Angelman syndrome in adulthood.
Angelman syndrome (AS) is a neurogenetic disorder. The goal of this study was to investigate the primary health issues affecting adults with AS and to further characterize the natural history and genotype-phenotype correlations. Standardized phone interviews with caregivers for 110 adolescents and adults with AS were conducted. The impact of age, sex, and genotype on specific outcomes in neurology, orthopedics, internal medicine, and psychiatry were investigated. The mean age of individuals with AS was 24 years (range 16-50y). Active seizures were present in 41% of individuals, and 72% had sleep dysfunction. Significant constipation was present in 85%, and 32% were overweight or obese, with obesity disproportionately affecting women. Scoliosis affected 50% with a mean age at diagnosis of 12 years, and 24% of those diagnosed with scoliosis required surgery, an intervention disproportionately affecting men. Sixty-eight percent were able to walk independently, and 13% were able to speak 5 or more words. Self-injurious behavior was exhibited in 52% of individuals. The results of this study indicate that epilepsy severity may assume a bimodal age distribution: seizures are typically most severe in early childhood but may recur in adulthood. While late-adolescent and adult sleep patterns were improved when compared to the degree of sleep dysfunction present during infancy and childhood, the prevalence of poor sleep in adults remained quite high. Primary areas of clinical management identified include the following: seizures, sleep, aspiration risk, GERD, constipation, dental care, vision, obesity, scoliosis, bone density, mobility, communication, behavior, and anxiety.


'The poetry of psychiatry': existential analysis and the politics of psychopathology in Franco's Spain.
This article examines the presence and influence of the work of Swiss psychiatrist Ludwig Binswanger and existential analysis (Daseinsanalyse) in Spanish psychiatry in the central decades of the 20th century. First, and drawing on various printed and archival sources, it reconstructs the important personal and professional ties that Binswanger maintained with numerous Spanish colleagues and describes the notable dissemination of his work in Spain through bibliographical reviews, scientific events, academic reports, university lectures and translations. Next, it reviews the incorporation of the postulates of existential analysis into the discourse of Spanish psychiatrists and assesses their most elaborate and original contributions to the foundations of 'anthropological-existential' psychiatry or the 'existential-analytical' interpretation of certain disorders or clinical conditions. And, finally, it tries to clarify the assessment according to which the (inevitable) instrumentalisation of existential analysis in the context of Franco's Spain first compromised the critical recognition of its true possibilities (and limits) and later contributed to the discrediting of psychopathological research among Spanish psychiatrists.


Hydrophobicity Scaling of Aqueous Interfaces by an Electrostatic Mapping.
An understanding of the hydrophobicity of complex heterogeneous molecular assemblies is crucial to characterize and predict interactions between biomolecules. As such, uncovering the subtleties of assembly processes hinges on an accurate classification of the relevant interfaces involved, and much effort has been spent on developing so-called "hydrophobicity maps." In this work, we introduce a novel electrostatics-based mapping of aqueous interfaces that focuses on the collective, long-wavelength electrostatic response of water to the presence of nearby surfaces. In addition to distinguishing between hydrophobic and hydrophilic regions of heterogeneous surfaces, this electrostatic mapping can also differentiate between hydrophilic regions that polarize nearby waters in opposing directions. We therefore expect this approach to find use in predicting the location of possible water-mediated hydrophilic interactions, in addition to the more commonly emphasized hydrophobic interactions that can also be of significant importance.


Changes in proportion of thiol to disulphide in acid-soluble nuclear proteins of Echinus during the first cell cycle.
1. DNA synthesis in Echinus esculentus eggs kept at 10 degrees C takes place just after fusion, 0.75-1.5h after fertilization, and at telophase at about 2.67-3.33h after fertilization. 2. An increase in the thiol/thiol+disulphide ratio in acid extracts from washed nuclear fractions of the eggs is found at fusion, at early stages of mitosis and at telophase. When DNA is being synthesized, the relative amount of thiol in the extracts increases. 3. There are at least five thiol-containing histones in the acid extract together with a diffusible thiol peptide containing methyl-lysine and 3-methylhistidine and a thiol-containing acidic protein.


Constructed wetlands for environmental pollution control: a review of developments, research and practice in Ireland.
For the purpose of synthesizing a compendium of efforts aimed at environmental pollution control through the use of constructed wetlands systems (CWs) in Ireland, a detailed review of CWs was undertaken. Emphasis was placed on the diverse range of development, practice and researches on CWs technology, placing them in the overall context of the need for low-cost and sustainable wastewater treatment systems. The potential use of CWs in protecting estuarine quality within the current legislative framework is considered, as well as the emerging concept of integrated constructed wetlands (ICWs). In addition, an assessment of the efficiency of CWs in operation in Ireland towards abating environmental pollution was done, and compared with CWs operating in other European countries. The need for sufficient and appropriate data to assist in further development of CWs and modelling studies, and instilling confidence in the public is also highlighted.


Tensions Between Policy and Practice in Finnish Biobank Legislation.
In 2012 a new biobanking law came into effect in Finland which, according to some, is one of the most comprehensive and broad national biobanking legislations in the world to date. The law covers both clinical and research collections and provides institutions with great flexibility in terms of transferring existing collections into a biobank. The law also includes provisions for individuals whose samples are stored in the biobank to request information regarding the studies in which the sample has been used. However, this provision also compels biobanks to provide an account of the significance of the research findings to their individual health. This legal provision has created tension between policy and practice in that most biobanks will struggle to interpret and apply this provision in practice. The relationship between the rights and needs to conduct research using biobank material and individual rights of access is sometimes unclear and fraught with interpretational challenges. The derivation of national interpretations of European Directives and Conventions gives rise to what I call interpretive regulatory dissonance. This is expected to be a challenge for harmonizing European and global biobanking activities in the future.


Balloon-occluded retrograde transvenous obliteration for treatment of gastric varices.
Rupture of gastric varices (GVs) can be fatal. Balloon-occluded retrograde transvenous obliteration (BRTO), as known as retrograde sclerotherapy, has been widely adopted for treatment of GVs because of its effectiveness, ability to cure, and utility in emergency and prophylactic treatment. Simplifying the route of blood flow from GVs to the gastrorenal shunt is important for the successful BRTO. This review outlines BRTO indications and contraindications, describes basic BRTO procedures and modifications, compares BRTO with other GVs treatments, and discusses various combination therapies. Combined BRTO and partial splenic embolization may prevent exacerbation of esophageal varices and shows promise as a treatment option.


A Comparative Analysis of Methods of Endmember Selection for Use in Subpixel Classification: A Convex Hull Approach.
Mixed pixels in aerial and satellite images are common, especially near the boundaries of two or more discrete classes; that is, they tend to occur at the transitional region between two classes. Ideally, to decipher the mixed pixel, a soft classification is performed compared to a hard- or a per-pixel classification. Soft or subpixel classification is carried out where the fractional cover of the LULC contained within a pixel is derived. Endmembers are extracted for three VNIR bands of ASTER data for two image datasets using three approaches, namely, principal component analysis (PCA), pixel purity index (PPI), and convex hull-Graham scan (CHGS). On comparing the DN values of the identified endmembers, it is observed that the CHGS method provides the most appropriate end members than the PCA-derived and PPI-derived end members. This is based on deriving the endmembers from two different image conditions. Convex hull implemented using the Graham scan algorithm delineates the pure pixel and pinpoints the exact number of endmembers. These accurate end members would result in accurate proportions of the land cover for better modeling of the terrain.


Case-control study of congenital defects and parental employment in health care.
Health care workers may be occupationally exposed to known and suspected teratogens including viruses, anesthetic gases, sterilants, mercury, and x-radiation. To assess the risk of congenital defects among offspring of health care workers, we analyzed parental occupational histories for 4,915 case babies with congenital defects, registered during the years 1968-1980 by the Metropolitan Atlanta Congenital Defects Program (MACDP) registry, and for 3,027 control babies born without defects during the same period. Offspring of mothers employed in a nursing occupation during the periconceptional period had a modest excess risk of having at least one congenital defect (relative risk [RR] = 1.42; 95% confidence interval [CI] 1.06-1.88); the offspring were at statistically significant increased risk of having anencephaly or spina bifida (RR = 2.00; 95% CI 1.01-4.30), coarctation of the aorta (RR = 2.06; 95% CI 1.10-3.82), genital system defects (RR = 1.61; 95% CI 1.03-2.53), and urinary system defects (RR = 3.43; 95% CI 1.41-8.34). These associations were not confounded by maternal age, education, or alcohol consumption. Offspring of mothers employed in administrative or clerical jobs in the health care industry also had a modest excess risk of defects (RR = 1.35; 95% CI 0.96-1.90), including a statistically significant excess risk of limb defects. We also found associations between neural tube defects and potential exposure to anesthetic gases and to x-radiation, but each association was based on only three case babies of potentially exposed parents. We found no associations between defects and paternal health care employment, except for a few individual defects, and these were based on small numbers of exposed subjects. Only one of five previous studies reviewed found an increased risk of congenital defects among offspring of nurses, but three of the four negative studies had substantially smaller sample sizes than the present study. Detection bias may be a possible explanation for the apparent excess risk of certain defects among offspring of nurses.


Uncoupling of electrotonic synapses by calcium.
The degree of axo-axonal synaptic coupling between nerve cells in the buccal ganglion of Navanax was investigated in relation to intracellular ionic calcium. Increasing intracellular Ca2+ by injection of Ca2+, injection of Na+, or application of ionophore X537A produced uncoupling after at least 90 min, if metabolic inhibitor was present in the medium. Subsequent removal of the metabolic inhibitor reestablished the coupling in less than 30 min. Injected Sr2+ also mimicked the uncoupling action of Ca2+. The presence of a metabolic inhibitor alone had no effect on the coupling. These results lead to the following conclusions: (i) Uncoupling is due to an increased free Ca2+ concentration at the junctions. (ii) The liberation of endogenous sequestered Ca2+ is not sufficient to produce uncoupling except if an excess Ca2+ had been previously sequestered. The electrical synapses in the buccal ganglion of Navanax thus appear to be affected by Ca2+ in a similar way as gap junctions studied in non-neural tissues.


Serial echocardiographic changes in patients on hemodialysis: an evaluation of guideline implementation.
Few studies have addressed the description of serial changes in left ventricular mass (LVM) and relevant risk factors. All these studies were initiated before the implementation of EBPG or K/DOQI guidelines. The aims of our study were to prospectively describe trends in left ventricular (LV) structure and function, evaluate risk factors for progression of LVM derived from serial echocardiographic measurements starting January 2003, 6 months after full implementation of EBPG in our unit. One hundred seventy-six patients were enrolled at baseline, between 1 January 2003 and 1 October 2004; 33 patients were excluded from analysis due to poor echocardiographic window, 14 patients died and 26 were transplanted or transferred during the follow-up period of minimum 12 months. One hundred and three patients were included in the final analysis (mean age 51 years, mean follow-up 24.1 +/- 14.4 months). Echocardiography was performed at inclusion and at the end of study. Biochemical, blood pressure (BP) and medication data were collected and the means of monthly values were used. At baseline, 86.4% of the patients had left ventricular hypertrophy (LVH) (56.3% concentric hypertrophy, 30.1% eccentric hypertrophy, 6.8% concentric remodeling and only 6.8% normal LV geometry), 25.6% had systolic dy-sfunction and 50.5% had abnormal LV volume index (LVVI). Similar data were recorded at follow-up (82.5%, 44.7%, 37.9%, 7.7%, 9.7%, and 19.5%, respectively). Baseline left ventricular mass index (LVMI) significantly correlated with hemoglobin (Hb) and total protein level. LVMI at follow-up correlated to Hb, SBP, PP, mean level of serum phosphate, calcium x phosphate product and cholesterol. Independent predictors for LVMI (multiple regressions) were anemia and mineral metabolism markers. In our population, 62.1% of the patients had a regression of LVMI, with a mean decrease in LVMI of 12 g/m 2 (1.7 +/- 11.7 g/m 2 /month) over more than 12 months of guideline implementation. Regressors had a significant improvement of anemia, serum phosphate level and calcium x phosphate product (p<0.05). Our study suggests that a holistic interventional approach, targeting various pathogenic mechanisms, as per guidelines, can elicit at least a partial regression in LV structural and functional abnormalities in hemodialysis patients.


Isolation and characterization of a bacteriophage tail-like bacteriocin from a strain of Rhizobium.
Bacterial strain 16-3 spontaneously produces a bacteriocin which inhibits the growth of closely related strain 16-2. Both strains were newly isolated from root nodules of lupines and probably belong to the species Rhizobium lupini. Production of infectious progeny of newly isolated virulent phage 16-2-4 in strain 16-2 is inhibited completely if complexes are bacteriocin-treated during the first half of the latent period. Treatment begun during the second half leads to premature lysis of complexes and inactivates only those progeny phages which were not yet fully matured at the moment of the particle-induced lysis. Examination by electron microscope of the bacteriocin enrichment revealed the presence of particles 123 nm in length which resemble the tails of T-even bacteriophages. Since the particles sediment together with the bactericidal activity in the sucrose gradient and adsorb specifically to bacteriocin-sensitive cells, it is concluded that they are identical with the bactericidal agent. The particles are not found attached to phage heads and cannot self-propagate; they are regarded as incomplete and are named INCO particles. INCO particles consist of a core enveloped by a contractile sheath. One end of the sheath is connected to a baseplate to which six fibers, each 32 nm in length, are attached. These connect the baseplate of an adsorbing particle to the cell surface. Since INCo cores are probably empty, it is concluded that specific adsorption of the particles to the bacterial surface is sufficient to inactive sensitive cells irreversibly.


Radiation resonance emission from steep overcritical plasma profiles illuminated by femtosecond laser pulses.
A radiation resonance effect observed in the reflection spectra from overdense plasmas illuminated by femtosecond laser pulses at normal incidence is reported from particle-in-cell simulations. Harmonic emission at multiple orders of the fundamental is found to exhibit resonance phenomena, with the number of resonances and power emitted depending on the electron plasma density. For relatively low laser intensities the reflected light at the laser frequency shows prominent resonant emission around specific values of the plasma density, mainly at 4, 16, and 36 times critical. For increasing laser intensities, strong harmonic emission around 4 and 16 times critical dominates the reflection spectra. In the case of the third laser harmonic, the emission is found to be resonant about those densities and presents, additionally, a distinctive resonant region around nine times critical. A simple radiation model for the power of the third harmonic was proposed confirming a resonant effect dependent on the electron plasma density. For higher harmonic numbers, weak radiation resonances persist in the emission spectra, with their number increasing with order. The resonance effect reported in this paper is found to occur at densities that approximately satisfy n(e)/n(c)=4n(2), where n(e) and n(c) are the plasma and the critical density, respectively, and n is an integer. For the third harmonic, the second resonance corresponds to n=1.5.


Biochanin A inhibits endothelial cell functions and proangiogenic pathways: implications in glioma therapy.
Malignant gliomas, such as glioblastoma multiforme, are highly vascularized tumors of the central nervous system. A rich network of angiogenic vessels supporting glioma growth is an important therapeutic target in glioma therapy. In the past few years, small molecules have gained interest as multitargeting therapies for cancer. Biochanin A is a small, natural dietary isoflavone known for its anticancer potential. Previously, we have found that biochanin A inhibits invasion in human glioblastoma cells. In this study, we elucidated the antiangiogenic mechanisms of biochanin A using rat brain tumor (C6) and murine brain endothelial (bEnd.3) cells and an ex-vivo chick chorioallantoic membrane model. Biochanin A inhibited endothelial cell functions such as cell viability, migration, and invasion, as analyzed using MTT, scratch wound, and gelatin zymography assays. Activation of proangiogenic proteins (ERK/AKT/mTOR) was inhibited. Biochanin A also inhibited chemical hypoxia-inducible factor-1α and vascular endothelial growth factor in C6 cells. Results of chick chorioallantoic membrane assay showed that biochanin A inhibited blood vessel formation ex vivo. As these results suggest that biochanin A directly targets different facets of angiogenesis in vitro and ex vivo, this study provides a rationale for future preclinical evaluation of its efficacy against angiogenic gliomas.


Usefulness of clinical chemistry measurements in classifying patients with breast cancer.
Laboratory measurements can be used to detect, classify, and monitor patients with breast cancer. This review covers in detail the clinical usefulness of carcino embryonic antigen, tissue polypeptide antigen, various glycoproteins, pregnancy-associated proteins, casein, lactalbumin, beta-2-microglobulin, ferritin, immunoglobins, acute phase proteins, placental-like alkaline phosphatase, liver-associated enzymes, glycosyltransferases, human chorionic gonadotropin, calcitonin, polyamines, and collagen breakdown products, in relationship to their diagnostic utility in patients suspected of having or known to have breast cancer. In addition, these authors assess the merits of various multivariate techniques using a number of clinical chemistry quantities in the same regard. Finally, the relative contribution of biochemical tests vs. the information values gained from "surgical pathology" data (number of positive nodes, number of negative nodes, and degree of anaplasia) is discussed.


Purification and biophysical characterization of hepatitis B antigen.
Hepatitis B antigen (HB Ag) has been purified from serum by pepsin digestion and equilibrium sedimentation. The sedimentation coefficient, density, particle size, and electrophoretic mobility have been determined before and after purification and found to be unaltered. In addition, the diffusion constant and molecular weight of purified HB Ag have been documented. Because the biophysical characteristics found are not consistant with those of any known virus, it is believed that native HB Ag is made up almost entirely of incomplete virus.


Chlamydial and gonorrheal neglected sexually transmitted diseases among Pacific Islanders of the Western Pacific Region-A narrative review and call to action.
The Pacific Island countries of the Western Pacific Region have some of the highest rates of sexually transmitted Chlamydia trachomatis and Neisseria gonorrhoeae infections in the world. Despite this, there are few research studies that include Pacific Islanders. We conducted a narrative review of original research and surveys, including World Health Organization and Pacific Community reports, to determine the prevalence, management, and treatment of C. trachomatis and N. gonorrhoeae compared to HIV and syphilis from 1980 to 2022. Available epidemiologic data on C. trachomatis and N. gonorrhoeae indicated an extremely high prevalence-approximately 30% and 13%, respectively-among Pacific Islanders during this timeframe. These neglected sexually transmitted infections represent a significant burden and health disparity. Robust epidemiologic research is needed to identify modifiable risk factors for designing interventions and control strategies. Appropriate policies along with regional and international advocacy and aid are required to improve reproductive health among these vulnerable, understudied populations to avert preventable infections and sequelae.


Efficacy and Toxicity of Concurrent Chemoradiation in Inoperable Oral Carcinoma in Pakistani Population.
To evaluate the efficacy of concurrent chemoradiation in patients with locally advanced inoperable squamous cell carcinoma of oral cavity in terms of local control and toxicity. Case series. Institute of Nuclear Medicine and Oncology (INMOL), Lahore, from January 2008 to December 2013. Sixty-nine patients with locally advanced inoperable oral cavity cancer, registered in INMOL hospital from January 2008 to December 2013 who fulfilled a pre-defined eligibility criteria, were enrolled in the study. Concurrent chemoradiation protocol consisted of conventional fractionation delivering 70 Gy with weekly Cisplatin (50 mg/m2) during the course of radiation. Tumor response was calculated by RECISTcriteria version 1.1 along with the median overall survival and disease-free survival. Acute treatment related toxicities were graded as (G). Thirty-six (52.17%) patients showed complete response; while 19 (27.54%), 8 (11.59%) and 6 (8.7%) were observed with partial response, stable and progressive disease, respectively. Treatment response was significant (p<0.001) in terms of responders vs. non responders to treatment. Median overall survival was 18.00 months; whereas, median disease-free survival remained 14.00 months. Main toxicities included mucositis (G3 and G4, 71%), xerostomia (G2 and G3, 82.5%), vomiting (G3 and G4, 51%), myelosuppression (G3 and G4, 26.2%), dermatitis (G3 and G4, 49.2%), and fatigue (G3 and G4, 57.9%). Platinum based CCR Tremained effective for inoperable oral cancer patients.


Complete genome sequence of Butyricimonas faecihominis JCM 18676T.
We report a complete genome sequence of Butyricimonas faecihominis JCM 18676T, generated by nanopore sequencing. The genome consists of a single circular chromosome of 4,851,806 bp, with a G + C content of 42.9%, and was predicted to contain 15 rRNA and 61 tRNA genes and encode for 3,946 proteins.


[Emphasis on standardization of minimally invasive surgery for gastric cancer].
Different proficiency in minimally invasive surgery has great impact on efficacy of minimally invasive surgery. Therefore, great importance should be attached to the standardization of minimally invasive surgery. Accurate preoperative staging plays a prerequisite role which emphasizes multiple diagnostic strategies. Precise intraoperative procedure plays an essential role which should be performed according to the evidence-based expert consensus and guideline. Standardized postoperative management plays a crucial role which includes ex vivo lymph node dissection, registry of complication and follow-up and establishment of medical database. Establishing training system and improving quality standard system will promote standardized utilization of minimally invasive surgery clinically.


Rapid prenatal diagnosis of glycogen-storage disease type II by electron microscopy of uncultured amniotic-fluid cells.
Glycogen-storage disease Type IIa is a fatal, genetically determined disease of infancy or early childhood that is characterized by deficient activity of acid alpha-glucosidase and by the presence of intracellular vacuoles full of glycogen, which are found in most tissues, including skin and liver. On electron microscopy these specific vacuoles are tightly packed accumulations of glycogen particles surrounded by a single membrane. We did electron-microscopical examinations on uncultured amniotic-fluid cells from 26 women whose fetuses were at risk for glycogen-storage disease Type IIa and from 8 normal control pregnant women. We found specific vacuoles in cells from 6 of the 26 high-risk patients. At delivery, glycogen-storage disease Type IIa was present in the infants of these 6 women and absent in those of the other 20 according to results of clinical, biochemical, and electron-microscopical studies of gestational products. After amniocentesis at 15 to 18 weeks of gestation, the prenatal diagnosis made by electron microscopy of uncultured amniotic-fluid cells was available in three to six days, whereas it took from three to six weeks to make the diagnosis by enzymatic analysis of the cultured amniotic-fluid cells. We conclude that the electron-microscopical prenatal diagnosis of glycogen-storage disease Type IIa is rapid, safe, and reliable. It should facilitate earlier diagnosis and thereby help to preserve parental options.


Trichothecene production by Trichoderma brevicompactum.
Trichoderma brevicompactum, T. viride, T. harzianum, T. atroviride, T. longibrachiatum, T. erinaceum, T. citrinoviride, and Hypocrea lutea were screened for production of trichothecenes after growth on one or several solid and liquid media. Trichothecenes were detected by liquid chromatography combined with online UV/vis spectroscopy and electrospray high-resolution mass spectrometry. T. brevicompactum produced trichodermin and/or harzianum A on all media investigated, with liquid media yielding the largest amounts. Detection of octa-2Z,4E,6E-trienedioic acid in the harzianum-A-producing strains indicated that harzianum A was synthesized directly by esterification of trichodermol with octa-2Z,4E,6E-trienedioic acid. Both the T. viride strain from which trichodermin was originally isolated and the T. harzianum strain from which harzianum A was originally isolated were shown to belong to T. brevicompactum based on four independent criteria: metabolite profiles, micromorphology, macromorphology on yeast extract sucrose agar and potato dextrose agar, and DNA sequences of the ITS1/ITS2 regions of the nuclear ribosomal DNA.


A practical synthesis of alpha-D-Manp-(1-->3)-alpha-D-Manp-(1-->2)-[alpha-D-Glcp-(1-->3)]-alpha-D-Manp-(1-->2)-alpha-D-Manp-(1-->2)-alpha-D-Manp, an O-specific heterohexasaccharide fragment of Citrobacter braakii O7a, 3b, 1c.
An O-specific heterohexasaccharide fragment of Citrobacter braakii O7a, 3b, 1c, alpha-D-Manp-(1-->3)-alpha-D-Manp-(1-->2)-[alpha-D-Glcp-(1-->3)]-alpha-D-Manp-(1-->2)-alpha-D-Manp-(1-->2)-alpha-D-Manp was synthesized as its methyl glycoside. Acetylation of allyl 4,6-O-benzylidene-alpha-D-mannopyranoside, followed by debenzylidenization and benzoylation gave allyl 2,3-di-O-acetyl-4,6-di-O-benzoyl-alpha-D-mannopyranoside (3), and subsequent deacetylation of 3 with CH(3)COCl-MeOH gave the monosaccharide acceptor 4. Condensation of isopropyl 2,3,4,6-tetra-O-benzyl-1-thio-beta-D-glucopyranoside (6) with 4 selectively afforded the alpha-(1-->3)-linked disaccharide 7. Condensation of 7 with the (1-->3)-linked disaccharide donor 9, followed by deallylation and trichloroacetimidation, afforded the tetrasaccharide donor 12. Coupling of 12 with disaccharide acceptor 13, followed by debenzylation and deacylation, furnished the target heterohexasaccharide 16.


A comprehensive, non-invasive visualization of primordial germ cell development in mice by the Prdm1-mVenus and Dppa3-ECFP double transgenic reporter.
The ability to monitor the development of a given cell lineage in a non-invasive manner by fluorescent markers both in vivo and in vitro provides a great advantage for the analysis of the lineage of interest. To date, a number of transgenic or knock-in mouse strains, in which developing germ cells are marked with fluorescent reporters, have been generated. We here describe a novel double transgenic reporter mouse strain that expresses membrane-targeted Venus (mVenus), a brighter variant of yellow fluorescent protein (YFP), under the control of Prdm1 (Blimp1) regulatory elements and enhanced cyan fluorescent protein (ECFP) under the control of Dppa3 (Stella/Pgc7). The double transgenic strain unambiguously marked Prdm1 expression in the lineage-restricted precursors of primordial germ cells (PGCs) in the proximal epiblast at embryonic day (E) 6.25 and specifically illuminated Prdm1- and Dppa3-positive migrating PGCs after E8.5. The double transgenic reporter also precisely recapitulated dynamic embryonic expression of Prdm1 outside the germ cell lineage. Moreover, we derived ES cells that bore both transgenes. These cells made a robust contribution both to the germ and somatic cell lineages in chimeras with accurate Prdm1-mVenus and Dppa3-ECFP expression. The transgenic strain and the ES cells will serve as valuable experimental materials not only for analyzing the origin and properties of the germ cell lineage in vivo, but also for establishing a culture system to efficiently induce proper germ cells with temporally coordinated Prdm1 and Dppa3 expression in vitro.


Seroprevalence of anti-SARS-CoV-2 antibodies among school and daycare children and personnel: protocol for a cohort study in Montreal, Canada.
Further evidence is needed to understand the contribution of schools and daycares for the spread of COVID-19 in the context of diverse transmission dynamics and continually evolving public health interventions. The Enfants et COVID-19: Étude de séroprévalence (EnCORE) study will estimate the seroprevalence and seroconversion of SARS-CoV-2 among school and daycare children and personnel. In addition, the study will examine associations between seroprevalence and sociodemographic characteristics and reported COVID-19 symptoms and tests, and investigates changes in health, lifestyle and well-being outcomes. This study includes children and personnel from 62 schools and daycares in four neighbourhoods in Montreal, Canada. All children aged 2-17 years attending one of the participating schools or daycares and their parents are invited to participate, as well as a sample of personnel members. Participants respond to brief questionnaires and provide blood samples, collected via dried blood spot, at baseline (October 2020-March 2021) and follow-up (May-June 2021). Questionnaires include sociodemographic and household characteristics, reported COVID-19 symptoms and tests, potential COVID-19 risk factors and prevention efforts and health and lifestyle information. Logistic regression using generalised estimating equations will be used to estimate seroprevalence and seroconversion, accounting for school-level clustering. This study was approved by the research ethics boards of the Université de Montréal (CERSES) and the Centre Hospitalier Universitaire Sainte-Justine. Results will contribute to our knowledge about SARS-CoV-2 transmission in schools and daycares and will be made available to study participants and their families, school and public health decision-makers and the research community.


Multi-Feature Classification of Multi-Sensor Satellite Imagery Based on Dual-Polarimetric Sentinel-1A, Landsat-8 OLI, and Hyperion Images for Urban Land-Cover Classification.
This paper focuses on evaluating the ability and contribution of using backscatter intensity, texture, coherence, and color features extracted from Sentinel-1A data for urban land cover classification and comparing different multi-sensor land cover mapping methods to improve classification accuracy. Both Landsat-8 OLI and Hyperion images were also acquired, in combination with Sentinel-1A data, to explore the potential of different multi-sensor urban land cover mapping methods to improve classification accuracy. The classification was performed using a random forest (RF) method. The results showed that the optimal window size of the combination of all texture features was 9 × 9, and the optimal window size was different for each individual texture feature. For the four different feature types, the texture features contributed the most to the classification, followed by the coherence and backscatter intensity features; and the color features had the least impact on the urban land cover classification. Satisfactory classification results can be obtained using only the combination of texture and coherence features, with an overall accuracy up to 91.55% and a kappa coefficient up to 0.8935, respectively. Among all combinations of Sentinel-1A-derived features, the combination of the four features had the best classification result. Multi-sensor urban land cover mapping obtained higher classification accuracy. The combination of Sentinel-1A and Hyperion data achieved higher classification accuracy compared to the combination of Sentinel-1A and Landsat-8 OLI images, with an overall accuracy of up to 99.12% and a kappa coefficient up to 0.9889. When Sentinel-1A data was added to Hyperion images, the overall accuracy and kappa coefficient were increased by 4.01% and 0.0519, respectively.


Kinetic investigation of the reactions of S-4-nitrophenyl 4-substituted thiobenzoates with secondary alicyclic amines in aqueous ethanol.
The reactions of S-4-nitrophenyl 4-X-substituted thiobenzoates (X = H, Cl, and NO(2): 1, 2, and 3, respectively) with a series of secondary alicyclic amines (SAA) were subjected to a kinetic investigation in 44 wt % ethanol-water, at 25.0 degrees C and an ionic strength of 0.2 M (KCl). The reactions were followed spectrophotometrically by monitoring the release of 4-nitrobenzenethiolate anion at 420-425 nm. Under excess amine, pseudo-first-order rate constants (k(obsd)) are obtained for all reactions. The plots of k(obsd) vs [SAA] at constant pH are linear with the slope (k(N)) independent of pH. The statistically corrected Brønsted-type plots (log k(N)/q vs pK(a) + log p/q) for the reactions of 1 and 2 are nonlinear with slopes at high pK(a), beta(1) = 0.27 and 0.10, respectively, and slopes at low pK(a), beta(2) = 0.86 and 0.84, respectively. The Brønsted curvature is centered at pK(a) (pK(a)(0)) 10.0 and 10.4, respectively. The reactions of SAA with 3 exhibit a linear Brønsted-type plot of slope 0.81. These results are consistent with a stepwise mechanism, through a zwitterionic tetrahedral intermediate (T(+/-)). For the reactions of 1 and 2, there is a change in rate-determining step with amine basicity, from T(+/-) breakdown to products at low pK(a), to T(+/-) formation at high pK(a). For the reactions of 3, breakdown to products of T(+/-) is rate limiting for all the SAA series (pK(a)(0) > 11). The increasing pK(a)(0) value as the substituent in the acyl group becomes more electron withdrawing is attributed to an increasing nucleofugality of SAA from T(+/-). The greater pK(a)(0) value for the reactions of SAA with 1, relative to that found in the pyridinolysis of 2,4-dinitrophenyl benzoate (pK(a)(0) = 9.5), is explained by the greater nucleofugality from T(+/-) of the former amines, compared to isobasic pyridines, and the greater leaving ability from T(+/-) of 2,4-dinitrophenoxide relative to 4-nitrobenzenethiolate.


The mobility of 137Cs and 90Sr in agricultural soils in the Ukraine, Belarus, and Russia, 1991.
Results are presented from studies concerning the behavior of the Chernobyl-derived radionuclides 137Cs and 90Sr in soil-plant agricultural systems in the Ukraine, Belarus, and Russia during 1991. The sites, representing ploughed and natural pastures, were located at varying distances between 50 and 650 km and varying directions from the Chernobyl reactor site. The 137Cs activity concentrations in the upper 0-5 cm soil layer ranged from 25-1,000 kBq m-2 and were higher in natural pastures as compared to ploughed pastures. For 90Sr, activity levels ranged from 1.4-40 kBq m-2, and the highest 90Sr deposition was observed in the Gomel Region, Belarus. The highest 90Sr: 137Cs ratio was also observed in the Gomel soils, i.e., 15% as compared to between 0.72 and 7.4% in the other soils. The mobility of radionuclides was studied by means of sequential extraction. For all soils, between 60 and 95% of the 137Cs was found to be strongly bound to soil components. In the Russian and Ukrainian soils, between 40 and 98% of the 90Sr was found in the easily extractable fractions, and the distribution of 137Cs and 90Sr followed that of the naturally occurring stable isotopes of cesium and strontium. However, in the Gomel soils, between 20 and 50% of the 90Sr was easily extractable and the distribution of 90Sr within the extraction fractions did not follow that observed for stable strontium. These results are thought to reflect the association of 90Sr with fuel particles deposited in the Gomel Region. The mobility of 90Sr is expected to increase with time (as the particles weather) in these soils.


The fine-tuned crosstalk between lysine acetylation and the circadian rhythm.
Circadian rhythm is a roughly 24-h wake and sleep cycle that almost all of the organisms on the earth follow when they execute their biological functions and physiological activities. The circadian clock is mainly regulated by the transcription-translation feedback loop (TTFL), consisting of the core clock proteins, including BMAL1, CLOCK, PERs, CRYs, and a series of accessory factors. The circadian clock and the downstream gene expression are not only controlled at the transcriptional and translational levels but also precisely regulated at the post-translational modification level. Recently, it has been discovered that CLOCK exhibits lysine acetyltransferase activities and could acetylate protein substrates. Core clock proteins are also acetylated, thereby altering their biological functions in the regulation of the expression of downstream genes. Studies have revealed that many protein acetylation events exhibit oscillation behavior. However, the biological function of acetylation on circadian rhythm has only begun to explore. This review will briefly introduce the acetylation and deacetylation of the core clock proteins and summarize the proteins whose acetylation is regulated by CLOCK and circadian rhythm. Then, we will also discuss the crosstalk between lysine acetylation and the circadian clock or other post-translational modifications. Finally, we will briefly describe the possible future perspectives in the field.


Biomarkers of intrahepatic cholangiocarcinoma: diagnosis and response to therapy.
Intrahepatic cholangiocarcinoma (ICC) is the second most common primary liver cancer behind hepatocellular carcinoma (HCC) and carries a dismal prognosis. Improved genetic analysis has paved the way for a better understanding of the distinct somatic genomic landscapes of ICC. The use of next generation sequencing has paved the way for more personalized medicine through identifying unique mutations which may prove to be therapeutic targets. The ability to identify biomarkers specific to ICC will assist in establishing a diagnosis, monitoring response to therapy, as well as assist in identifying novel therapies and personalized medicine. Herein, we discuss potential biomarkers for ICC and how these markers can assist in diagnosis, monitor response to therapy, and potentially identify novel interventions for the treatment of ICC.


Draining Fluids through a Peritoneal Catheter in Newborns after Cardiac Surgery Helps to Control Fluid Balance.
Dialysis can be used in severe cases, but may not be well tolerated. In such patients, peritoneal drainage could be an alternative option for fluid removal. We report the case of a newborn with a truncus arteriosus who developed postoperatively a complicated clinical course with right ventricular dysfunction, prerenal condition as well as fluid overload despite diuretic therapy. Dialysis was indicated for fluid removal. Peritoneal dialysis was started using a surgically placed Tenckhoff catheter and stopped due to inefficacy and leaks and no other modalities of dialysis were used. However, the catheter was left in place over a period of two months for fluid drainage and removed because of unexplained fever. In order to determine the effect of peritoneal drainage, we selected a period of one week before and one week after the removal of the drain to compare daily clinical data, urine electrolytes and renal function and found a positive effect on fluid balance control. We conclude that the fluid removal by continuous peritoneal drainage is a simple and safe alternative that can be used to control fluid balance in infants after cardiac surgery.


Visualization of jet development in laser-induced plasmas.
Laser-induced plasmas in gases are known to generate gaseous jets in the postplasma gas plume. The gaseous jet typically develops toward the laser source, and the experiments presented here show, for the first time to our knowledge, that, under certain conditions, these jets can develop in the opposite direction or may not form at all. The data suggest that this is related to the ratio between the energy absorbed in the plasma and the threshold breakdown energy, effectively leading to multiple plasma initiation sites in the focal waist.


Treatment of infertility in women with pituitary tumors.
Due to the pituitary's critical position, a pituitary tumor may disrupt gonadal function, either by its expanding size or the inappropriate secretion of hormones. Menstrual cycles may be disrupted even without frank hypogonadism, particularly in the case of hormone-secreting adenomas. Despite optimal medical and surgical management of pituitary tumors, ovulation-induction therapy with gonadotropins is often required to restore fertility in these women. This article will provide an overview of the therapeutic options available for women with infertility resulting from pituitary tumors. Treatment strategies including dopamine agonists, gonadotropins and the role of assisted reproductive technologies will be discussed. Unique pregnancy considerations in the female patient with hypopituitarism will also be addressed.


Candidemia in patients with ventricular assist devices.
During the period 1998-2004, candidemia developed in 7 of 117 ventricular assist device recipients at our hospital, and the associated mortality rate was 71%. Five cases of candidemia were due to Candida parapsilosis, and 2 were due to Candida albicans. Three of the 7 patients with ventricular assist device-associated Candida bloodstream infections were cured, and the device was retained in 2 of the 3 patients.


Involvement of the 5'-leader sequence in coupling the stability of a human H3 histone mRNA with DNA replication.
Two lines of evidence derived from fusion gene constructs indicate that sequences residing in the 5'-nontranslated region of a cell cycle-dependent human H3 histone mRNA are involved in the selective destabilization that occurs when DNA synthesis is terminated. The experimental approach was to construct chimeric genes in which fragments of the mRNA coding regions of the H3 histone gene were fused with fragments of genes not expressed in a cell cycle-dependent manner. After transfection in HeLa S3 cells with the recombinant plasmids, levels of fusion mRNAs were determined by S1 nuclease analysis prior to and following DNA synthesis inhibition. When the first 20 nucleotides of an H3 histone mRNA leader were replaced with 89 nucleotides of the leader from a Drosophila heat-shock (hsp70) mRNA, the fusion transcript remained stable during inhibition of DNA synthesis, in contrast to the rapid destabilization of the endogenous histone mRNA in these cells. In a reciprocal experiment, a histone-globin fusion gene was constructed that produced a transcript with the initial 20 nucleotides of the H3 histone mRNA substituted for the human beta-globin mRNA leader. In HeLa cells treated with inhibitors of DNA synthesis and/or protein synthesis, cellular levels of this histone-globin fusion mRNA appeared to be regulated in a manner similar to endogenous histone mRNA levels. These results suggest that the first 20 nucleotides of the leader are sufficient to couple histone mRNA stability with DNA replication.


The inclusion of downy mildews in a multi-locus-dataset and its reanalysis reveals a high degree of paraphyly in Phytophthora.
Pathogens belonging to the Oomycota, a group of heterokont, fungal-like organisms, are amongst the most notorious pathogens in agriculture. In particular, the obligate biotrophic downy mildews and the hemibiotrophic members of the genus Phytophthora are responsible for a huge variety of destructive diseases, including sudden oak death caused by P. ramorum, potato late blight caused by P. infestans, cucurbit downy mildew caused by Pseudoperonospora cubensis, and grape downy mildew caused by Plasmopara viticola. About 800 species of downy mildews and roughly 100 species of Phytophthora are currently accepted, and recent studies have revealed that these groups are closely related. However, the degree to which Phytophthora is paraphyletic and where exactly the downy mildews insert into this genus in relation to other clades could not be inferred with certainty to date. Here we present a molecular phylogeny encompassing all clades of Phytophthora as represented in a multi-locus dataset and two representatives of the monophyletic downy mildews from divergent genera. Our results demonstrate that Phytophthora is at least six times paraphyletic with respect to the downy mildews. The downy mildew representatives are consistently nested within clade 4 (contains Phytophthora palmivora), which is placed sister to clade 1 (contains Phytophthora infestans). This finding would either necessitate placing all downy mildews and Phytopthora species in a single genus, either under the oldest generic name Peronospora or by conservation the later name Phytophthora, or the description of at least six new genera within Phytophthora. The complications of both options are discussed, and it is concluded that the latter is preferable, as it warrants fewer name changes and is more practical.


Identification of peptides in the adenine ring binding domain of glutamate and lactate dehydrogenase using 2-azido-NAD+.
Previously, the synthesis and validation of [32P]2N3NAD+ as an active site directed photoaffinity probe for glutamate dehydrogenase (GDH) was reported (8). This report shows that 2N3NAD+ is also an effective probe for the NAD+ binding site of lactate dehydrogenase (LDH). With the appropriate photolabeling procedures and immobilized boronate column chromatography the active site peptides of GDH and LDH involved in the adenine base binding domain have been isolated and sequenced. With both GDH and LDH a single photolabeled peptide, which contained the majority of the photoinserted radiolabel, was isolated. Additionally, these peptides had UV spectra that were markedly different from the nonphotolabeled peptides. The modified peptide from GDH corresponded to Cys270 through Lys289. Both sequencing and compositional analysis identified Glu275 as the site of photoinsertion. Sequencing of this peptide aborted at Glu275 after five rounds of analysis, indicating that insertion was blocking further progress. Compositional analysis showed that the entire sequence from residues 270 to 289 was present except that the single Glu residue was missing. This is interpreted as indicating that the photoinsertion is into the polypeptide backbone at the Glu site. The peptide isolated from LDH corresponded to Asp82 through Arg90. Sequencing of this peptide could be completed throughout with only the round at Tyr83 giving no identifiable residue. Compositional analysis of this peptide was in agreement with the peptide from Asp82 to Arg90 with the exception that the single Tyr residue was missing. This indicates that the photoinsertion is into the tyrosine side chain. This data was found to be in agreement with X-ray crystallographic results identifying the NAD(+)-binding domains.


Immune suppression in healthy carriers of adult T-cell leukemia retrovirus (HTLV-I): impairment of T-cell control of Epstein-Barr virus-infected B-cells.
We have examined the activities of Epstein-Barr virus (EBV)-specific memory T-cells of carriers of adult T-cell leukemia (ATL) associated retrovirus (HTLV-I) and non-carriers in an ATL-endemic area, all of whom were EBV-seropositive, by assay of EBV-induced B-cell focus regression. The result showed that immune suppression, as represented by lowering of the regression, was present in 18 (29%) out of 63 healthy HTLV-I carriers, in contrast to none of 63 matched control persons (healthy non-carriers). This finding suggests that a suppression of cell-mediated immunity is induced by a persistent infection with ATL retrovirus.


Role of toll-like receptor 10 gene polymorphism and gastric mucosal pattern in patients with chronic gastritis.
Helicobacter pylori stimulates the host's toll-like receptors (TLRs). Single-nucleotide polymorphism (SNP) of TLRs is related to the manipulation of regulatory cytokines and also implicated in the varied outcomes of the inflammatory response, including the development of precancerous lesions of gastric mucosa and disease progression. We hypothesized that TLR10 rs10004195 polymorphism is associated with gastric mucosal patterns. TLR10 rs10004195 polymorphisms were identified in a total of 400 gastritis patients using the TagMan SNP genotyping assay. Gastric mucosal patterns were classified by Conventional Narrow Band Imaging gastroscopy (C-NBI gastroscopy). Logistic regression was used to analyze the association. The gastritis patients was Type 1, 37.5% of Thai patients. The T/T homozygous genotype was exhibited by the highest percentage (46.5%) of patients, and the A/A homozygous and A/T heterozygous genotypes were exhibited by 20.25% and 33.25%, respectively, of patients. TLR10 rs10004195 was significantly associated with gastric mucosal patterns. After adjusting for confounding factors, patients with the A/A homozygous genotype showed a significantly increased risk of severe inflammation (OR=1.35, 95% CI=0.97-2.13, p=0.028). Patients with the A/T heterozygous and T/T homozygous genotypes showed a significantly increased risk of mild inflammation (OR=1.24, 95% CI=0.78-2.07, p=0.042 and OR=1.78, 95% CI=0.51-3.35, p=0.001, respectively). Our results indicate that the presence of TLR10 rs10004195, A/T heterozygous, and T/T homozygous genotypes is associated with type 1, 2, and 3 whereas that of the A/A homozygous genotype is associated with type 4 and 5 of gastric mucosal patterns. This suggests that the A/A homozygous genotype contributes to severe inflammation in H. pylori-associated gastritis in Thai patients.


Update on inflation of journal prices: Brandon/Hill list journals and the scientific, technical, and medical publishing market.
The original study of journal prices, using the "Brandon/ Hill Selected List of Books and Journals for the Small Medical Library," was first published in 1980 and periodically updated. This research continues to measure price increases for these titles for the periods 1996 to 1999 and 1999 to 2002. The 111 journal titles that have appeared in each published list from 1967 to 2001 were included in the study. Institutional subscription price data were gathered for each journal for the years 1996, 1999, and 2002 and were compared to the Consumer Price Index (CPI) for the same years. The average journal price continues to rise significantly and is independent of the CPI. The study found that prices have jumped 51.9% from 1996 to 1999 and 32% from 1999 to 2002, which is consistent with nearly every recent journal price study. The unprecedented rise in journal prices negatively affects the purchasing power of medical libraries. This paper examines the economic and technological pressures on the science, technology, and medical journals market that contribute to high prices and identifies a number of initiatives in the biological and health sciences that utilize alternative models for disseminating scientific research.


Lytic induction of Kaposi's sarcoma-associated herpesvirus in primary effusion lymphoma cells with natural products identified by a cell-based fluorescence moderate-throughput screening.
Kaposi's sarcoma-associated herpesvirus (KSHV) has been linked to Kaposi's sarcoma primary effusion lymphoma (PEL), and multicentric Castleman's disease. Intentional lytic induction of gammaherpesviruses in the presence of antiviral drugs is thought to be an effective treatment option for gammaherpesvirus-related tumors. In this study, we used a cell-based fluorescence bioassay system in which a KSHV-infected PEL cell line was stably transfected with a potent viral-promoter-driven reporter gene to identify effective non-toxic reagents capable of inducing latent KSHV. Among 400 plant extracts screened, three extracts increased reporter gene expression in a dose-dependent manner. Furthermore, the three extracts activated the RTA promoter and induced expression of lytic genes in the endogenous viral genomes of KSHV-infected tumor cells. Together, our results demonstrate the effectiveness of a moderate-throughput screening system to identify natural products capable of inducing KSHV reactivation, thereby facilitating the development of novel therapeutic agents for KSHV-associated malignancies.


Quantitative backscattered electron imaging of field emission scanning electron microscopy for discrimination of nano-scale elements with nm-order spatial resolution.
Discrimination of thin film elements by backscattered electron (BSE) imaging of field emission scanning electron microscope was examined. Incident electron acceleration voltage dependence on thin films' BSE intensities in five elements (Au, Ag, Ge, Cu and Fe) on a silicon substrate was experimentally measured from 3 to 30 kV. Normalization of BSE intensities using the difference between maximum and minimum brightness was proposed and allowed reproducible comparison among the elements. Measured intensities, which have correlation with electron backscattering coefficient against atomic number, indicated the existence of adequate acceleration voltage for improvement of resolution to discriminate different elements, showing the possibility of discriminating at least these six elements simultaneously by BSE imaging with nanometer-scale spatial resolution.


Morbidity after preoperative radiotherapy and cystectomy in patients with bladder cancer.
The therapy-related morbidity was evaluated in 121 patients with muscle-invasive or recurrent superficial bladder cancer treated with radiotherapy and cystectomy. In 103 patients cystectomy succeeded 39-52 Gray (Gy) preoperative irradiation and in 18 patients cystectomy was done as a salvage procedure after previous full-dose radiotherapy. The overall frequency of complications was high; 71% of the patients treated with preoperative and 78% treated with full-dose radiotherapy had clinically relevant complications related to radiotherapy or surgery or both. The rate of intestinal complications was 39% for preoperative and 67% for full-dose radiotherapy. The overall mortality rate in intestinal complications was 3.3%. This study shows that the combination of radiotherapy and radical surgery in patients with bladder cancer is associated with a high rate of intestinal complications. The complications are significantly related to the irradiation dose and are long lasting and even life threatening.


Risk factors for rampant caries in children from southwestern Nigeria.
The cross sectional study tried to identify some risk factors for rampant caries in a hospital based population of Nigerian children. 205 consecutive eligible children seen in the paediatric units of two hospitals in Nigeria were recruited into the study. All enrolled children were healthy with no chronic medical condition. Consent for study participation was received from their accompanying parent. The accompanying parent of each child was questioned on the duration of breast and bottle feeding, and form of breast feeding and the frequency of daily intake of sugary diet. The associations between sex, age of the child, frequency of daily consumption of sugar, birth rank, duration of breast feeding, form of breast feeding and duration of bottle feeding and rampant caries were established. No association was found between the occurrence of rampant caries and sex, birth rank, duration of breast feeding, form of breast feeding and duration of bottle feeding. However, and association existed between the rampant caries, the age (p < 0.001) of the child and frequency of daily consumption of sugar (p = 0.026). The odds for rampant caries was 0.63 for every year decrease in age (95% CI 0.53 to 0.76) p < 0.001. Also, the odds for rampant caries was 1.46 for every increase in frequency of daily consumption of sugar (95% CI 1.10 to 1.95) p = 0.009. However, threshold for rampant caries was established at a frequency of daily consumption of sugar greater than three times a day. At this threshold, the risk for rampant caries increased by 0.26 when the frequency of daily consumption of sugar is greater than 3 times a day (95% CI 0.023 to 0.50) p = 0.032. Children who report consuming sugary diet more than three times a day should be managed as a high risk patient for rampant caries.


Histocytologic diagnosis of neuroendocrine tumors in the liver: a retrospective study of 23 cases.
A series of 23 needle biopsies of neuroendocrine tumors occurring in the liver is described. Aspirate smears, core biopsies, and touch preparations were examined. Eighteen of the 23 patients had been previously diagnosed: 9 patients had been correctly identified as having a neuroendocrine tumor, and 9 patients had been originally misdiagnosed. Five of the patients in this series had no previously identified neoplasia. Immunohistochemical staining confirmed the neuroendocrine nature of the tumors in each of the cases. On the basis of cytomorphology, these cases were subtyped as either round cell type, spindle cell type, or polygonal cell type. The polygonal cell type of neuroendocrine tumor, as well as rare examples of the round cell type, demonstrated features similar to well-differentiated hepatocellular carcinoma and adenocarcinomas, and may present a diagnostic dilemma. Characteristic cytologic attributes of the polygonal cell type of neuroendocrine tumor which aid in its distinction from well-differentiated hepatocellular carcinoma include eccentrically located "plasmacytoid" nuclei and cellular discohesion. Findings on core needle biopsy which further identify the neuroendocrine tumors are thick fibrous stroma or small "nests" of tumor cells. The additional use of immunohistochemical staining provides reliable evidence of the cell of origin in confusing cases. Attention to these considerations will aid in the cytologic diagnosis of neuroendocrine tumors.


Intra-articular injection of epigallocatechin (EGCG) crosslinks and alters biomechanical properties of articular cartilage, a study via nanoindentation.
Osteoarthritis and rheumatoid arthritis are debilitating conditions, affecting millions of people. Both osteoarthritis and rheumatoid arthritis degrade the articular cartilage (AC) at the ends of long bones, resulting in weakened tissue prone to further damage. This degradation impairs the cartilage's mechanical properties leading to areas of thinned cartilage and exposed bone which compromises the integrity of the joint. No preventative measures exist for joint destruction. Discovering a way to slow the degradation of AC or prevent it would slow the painful progression of the disease, allowing millions to live pain-free. Recently, that the articular injection of the polyphenol epigallocatechin-gallate (EGCG) slows AC damage in an arthritis rat model. It was suggested that EGCG crosslinks AC and makes it resistant to degradation. However, direct evidence that intraarticular injection of EGCG crosslinks cartilage collagen and changes its compressive properties are not known. The aim of this study was to investigate the effects of intraarticular injection of EGCG induced biomechanical properties of AC. We hypothesize that in vivo exposure EGCG will bind and crosslink to AC collagen and alter its biomechanical properties. We developed a technique of nano-indentation to investigate articular cartilage properties by measuring cartilage compressive properties and quantifying differences due to EGCG exposure. In this study, the rat knee joint was subjected to a series of intraarticular injections of EGCG and contralateral knee joint was injected with saline. After the injections animals were sacrificed, and the knees were removed and tested in an anatomically relevant model of nanoindentation. All mechanical data was normalized to the measurements in the contralateral knee to better compare data between the animals. The data demonstrated significant increases for reduced elastic modulus (57.5%), hardness (83.2%), and stiffness (17.6%) in cartilage treated with injections of EGCG normalized to those treated with just saline solution when compared to baseline subjects without injections, with a significance level of alpha = 0.05. This data provides evidence that EGCG treated cartilage yields a strengthened cartilage matrix as compared to AC from the saline injected knees. These findings are significant because the increase in cartilage biomechanics will translate into resistance to degradation in arthritis. Furthermore, the data suggest for the first time that it is possible to strengthen the articular cartilage by intraarticular injections of polyphenols. Although this data is preliminary, it suggests that clinical applications of EGCG treated cartilage could yield strengthened tissue with the potential to resist or compensate for matrix degradation caused by arthritis.


'Big Bang' tomography as a new route to atomic-resolution electron tomography.
Until now it has not been possible to image at atomic resolution using classical electron tomographic methods, except when the target is a perfectly crystalline nano-object imaged along a few zone axes. The main reasons are that mechanical tilting in an electron microscope with sub-ångström precision over a very large angular range is difficult, that many real-life objects such as dielectric layers in microelectronic devices impose geometrical constraints and that many radiation-sensitive objects such as proteins limit the total electron dose. Hence, there is a need for a new tomographic scheme that is able to deduce three-dimensional information from only one or a few projections. Here we present an electron tomographic method that can be used to determine, from only one viewing direction and with sub-ångström precision, both the position of individual atoms in the plane of observation and their vertical position. The concept is based on the fact that an experimentally reconstructed exit wave consists of the superposition of the spherical waves that have been scattered by the individual atoms of the object. Furthermore, the phase of a Fourier component of a spherical wave increases with the distance of propagation at a known 'phase speed'. If we assume that an atom is a point-like object, the relationship between the phase and the phase speed of each Fourier component is linear, and the distance between the atom and the plane of observation can therefore be determined by linear fitting. This picture has similarities with Big Bang cosmology, in which the Universe expands from a point-like origin such that the distance of any galaxy from the origin is linearly proportional to the speed at which it moves away from the origin (Hubble expansion). The proof of concept of the method has been demonstrated experimentally for graphene with a two-layer structure and it will work optimally for similar layered materials, such as boron nitride and molybdenum disulphide.


Six-minute walking test to assess exercise tolerance and cardiorespiratory responses during training program in children with congenital heart disease.
This study assessed the exercise tolerance and the cardiorespiratory responses to a training program by the six-minute walk test (6'WT) in children with congenital heart disease (CHD). Seventeen cardiac and 14 healthy children performed maximal cardiopulmonary exercise test (CPET) and 6'WT. Reliability of 6'WT was assessed in all subjects (test-retest) by Bland-Altman plots. Cardiac subjects were randomly divided in training (T-CHD) and control groups (C-CHD). T-CHD underwent an individualized training exercise at the ventilatory threshold (VT) intensity during 12 weeks. We found that the 6'WT is a reliable and reproducible test. CHD children walked a lower distance than healthy children before training (472.5 +/- 18.1 vs. 548.8 +/- 7.7 m, respectively, p < 0.001). Likewise, power output, oxygen uptake (V.O (2)), and heart rate (HR) at the maximum and the VT levels, were significantly lower in patients (p < 0.001). After training, a significant improvement of walking distance (WD) was shown in T-CHD (529.6 +/- 15.3 vs. 467.7 +/- 17.1 m, p < 0.001). The power output, VO2, HR, and V.E increased slightly (6 to 10 %, p > 0.05) at peak exercise and significantly at ventilatory threshold level (p < 0.05) in T-CHD. Significant relationships between WD and VO2max as well as VO2 at VT were founded (p < 0.05). We concluded that the 6'WT is a useful and reliable tool in the assessment and follow-up of functional capacity during rehabilitation program in children with CHD.


Starving neurons show sex difference in autophagy.
Sex-dependent differences in adaptation to famine have long been appreciated, thought to hinge on female versus male preferences for fat versus protein sources, respectively. However, whether these differences can be reduced to neurons, independent of typical nutrient depots, such as adipose tissue, skeletal muscle, and liver, was heretofore unknown. A vital adaptation to starvation is autophagy, a mechanism for recycling amino acids from organelles and proteins. Here we show that segregated neurons from males in culture are more vulnerable to starvation than neurons from females. Nutrient deprivation decreased mitochondrial respiration, increased autophagosome formation, and produced cell death more profoundly in neurons from males versus females. Starvation-induced neuronal death was attenuated by 3-methyladenine, an inhibitor of autophagy; Atg7 knockdown using small interfering RNA; or L-carnitine, essential for transport of fatty acids into mitochondria, all more effective in neurons from males versus females. Relative tolerance to nutrient deprivation in neurons from females was associated with a marked increase in triglyceride and free fatty acid content and a cytosolic phospholipase A2-dependent increase in formation of lipid droplets. Similar sex differences in sensitivity to nutrient deprivation were seen in fibroblasts. However, although inhibition of autophagy using Atg7 small interfering RNA inhibited cell death during starvation in neurons, it increased cell death in fibroblasts, implying that the role of autophagy during starvation is both sex- and tissue-dependent. Thus, during starvation, neurons from males more readily undergo autophagy and die, whereas neurons from females mobilize fatty acids, accumulate triglycerides, form lipid droplets, and survive longer.


The cost of childhood immunisation in general practice.
To estimate the cost, to general practices in the Wellington Immunisation Network, of the audit process of recalling and immunising children according to the New Zealand Immunisation Schedule. Practices recorded all clinical and clerical time spent on immunisation as well as the materials used throughout one audit cycle. Staff time and materials were costed directly. Practice overheads were apportioned to immunisation according to the actual time spent on each of the tasks relating to immunisation relative to the total staff hours at the practice. The average cost of immunising a child who attended a non-capitated practice after a single reminder or recall was $15.15. The cost to the practice after taking the practice nurse subsidy and GMS into account was $8.51. The cost of immunising children who were not immunised at the first recall increased in proportion to the number of recall reminders. The annual average cost of immunisation to practices in the study exceeded the revenue obtained from the Immunisation Benefit. Overall, given the frequency of recall reminders, there was a net cost to practices for childhood immunisation after deducting the current immunisation benefit rate of $9.78 excluding GST. Thus, the practices in this study made a "loss" in carrying out childhood immunisations.


The contribution of adolescent growth to shorter adult statures among girls of Chinese ancestry.
Researchers have hypothesized that shorter mean statures at onset of the adolescent spurt and more rapid skeletal maturation during adolescence among East Asian populations contribute significantly to their shorter mean adult statures compared to European populations. The goal of this study is to use longitudinal height growth data modeled with the Preece-Baines Model 1 (PB1) function to examine the relative importance of adolescent growth to population differences in adult stature. Biological variables for 303 Chinese females from the Da-an District in Taipei, Taiwan, are compared with those for 23 British and 69 Euro-American females. Results indicate that 96% of the difference in adult statures (4.37 cm) between the British and urban Chinese may be attributed to the greater amount of time available to the British for preadolescent growth. Growth during adolescence between these two groups is not significantly different. When comparisons are made with the Euro-American sample from Berkeley, California, only about 26% of the difference in adult stature (6.96 cm) may be attributed to the later age at take-off. Much (53.2%) of the difference has already accrued by take-off among the Chinese. Small, but statistically significant differences in adolescent growth also exist. When considered within the context of other longitudinal studies of European and American females, this study gives only modest support to the hypothesis that more rapid skeletal maturation leads to reduced amounts of adolescent growth among East Asian populations. Interestingly, though, population differences do appear to exist in the relationship between the relative timing of an individual's growth spurt and spurt intensity. Am. J. Hum. Biol. 10:735-746, 1998. © 1998 Wiley-Liss, Inc.


False-positive finding in FDG-PET in a patient with seminoma and sarcoidosis.
A 33-year-old white man with testicular cancer (seminoma) stage IIIa showed a response in mediastinal lymph node metastases after chemotherapy. However, the tumor markers increase slightly, and therefore an (18)F-fluorodeoxyglucose (FDG) positron emission tomography/computed tomography was performed. Multiple FDG avid lesions were found in lymph node, liver and bone. The standardized uptake value (SUV) was discrete increase with a value of 2·8 to 5·7, which is more typical for inflammatory disease than a malignancy. The histological findings in a mediastinal lymph node reported a sarcoidosis, and therefore all lesions were diagnosed as a sarcoidosis. The one-year follow-up showed no further increase in tumor markers and no evidence for metastases.


Investigation of the Mechanical Properties of Iron Tailings Concrete Subjected to Dry-Wet Cycle and Negative Temperature.
This research investigates the effects of iron tailings content on the mechanical properties and durability of concrete under dry-wet cycling and negative temperature conditions (-10 °C), where iron tailings replace river sand at rates of 0%, 10%, 20%, and 30%. A variety of tests were conducted on the iron tailings concrete, including compressive strength, flexural strength, splitting tensile strength, mass loss, and relative dynamic modulus, and its pore characteristics were analyzed using low-field nuclear magnetic resonance (NMR) experiments. The results reveal that when 20% of the river sand was replaced with iron tailings, the concrete achieved optimal splitting strength, compressive strength, and flexural strength at 28 days, improving by 0.46 MPa, 3.14 MPa, and 0.41 MPa, respectively, compared to conventional concrete. Furthermore, the concrete containing this proportion of iron tailings demonstrated superior mechanical properties and durability in both negative temperature conditions and dry-wet cycling experiments. Due to the excellent physical and chemical properties of iron tailings, they enhance the performance of concrete when incorporated in appropriate quantities. The fine granularity of iron tailings helps to compensate for the granularity defects in concrete aggregates by filling internal voids, optimizing the pore structure, and improving the concrete's density and integrity. This enhances the concrete's mechanical properties and its resistance to external solutions and harmful ion penetration. Additionally, the active substances in iron tailings promote the hydration reaction of cement, leading to the formation of an increased amount of C-S-H gel and other hydration products in the cement system.


Patients' perceptions of telehealth services for outpatient treatment of substance use disorders during the COVID-19 pandemic.
The rapid scale-up of telehealth services for substance use disorders (SUDs) during the COVID-19 pandemic presented a unique opportunity to investigate patient experiences with telehealth. This study examined patient perceptions of telehealth in an outpatient SUD treatment program offering individual therapy, group therapy, and medication management. Two hundred and seventy adults receiving SUD outpatient treatment were eligible to complete a 23-item online survey distributed by clinicians; 58 patients completed/partially completed the survey. Data were summarized with descriptive statistics. Participants were predominately male, White, and well-educated. The majority (86.2%) were "very satisfied" or "satisfied" with the quality of telehealth care. "Very satisfied" ratings were highest for individual therapy (90%), followed by medication management (75%) and group therapy (58%). Top reasons for liking telehealth included the ability to do it from home (90%) and not needing to spend time commuting (83%). Top reasons for disliking telehealth were not connecting as well with other members in group therapy (28%) and the ability for telehealth to be interrupted at home or work (26%). Telehealth visits were a satisfactory treatment modality for most respondents receiving outpatient SUD care, especially those engaging in individual therapy. Challenges remain for telehealth group therapy. This is the first study examining patients' perceptions of telehealth for outpatient SUD treatment during the COVID-19 pandemic by treatment service type. Importantly, while many participants found telehealth more accessible than in-person treatment, there was variability with respect to the preferred mode of treatment delivery.


Hemodialysis and L-arginine, but not D-arginine, correct renal failure-associated endothelial dysfunction.
In end-stage renal failure (ESRF) symptomatic hemodialysis-related hypotension may prevent effective provision of renal replacement therapy. Endogenous inhibitors of nitric oxide synthase accumulate in ESRF and are cleared by dialysis. We, therefore, hypothesised that removal of these inhibitors by hemodialysis would increase endothelial nitric oxide generation and promote venodilation. In vivo responses of norepinephrine preconstricted dorsal hand veins to locally active doses of acetylcholine (an activator of nitric oxide synthase) and glyceryl trinitrate (GTN; a nitric oxide donor) were examined in patients undergoing maintenance hemodialysis for ESRF and in healthy age- and sex-matched controls. Patient studies were undertaken before and after dialysis. Studies before dialysis were repeated with co-infusion of either L-arginine or its inactive enantiomer D-arginine. Venodilation in response to acetylcholine was impaired before, and corrected by, dialysis whereas venodilation to GTN was similar before and after dialysis. Venodilation in response to acetylcholine before dialysis was restored by co-infusion of L- but not D-arginine. Therefore, patients with ESRF undergoing hemodialysis have impaired acetylcholine-mediated venodilation consistent with the accumulation in ESRF of functionally important inhibitors of nitric oxide synthase that are cleared by dialysis.


Analysis of globin gene structure in patients with beta thalassemia by restriction endonuclease mapping.
Twenty-six DNA samples from individuals either heterozygous or homozygous for beta thalassemia were analyzed by restriction endonuclease digestion, agarose gel electrophoresis, and Southern blot analysis to define DNA fragments containing portions or all of the beta globin gene. A total of twenty-seven genes affected by a beta thalassemia mutation and twenty-seven genes affected by a beta thalassemia mutation and twenty-two normal beta globin genes were examined in Italian, Greek, or Asian individuals. With all four restriction endonucleases used, the fragments generated from DNA of thalassemic individuals were identical to those found in DNA from normal. Thus, gross rearrangement or deletion within the genomic region containing the beta globin gene is not characteristic of mutations which cause a thalassemia. A third patient homozygous for pancellular hereditary persistence of fetal hemoglobin was shown to have complete deletion of the delta and beta globin genes.


Low Adherence to Society Guidelines for the Management of Helicobacter Pylori Among Pediatric Gastroenterologists.
The revised ESPGHAN/NASPGHAN 2016 guideline on the diagnosis and management of Helicobacter pylori (H pylori) infection discourages a "test and treat" strategy. Instead, upper endoscopy (EGD) is recommended when a valid clinical indication is present. Likewise, new treatment recommendations for first-line therapies strongly encourage obtaining antimicrobial susceptibility before treatment. We conducted this study to assess the effect of revised guidelines on clinical practice at our center. Retrospective chart review of patients with H pylori infection diagnosis either by serology, stool antigen test, urea breath test, or EGD at Boston Children's Hospital between January 2013 and July 2019. We compared demographic and clinical data between initial guideline and 2016 revision (January 2013 to November 2016) and period after revised guideline (December 2016 to July 2019). Two hundred and fifty-six patients were included. EGD was the initial diagnostic test in 49% (50/103, prerevised guideline) and in 52% (79/153, postrevised guideline). Biopsy culture was sent in 3% of patients for both periods (3/103 and 4/153, respectively). PPI-clarithromycin-amoxicillin triple therapy was the most common regimen in both periods. Clarithromycin use was lower in postrevised guideline period (P = 0.003) whereas the opposite was noted for metronidazole and tetracycline (P = 0.009 and P = 0.02, respectively). There was no significant difference in eradication rate between periods (86% vs 81%). Low adherence to the updated H pylori guideline recommendations was observed among pediatric gastroenterologists at our center with low use of gastric biopsy culture. The guideline revision was associated with avoidance of clarithromycin use as a second-line therapy, but no change in eradication rates. Future interventions should address the importance of obtaining gastric biopsy culture for antibiotic susceptibilities to guide therapy.


Diabetes mellitus and the exocrine pancreas.
Diabetes and carbohydrate intolerance can occur in pancreatitis. Although one-half of patients with acute pancreatitis will have some evidence of glucose intolerance during their acute illness, few will require insulin administration on either a short- or long-term basis. The diabetes seen in acute pancreatitis is likely due to a combination of factors, including alerted insulin secretion, increased glucagon release, and decreased glucose utilization by the liver and peripheral tissue. Chronic pancreatitis is often associated with diabetes mellitus, with the incidence as high as 70 percent when pancreatic calcification is present. These patients tend to be very sensitive to the effects of insulin and hypoglycemia. This is probably secondary to concurrent hepatic disease, malnutrition, and a relative decrease in glucagon reserves. The diabetes seen in chronic pancreatitis is associated with decreased insulin production. Finally, although the endocrine pancreas may influence the exocrine gland through a portal system, primary diabetes mellitus probably does not result in clinically significant alterations in pancreatic exocrine function.


Hospital volumes for common pediatric specialty operations.
To describe hospital volumes for common pediatric specialty operations, to evaluate hospital and patient characteristics associated with operations performed at a low-volume hospital, and to evaluate outcomes with hospital volume. Retrospective cohort using the Kids' Inpatient Database 2003. Discharges from 3438 hospitals in 36 states from 2003. Children aged 0 to 18 years undergoing ventriculoseptal defect surgery (n = 2301), tracheotomy (n = 2674), ventriculoperitoneal shunt placement (n = 3378), and posterior spinal fusion (n = 4002). Hospital volume. In-hospital mortality and postoperative complications. For tracheotomy and posterior spinal fusion, at least one fourth of the hospitals performed only 1 operation for children aged 0 to 18 years in 2003. For these same operations, at least half of hospitals treated 4 or fewer cases per year. For all operations, discharges from low-volume hospitals were less likely to be from children's or teaching hospitals compared with discharges from higher-volume hospitals. For tracheotomy, children were less likely to experience postoperative complications in high-volume hospitals compared with low-volume hospitals (odds ratio, 0.48; 95% confidence interval, 0.21-1.09). Many children undergoing common pediatric specialty operations had these procedures performed in low-volume hospitals. Low-volume hospitals were less likely to be children's or teaching hospitals. Children undergoing tracheotomy experienced higher rates of complications in low-volume hospitals. Further research is needed to identify the reasons why so many children have these operations performed in low-volume hospitals.


Liver inter-organelle membrane contact sites revealed by serial section electron tomography.
Inter-organelle membrane contact sites (MCSs) are defined as areas of close proximity between the membranes of two organelles (10-80nm). They have been implicated in many physiological processes such as Ca++, lipids or small molecules transfer, organelles biogenesis or dynamic and have an important role in many cellular processes such as apoptosis, autophagy, and signaling. Since the distance and the extent of these contacts are in the nanometer range, high resolution techniques are ideal for imaging these structures. It is for this reason that transmission electron microscopy (TEM) has been considered the gold standard for MCSs visualization and the first technique that described them. However, often TEM analysis is limited to 2D lacking information on the 3D association between the organelles involved in MCSs. To fully describe the complex architecture of MSCs and to unveil their role in cellular physiology a 3D analysis is required. This chapter provides a method for the analysis of MCSs using serial section electron tomography (ssET), a technique able to reconstruct in 3D at nanometer resolution cellular and subcellular structures. By applying this procedure, it was possible to elucidate the role of the contacts between Endoplasmic Reticulum (ER) and other organelles in liver lipid metabolism.


Production of env-deficient rous sarcoma virus (RSV) early after infection.
Production of defective virus particles during the early stage of Rous sarcoma virus (RSV) infection of chicken embryo fibroblasts (CEF) was examined. RSV harvested 2 days post infection (2pi) had 10 to 30 times lower specific infectivity (focus forming units/unit reverse transcriptase activity) than 5pi harvest. Virus particles produced on day 2 contained less env proteins than particles harvested on day 5. The amount of other viral proteins was equal in particles harvested on day 2 and day 5. Analysis of infected cells revealed that these cells synthesized less env proteins on day 2 than on day 5. RSV RNA in infected cells was spliced normally on day 2. Infection at a low multiplicity of infection (moi) prolonged the production of defective particles. When infection was initiated by a low moi (0.01), particles harvested on day 5 had the same characteristics as 2pi particles after infection with a high moi (1.0). We conclude that the low infectivity of early harvest is due to the reduced amount of env proteins in virus particles, which is a consequence of the reduced env protein synthesis.


Cost-effectiveness analysis of solifenacin versus oxybutynin immediate-release in the treatment of patients with overactive bladder in the United Kingdom.
To carry out a cost-utility analysis comparing initial treatment with solifenacin 5 mg/day vs oxybutynin immediate-release (IR) 15 mg/day for the treatment of patients with overactive bladder (OAB) from the perspective of the U.K. National Health Service (NHS). A Markov model with six health states was developed to follow a cohort of OAB patients treated with either solifenacin or oxybutynin during a 1-year period. Costs and utilities were accumulated as patients transited through the health states in the model and a drop-out state. Some of the solifenacin patients were titrated from 5 mg to 10 mg/day at 8 weeks. A proportion of drop-out patients were assumed to continue treatment with tolterodine ER. Utility values were obtained from a Swedish study and pad use was based on a multinational clinical trial. Adherence rates for individual treatments were derived from a U.K. database study. For pad use and utility values, the drop-out state was split between those patients who were no longer receiving treatment and those on second-line therapy. Patients on second-line therapy who drop-out were referred for a specialist visit. Results were expressed in terms of incremental cost-utility ratios. Total annual costs for solifenacin and oxybutynin were £504.30 and £364.19, respectively. First-line drug use represents 49% and 4% of costs and pad use represent 23% and 40% of costs for solifenacin and oxybutynin, respectively. Differences between cumulative utilities were small but were greater for solifenacin (0.7020 vs. 0.6907). The baseline incremental cost-effectiveness ratio was £12,309/QALY. Under the baseline assumptions, solifenacin would appear to be cost-effective with an incremental cost-utility of less than £20,000/QALY. However, small differences in utility between the alternatives and the large number of drop-outs means that the results are sensitive to small adjustments in the values of utilities assigned to the drop-out state.


Cytotoxic Escherichia coli strains encoding colibactin isolated from immunocompromised mice with urosepsis and meningitis.
Immune-compromised mouse models allow for testing the preclinical efficacy of human cell transplantations and gene therapy strategies before moving forward to clinical trials. However, CRISPR/Cas9 gene editing of the Wsh/Wsh mouse strain to create an immune-compromised model lacking function of Rag2 and Il2rγ led to unexpected morbidity and mortality. This warranted an investigation to ascertain the cause and predisposing factors associated with the outbreak. Postmortem examination was performed on 15 moribund mice. The main lesions observed in these mice consisted of ascending urogenital tract infections, suppurative otitis media, pneumonia, myocarditis, and meningoencephalomyelitis. As Escherichia coli strains harboring polyketide synthase (pks) genomic island were recently isolated from laboratory mice, the tissue sections from the urogenital tract, heart, and middle ear were subjected to E. coli specific PNA-FISH assay that revealed discrete colonies of E. coli associated with the lesions. Microbiological examination and 16S rRNA sequencing confirmed E. coli-induced infection and septicemia in the affected mice. Further characterization by clb gene analysis and colibactin toxicity assays of the pks+ E. coli revealed colibactin-associated cytotoxicity. Rederivation of the transgenic mice using embryo transfer produced mice with an intestinal flora devoid of pks+ E. coli. Importantly, these barrier-maintained rederived mice have produced multiple litters without adverse health effects. This report is the first to describe acute morbidity and mortality associated with pks+ E. coli urosepsis and meningitis in immunocompromised mice, and highlights the importance of monitoring and exclusion of colibactin-producing pks+ E. coli.


Ceramide-1-phosphate blocks apoptosis through inhibition of acid sphingomyelinase in macrophages.
It was reported previously that ceramide-1-phosphate (Cer-1-P) is mitogenic for fibroblasts (Gómez-Muñoz, A., P. A. Duffy, A. Martin, L. O'Brien, H-S. Byun, R. Bittman, and D. N. Brindley. 1995. Mol. Pharmacol. 47: 883-889; Gómez-Muñoz, A., L. M. Frago, L. Alvarez, and I. Varela-Nieto. 1997. Biochem. J. 325: 435-440). We now show that Cer-1-P prevents cell death in bone-marrow-derived macrophages (BMDMs) after withdrawal of macrophage colony-stimulating factor (M-CSF). Removal of M-CSF is known to induce apoptosis in these cells. Cer-1-P blocked activation of the caspase-9/caspase-3 pathway and prevented DNA fragmentation, indicating that the enhancement of cell survival was due to inhibition of apoptosis. M-CSF deprivation resulted in activation of acid sphingomyelinase (A-SMase), increased ceramide levels, and a decrease in intracellular Cer-1-P. Exogenously added Cer-1-P inhibited A-SMase in intact BMDMs at concentrations that also prevented apoptosis. Cer-1-P also inhibited A-SMase in cell homogenates, suggesting a possible direct physical interaction of Cer-1-P with the enzyme. In conclusion, these data demonstrate that Cer-1-P blocks apoptosis in BMDMs through inhibition of A-SMase, thereby reducing ceramide generation. This adds a new dimension to the understanding of the metabolic interrelationship of ceramides and Cer-1-P, and shows how altering the balance of intracellular levels of these mediators can affect cell survival.


Pseudo-subarachnoid haemorrhage due to chronic hypoxaemia: case report and review of the literature.
The specificity of computed tomography (CT) for subarachnoid haemorrhage (SAH) is very high. However, physicians should be aware of rare false positive findings, also referred to as "pseudo-SAH". We present an unusual case in which such a finding was caused by chronic hypoxaemia. A 37-year-old male patient presented with headaches. His CT-scan showed multiple confluent subarachnoid hyperattenuations, which mimicked SAH. However, the headache was chronic and had no features typical for SAH. The patient suffered from severe chronic hypoxaemia due to congenital heart failure. On CT-angiography diffuse intracranial vessel proliferation was found and laboratory results revealed a highly raised level of haematocrit, which had both probably developed as compensatory mechanisms. A combination of these findings explained the subarachnoid hyperdensities. Magnetic resonance imaging (MRI) showed no signs of SAH and visualized hypoxaemia in cerebral veins. A diagnosis of pseudo-SAH was made. The patient's symptoms were likely due to a secondary headache attributed to hypoxia and/or hypercapnia. Therapy was symptomatic. Severe chronic hypoxaemia should be recognised as a rare cause of pseudo-SAH. Clinical evaluation and MRI help differentiate SAH from pseudo-SAH.


Recent advances in the understanding of molecular mechanisms of cartilage degeneration, synovitis and subchondral bone changes in osteoarthritis.
Osteoarthritis (OA), the most common form of degenerative joint disease, is linked to high morbidity. It is predicted to be the single greatest cause of disability in the general population by 2030. The development of disease-modifying therapy for OA currently face great obstacle mainly because the onset and development of the disease involve complex molecular mechanisms. In this review, we will comprehensively summarize biological and pathological mechanisms of three key aspects: degeneration of articular cartilage, synovial immunopathogenesis, and changes in subchondral bone. For each tissue, we will focus on the molecular receptors, cytokines, peptidases, related cell, and signal pathways. Agents that specifically block mechanisms involved in synovial inflammation, degeneration of articular cartilage, and subchondral bone remodeling can potentially be exploited to produce targeted therapy for OA. Such new comprehensive agents will benefit affected patients and bring exciting new hope for the treatment of OA.


Portable near-infrared rapid detection of intracranial hemorrhage in Chinese population.
Secondary brain injury is the main cause of mortality from traumatic brain injury (TBI). One hallmark of TBI is intracranial hemorrhage, which occurs in 40-50% of severe TBI cases. Early identification of intracranial hematomas in TBI patients allows early surgical evacuation, and can reduce the case-fatality rate of TBI. Since pre-hospital care is the weakest part of Chinese emergency care, there is an urgent need for a capability to detect brain hematomas early. The purpose of this observational study was to evaluate the performance of a near infrared (NIR) based, device to screen for traumatic intracranial hematomas in Chinese population. Data was collected using the NIR device at the time of a computed tomography (CT) or magnetic resonance imaging (MRI) scan was performed to evaluate a suspected TBI. 85 patients were included in the per protocol population. Of the 85 patients, 45 were determined by CT scan to have intracranial hemorrhage. The CT and MRI scans were read by an independent neuroradiologist who was blinded to the NIR measurements. The NIR device demonstrated sensitivity of 95.6% (95% confidence intervals [CI] 83.6-99.2%) and specificity of 92.5% (CI 78.5-98%) in detecting intracranial hematomas larger than 3.5ml in volume, and that were less than 2.5cm from the surface of the brain. These results confirm in Chinese population the results of previous studies that demonstrated a NIR based device can reliably screen for intracranial hematomas that are likely to be of clinical importance.


Designing a multiroute synthesis scheme in combinatorial chemistry.
Solid-phase mix-and-split combinatorial synthesis is often used to produce large arrays of compounds to be tested during the various stages of the drug development process. This method can be represented by a synthesis graph in which nodes correspond to grow operations and arcs to beads transferred among the different reaction vessels. In this work, we address the problem of designing such a graph which maximizes the number of produced target compounds (namely, compounds out of an input library of desired molecules), given constraints on the number of beads used for library synthesis and on the number of reaction vessels available for concurrent grow steps. We present a heuristic based on a discrete search for solving this problem, test our solution on several data sets, explore its behavior, and show that it achieves good performance.


The use of phosphorus 32 in studies on Plasmodium gallinaceum. I. The development of a method for the quantitative determination of parasite growth and development in vitro.
A quantitative method for following parasite growth in vitro is described. The method is based on the determination of the degree of incorporation of phosphorus into the nucleic acids, in particular desoxypentose nucleic acids, of the parasitized cell. Phosphorus incorporation is measured by utilizing the radioactive isotope, P(32). The degree of phosphorus incorporation into DNA was found to be closely correlated with estimates of parasite growth based on stained film examinations of intact cell preparations. The isotope was likewise found to be incorporated into the DNA of preparations in which the host cell had been disrupted by lysis and the extent of this incorporation also paralleled the impressions gained from smear examinations. It is suggested that this method provides a sensitive and objective method of studying parasite growth.