with routine access and continuity. Part-time PCP performance appeared worse when using measures focused on same-day access to patients' usual PCP. However, clinic-level same-day access, same-week access to the usual PCP, and overall continuity were similar for patients of part-time and full-time PCPs. Measures of in-person access to a usual PCP do not capture alternate access approaches encouraged by PCMH, and often used by part-time providers, such as team-based or non-face-to-face care.


QAGView: Interactively Summarizing High-Valued Aggregate Query Answers.
Methods for summarizing and diversifying query results have drawn significant attention recently, because they help present query results with lots of tuples to users in more informative ways. We present QAGView (Quick AGgregate View), which provides a holistic overview of high-valued aggregate query answers to the user in the form of summaries (showing high-level properties that emerge from subsets of answers) with coverage guarantee (for a user-specified number of top-valued answers) that is both diverse (avoiding overlapping or similar summaries) and relevant (focusing on high-valued aggregate answers). QAGView allows users to view the high-level summaries as clusters, and to expand individual clusters for their constituent result tuples. Users can fine-tune the behavior of QAGView by specifying a number of parameters according their preference. To help users choose appropriate parameters interactively, QAGView employ a suite of optimizations that enable quick preview of how the quality of the summaries changes over wide ranges of parameter settings, as well as real-time visualization of how the summaries evolve in response to parameter updates.


Over 65% Sunlight Absorption in a 1 μm Si Slab with Hyperuniform Texture.
Thin, flexible, and invisible solar cells will be a ubiquitous technology in the near future. Ultrathin crystalline silicon (c-Si) cells capitalize on the success of bulk silicon cells while being lightweight and mechanically flexible, but suffer from poor absorption and efficiency. Here we present a new family of surface texturing, based on correlated disordered hyperuniform patterns, capable of efficiently coupling the incident spectrum into the silicon slab optical modes. We experimentally demonstrate 66.5% solar light absorption in free-standing 1 μm c-Si layers by hyperuniform nanostructuring for the spectral range of 400 to 1050 nm. The absorption equivalent photocurrent derived from our measurements is 26.3 mA/cm2, which is far above the highest found in literature for Si of similar thickness. Considering state-of-the-art Si PV technologies, we estimate that the enhanced light trapping can result in a cell efficiency above 15%. The light absorption can potentially be increased up to 33.8 mA/cm2 by incorporating a back-reflector and improved antireflection, for which we estimate a photovoltaic efficiency above 21% for 1 μm thick Si cells.


Confocal microscopic analysis of intracellular pH regulation in isolated guinea pig pancreatic ducts.
pH regulation in isolated guinea pig pancreatic interlobular duct segments loaded with the pH-sensitive fluorophore, 5-(6)-carboxy-SNARF-1-acetoxymethyl ester (SNARF-1), was characterized by laser-scanning confocal microscopy. In HCO3(-)-free medium, intracellular pH (pHi) of duct epithelial cells fell by 0.32 +/- 0.06 pH units in the presence of 0.5 mM amiloride and by 0.36 +/- 0.08 pH units in the absence of Na+. In the presence of extracellular HCO3-, pHi acidified in Na(-)-free medium but not in amiloride-containing medium. Superfusion with Cl(-)-free buffers or with buffers containing 4,4'-diisothiocyanostilbene-2,2'-disulfonic acid produced a cytosolic alkalinization of 0.13-0.22 pH units. These observations demonstrate the presence of Na+/H+ exchange, Na(+)-HCO3- cotransport, and Cl-/HCO3- exchange in guinea pig pancreatic ducts. pHi recovered significantly from an NH4Cl pulse in HCO3(-)-free buffers containing amiloride and carbachol (50.4%) or amiloride and secretin (40.6%). This recovery was blocked by the H(+)-adenosinetriphosphatase (H(+)-ATPase) inhibitor bafilomycin A1 and by preincubation of ducts with nocodazole or cytochalasin D. These observations suggest that a vesicular H(+)-ATPase augments Na(+)-dependent H+ extrusion during agonist-stimulated bicarbonate secretion and that activation of this transport mechanism involves cytoskeletal elements.


The prevalence of infections with Trichophyton tonsurans in schoolchildren: the CAPITIS study.
Although Trichophyton tonsurans has become the leading cause of tinea capitis in the United States, reported infection rates vary widely, and prevalence estimates for the pediatric population at large remain poorly characterized. A prospective, cross-sectional, surveillance study of children attending kindergarten through fifth grade in 44 schools across the bi-state (Kansas/Missouri), Kansas City metropolitan area was conducted. Fungal cultures were collected from all participants, and molecular analyses were used to characterize the patterns of infection within the population. Of 10,514 children (age: 8.3 +/- 1.9 years) examined for the presence of T tonsurans on their scalps, 6.6% exhibited positive cultures. Infection rates at participating schools ranged from 0% to 19.4%, exceeding 30% at a given grade level in some schools. Black children demonstrated the highest rates of infection (12.9%), with prevalence estimates for the youngest members of this racial group approaching 18%. Infection rates for Hispanic (1.6%) and white (1.1%) children were markedly lower. A single genetic strain of T tonsurans was identified in only 16.6% of classrooms, whereas each child harbored a unique genetic strain in 51.4%. We report a large-scale, citywide, surveillance study of T tonsurans infection rates among children in primary school in a metropolitan area. The striking prevalence rates and genetic heterogeneity among the fungal isolates confirm the relatively large degree to which this pathogen has become integrated into metropolitan communities.


Anthrax lethal toxin disrupts intestinal barrier function and causes systemic infections with enteric bacteria.
A variety of intestinal pathogens have virulence factors that target mitogen activated protein kinase (MAPK) signaling pathways, including Bacillus anthracis. Anthrax lethal toxin (LT) has specific proteolytic activity against the upstream regulators of MAPKs, the MAPK kinases (MKKs). Using a murine model of intoxication, we show that LT causes the dose-dependent disruption of intestinal epithelial integrity, characterized by mucosal erosion, ulceration, and bleeding. This pathology correlates with an LT-dependent blockade of intestinal crypt cell proliferation, accompanied by marked apoptosis in the villus tips. C57BL/6J mice treated with intravenous LT nearly uniformly develop systemic infections with commensal enteric organisms within 72 hours of administration. LT-dependent intestinal pathology depends upon its proteolytic activity and is partially attenuated by co-administration of broad spectrum antibiotics, indicating that it is both a cause and an effect of infection. These findings indicate that targeting of MAPK signaling pathways by anthrax LT compromises the structural integrity of the mucosal layer, serving to undermine the effectiveness of the intestinal barrier. Combined with the well-described immunosuppressive effects of LT, this disruption of the intestinal barrier provides a potential mechanism for host invasion via the enteric route, a common portal of entry during the natural infection cycle of Bacillus anthracis.


Cryptococcus gattii Meningitis in a Previously Healthy Young Woman: A Case Report.
Cryptococcus gattii (C. gatti) is a rare cause of meningitis in the United States. Outbreaks in new geographic distributions in the past few decades raise concern that climate change may be contributing to a broader distribution of this pathogen. We review a case of C. gattii in a 23-year-old woman in Northern California who was diagnosed via lumbar puncture after six weeks of headache, blurred vision, and tinnitus. A 23-year-old previously healthy young woman presented to the emergency department (ED) after multiple visits to primary care, other EDs, and neurologists, for several weeks of headache, nausea, tinnitus, and blurred vision. On examination the patient was found to have a cranial nerve VI palsy (impaired abduction of the left eye) and bilateral papilledema on exam. Lumbar puncture had a significantly elevated opening pressure. Cerebrospinal fluid studies were positive for C. gattii. The patient was treated with serial lumbar punctures, followed by lumbar drain, as well as amphotericin and flucytosine. The patient had improvement in headache and neurologic symptoms and was discharged to another facility that specializes in management of this disease to undergo further treatment with immunomodulators and steroids. Fungal meningitis is uncommon in the US, particularly among immunocompetent patients. Due to climate change, C. gattii may be a new pathogen to consider. This finding raises important questions to the medical community about the way global climate change affects day to day medical care now, and how it may change in the future.


[WHO classification of Hodgkin's lymphoma and its molecular pathological relevance].
The current WHO classification of Hodgkin's lymphoma (HL) generally distinguishes the relatively rare variant (approximately 5% of all cases of HL) of nodular lymphocyte predominant type from a second group, which comprises classical HL and is separated into four subtypes: lymphocyte rich type, nodular sclerosis type, mixed cellularity type and lymphocyte depleted type. The classical lymphocyte rich subtype is a new entity and based on the typical morphology, can be recognized by definition only by the immunohistochemical characteristics of the Hodgkin and Reed/Sternberg cells (HRS) (CD30+, CD15+, CD20-). Molecular single cell studies are consistent with the dichotomy of HL in nodular lymphocyte predominant and classical types and stress the exceptional position of the former, which shows similarities with non-Hodgkin's lymphomas in several aspects. On the molecular biology level the tumor cells of all kinds of HL turn out to be clonal B cells derived from germinal center cells. However, tumor cells of nodular lymphocyte predominant HL differ from those of classical HL by the pattern of somatic mutations. Considering the inability of HRS cells of classical HL to express a B cell receptor, they should perish under normal conditions. However, they escape from apoptosis by mechanisms so far only partially understood, such as genomic mutations of the l-kappa B gene and the fas receptor gene or probably by down-regulation of B cell markers. In rare cases, HRS cells of HL can also be derived from T cells, as could be demonstrated by single cell analysis. Also, it could be shown by single cell PCR that HL and non-Hodgkin's lymphoma of both B and T cell types can arise from a common precursor. These results suggest that future classifications of HL will not only take into account the morphological and phenotypical profile, but also mechanisms of transformation yet to be discovered.


[Method for recording the parametrical situation in cases of carcinoma of the cervix uteri].
The degree of parametrical infiltration in cases of carcinoma of the cervix uteri could until now be judged by palpation only. Special ultrasound examinations with rectal scanners should be beneficial in future for the recording of the parametrical situation. Although it is not possible to differentiate between inflammatory and cancerous infiltration with this new method, additional punctures under echographic control could be of great value.


Characteristics and outcomes of rhabdomyosarcoma patients with isolated lung metastases from IRS-IV.
To better understand outcomes in children with rhabdomyosarcoma (RMS) and lung-only metastatic disease, the authors reviewed the experience from Intergroup Rhabdomyosarcoma Studies IV Pilot and IV. Patients with lung-only (n = 46) vs other sites of metastatic disease (n = 234) were reviewed using patient charts and the database of Children's Oncology Group (COG). Sixteen percent of patients with RMS and metastatic disease had isolated lung metastases. Thirty-one (67%) had more than 5 metastatic lung lesions. These were bilateral in 34 (74%). Only 6 patients were biopsied at diagnosis. Sixteen children (35%) did not receive any lung radiotherapy. Patients that received lung radiotherapy had fewer lung recurrences ( P = .04), although this has no significant impact on overall survival (OAS, 47% radiotherapy vs 31% no radiotherapy). Compared with patients with other sites of metastatic disease, patients with lung-only metastases have a greater proportion of favorable histology (67% vs 39%, P = .0017), negative nodal involvement (67% vs 32%, P = .0013), and parameningeal primaries (39% vs 12%) and a smaller proportion of extremity primaries (20% vs 33%, P = .0005 for site of primary tumor). Overall survival at 4 years for lung-only metastases was not significantly different from other single-site metastasis (42% vs 34%). Survival was not improved for unilateral disease or fewer than 5 metastatic lesions. Factors associated with diminished OAS include unfavorable histology (P = .0001) and age >10 years (P = .015). Children with RMS and lung-only metastases usually present with extensive bilateral disease that is frequently not biopsied nor given protocol-recommended radiotherapy (XRT). However, outcome is comparable, although slightly better, than patients with other single-site metastasis.


Kinetics of the Trans Effect in Ruthenium Complexes Provide Insight into the Factors That Control Activity and Stability in CO2 Electroreduction.
Comparative kinetic studies of a series of new ruthenium complexes provide a platform for understanding how strong trans effect ligands and redox-active ligands work together to enable rapid electrochemical CO2 reduction at moderate overpotential. After synthesizing isomeric pairs of ruthenium complexes featuring 2'-picolinyl-methyl-benzimidazol-2-ylidene (Mebim-pic) as a strong trans effect ligand and 2,2':6',2″-terpyridine (tpy) as a redox-active ligand, chemical and electrochemical kinetic studies examined how complex geometry and charge affect the individual steps and overall catalysis of CO2 reduction. The relative trans effect of picoline vs the N-heterocyclic carbene (NHC) was quantified through a kinetic analysis of reductively triggered chloride dissociation, revealing that chloride loss is 1000 times faster in the isomer with the NHC trans to chloride. The kinetics of CO dissociation from a site trans to the NHC were examined in a systematic study of isostructural carbonyl complexes across four different overall charges. The rate constants for CO loss span 12 orders of magnitude and are fastest upon two-electron reduction, leading to a hypothesis that redox-active ligands play a key role in promoting reductive CO dissociation during catalysis. Analogous studies of complexes featuring the picoline ligand trans to the carbonyl reveal the importance of the trans effect of the CO ligand itself, with picoline ligand dissociation observed upon reduction. The complexes with NHC trans to the active site proved to be active electrocatalysts capable of selective CO2 electroreduction to CO. In acidic solutions under a N2 atmosphere, on the other hand, H2 evolution proceeds via an intermediate that positions a hydride ligand trans to picoline. The mechanistic insight and quantitative kinetic parameters that arise from these studies help establish general principles for molecular electrocatalyst design.


The role of allergy in the pathogenesis of otitis media with effusion.
To explain an allergic basis for the development of otitis media with effusion (OME), it was suggested that the middle ear mucosa can act as an allergic "shock organ." To evaluate this possibility, 16 juvenile rhesus monkeys were passively sensitized to pollen by intravenous injection of allergic human serum. All ears were then challenged by insufflation of pollen via the nose and eustachian tube (ET), twice daily, for four to five days. Daily tympanometry and otomicroscopy were performed, and on the last day of challenge, tympanocentesis was done to recover effusions. Five animals were killed and the middle ears were processed for histologic study. The results showed that none of the ears developed a middle ear effusion or OME. It is concluded that middle ear challenge with an appropriate pollen antigen in passively sensitized rhesus monkeys does not initiate an inflammatory reaction in the middle ear or induce OME.


Eliciting Preferences for Health Insurance in Iran Using Discrete Choice Experiment Analysis.
The preferences of Iranians concerning the attributes of health insurance benefit packages are not well studied. This study aimed to elicit health insurance preferences among insured people in Iran during 2016. A mixed methods study using a discrete choice experiment (DCE) approach was conducted to elicit health insurance preferences on a total sample of 600 insured Iranians residing in Tehran. The final design of the DCE included 8 health insurance attributes. Data were analyzed using conditional logistic regression models. The final model of this DCE study included 8 attributes, and the findings indicated statistically significant (P<.001) increase in the odds ratio (OR) of choosing health insurance at all levels of cost coverage except for the rehabilitation and para-clinical benefits, where at 70% cost coverage there was insignificant (P=.485) disutility (OR=0.95). With the increase in cost coverage level, the probability of choosing health insurance was significantly (P<.001) the highest for the private hospitals' benefits (OR=2.82) followed by public hospitals' benefits (OR=2.02) and outpatient benefits (OR=1.75), and the premium revealed statistically significant (P<.001) disutility (OR=0.96). Our findings revealed that participants would be willing to choose health insurance plans with higher cost coverage of healthcare services and with lower premiums. However, the demographic characteristics, income, and health status of the insured individuals affected their health insurance preferences. The findings can contribute to the design of better health insurance policies, improve the participation of individuals in health insurance, and increase the insured individuals' utility from the insurance benefits packages.


A mutation screen in patients with Kabuki syndrome.
Kabuki syndrome (KS) is one of the classical, clinically well-known multiple anomalies/mental retardation syndromes, mainly characterized by a very distinctive facial appearance in combination with additional clinical signs such as developmental delay, short stature, persistent fingerpads, and urogenital tract anomalies. In our study, we sequenced all 54 coding exons of the recently identified MLL2 gene in 34 patients with Kabuki syndrome. We identified 18 distinct mutations in 19 patients, 11 of 12 tested de novo. Mutations were located all over the gene and included three nonsense mutations, two splice-site mutations, six small deletions or insertions, and seven missense mutations. We compared frequencies of clinical symptoms in MLL2 mutation carriers versus non-carriers. MLL2 mutation carriers significantly more often presented with short stature and renal anomalies (p = 0.026 and 0.031, respectively), and in addition, MLL2 carriers obviously showed more frequently a typical facial gestalt (17/19) compared with non-carriers (9/15), although this result was not statistically significant (p = 0.1). Mutation-negative patients were subsequently tested for mutations in ten functional candidate genes (e.g. MLL, ASC2, ASH2L, and WDR5), but no convincing causative mutations could be found. Our results indicate that MLL2 is the major gene for Kabuki syndrome with a wide spectrum of de novo mutations and strongly suggest further genetic heterogeneity.


Membrane-traversing mechanism of thyroid hormone transport by monocarboxylate transporter 8.
Monocarboxylate transporter 8 (MCT8) mediates thyroid hormone (TH) transport across the plasma membrane in many cell types. In order to better understand its mechanism, we have generated three new MCT8 homology models based on sugar transporters XylE in the intracellular opened (PDB ID: 4aj4) and the extracellular partly occluded (PDB ID: 4gby) conformations as well as FucP (PDB ID: 3o7q) and GLUT3 (PDB ID: 4zwc) in the fully extracellular opened conformation. T3-docking studies from both sides revealed interactions with His192, His415, Arg445 and Asp498 as previously identified. Selected mutations revealed further transport-sensitive positions mainly at the discontinuous transmembrane helices TMH7 and 10. Lys418 is potentially involved in neutralising the charge of the TH substrate because it can be replaced by charged, but not by uncharged, amino acids. The side chain of Thr503 was hypothesised to stabilise a helix break at TMH10 that undergoes a prominent local shift during the transport cycle. A T503V mutation accordingly affected transport. The aromatic Tyr419, the polar Ser313 and Ser314 as well as the charged Glu422 and Glu423 lining the transport channel have been studied. Based on related sugar transporters, we suggest an alternating access mechanism for MCT8 involving a series of amino acid positions previously and newly identified as critical for transport.


Inhibition of bacteriophage phi chi 174 DNA replication in dnaB mutants of Escherichia coli C.
Bacteriophage phiX174 DNA replication was examined in temperature-sensitive dnaB mutants of Escherichia coli C to determine which stages require the participation of the product of this host gene. The conversion of the infecting phage single-stranded DNA to the double-stranded replicative form (parental RF synthesis) is completely inhibited at the nonpermissive temperature (41 C) in two of the three dnaB mutants tested. The efficiency of phage eclipse and of phage DNA penetration of these mutant host cells at 41 C is the same as that of the parent host strain. The defect is most likely in the synthesis of the complementary strand DNA. The semiconservative replication of the double-stranded replicative form DNA (RF replication) is inhibited in all three host mutants after shifting from 30 to 41 C. Late in infection, the rate of progeny single-stranded phage DNA synthesis increases following shifts from 30 to 41 C. Approximately the same amounts of phage DNA and of infectious phage particles are made following the shift to 41 C as in the control left at 30 C. The simplest interpretation of our data is that the product of the host dnaB gene is required for phiX174 parental RF synthesis and RF replication, but is not directly involved in phage single-stranded DNA synthesis once it has begun. The possible significance of the synthesis of parental RF DNA at 41 C in one of the three mutants is discussed.


[A 26-year-old cyclist with intermittent claudication].
A 26-year-old male amateur cyclist, with no risk factors for vascular disease or previous trauma, presented with left-calf claudication. Physical and additional examination revealed an occlusion of the external iliac artery. During the operation, the cause was found to be an endofibrotic lesion of the external iliac artery, probably due to mechanical trauma as a result of the non-physiological aerodynamic position held on the bicycle during many hours of training. An endarterectomy was performed and the tendon of the psoas-minor muscle was cut because of its strong impression on the psoas-major muscle, which resulted in kinking of the external iliac artery. There followed two episodes of re-occlusion which were treated with a venous interposition graft and a dacron interposition graft, respectively. Thereafter the patient was able to train without pain. Intermittent claudication of the legs in young athletes should not be underestimated; occlusive vascular disease caused by arterial endofibrosis should be considered.


Kallikrein-related peptidases in lung diseases.
Human tissue kallikreins (KLKs) are 15 members of the serine protease family and are present in various healthy human tissues including airway tissues. Multiple studies have revealed their crucial role in the pathophysiology of a number of chronic, infectious and tumour lung diseases. KLK1, 3 and 14 are involved in asthma pathogenesis, and KLK1 could be also associated with the exacerbation of this inflammatory disease caused by rhinovirus. KLK5 was demonstrated as an influenza virus activating protease in humans, and KLK1 and 12 could also be involved in the activation and spread of these viruses. KLKs are associated with lung cancer, with up- or downregulation of expression depending on the KLK, cancer subtype, stage of tumour and also the microenvironment. Functional studies showed that KLK12 is a potent pro-angiogenic factor. Moreover, KLK6 promotes malignant-cell proliferation and KLK13 invasiveness. In contrast, KLK8 and KLK10 reduce proliferation and invasion of malignant cells. Considering the involvement of KLKs in various physiological and pathological processes, KLKs appear to be potential biomarkers and therapeutic targets for lung diseases.