Toward Development of Novel Remote Ultrasound Robotic System Using Soft Robotics Technology.
This paper reports on the development of a novel soft robotic system for remote ultrasound applications. Direct contact of the ultrasound probe with the patient's body represents a safety risk and therefore control of the probe's positioning and applied force is a crucial task. The proposed robot uses a passive control system that provides safe interaction between the robot and the patient by leveraging soft robotics technology. The soft robot's structure can be considered as a nonlinear spring which can be designed to exert a safe force within the robot's workspace to guarantee the safety of human-robot interaction. The literature suggests that effective ultrasound imaging of both the heart and abdomen requires six degrees-of-freedom. These degrees-of-freedom consist of three translational motions, which are achieved using a novel hybrid soft cable-driven parallel robot, and three wrist motions, which is based on a universal joint design. The experimental results show that the robot can achieve all these six degrees-of-freedom, and its blocking force can be engineered to generate a uniform force within the workspace.


Comparative Genomics Reveals Evolution of a Beak Morphology Locus in a High-Altitude Songbird.
The Ground Tit (Pseudopodoces humilis) has lived on the Qinghai-Tibet Plateau for ∼5.7 My and has the highest altitudinal distribution among all parids. This species has evolved an elongated beak in response to long-term selection imposed by ground-foraging and cavity-nesting habits, yet the genetic basis for beak elongation remains unknown. Here, we perform genome-wide analyses across 14 parid species and identify 25 highly divergent genomic regions that are significantly associated with beak length, finding seven candidate genes involved in bone morphogenesis and remolding. Neutrality tests indicate that a model allowing for a selective sweep in the highly conserved COL27A1 gene best explains variation in beak length. We also identify two nonsynonymous fixed mutations in the collagen domain that are predicted to be functionally deleterious yet may have facilitated beak elongation. Our study provides evidence of adaptive alleles in COL27A1 with major effects on beak elongation of Ps. humilis.


Identification of male heterogametic sex-determining regions on the Atlantic herring Clupea harengus genome.
The sex determination system of Atlantic herring Clupea harengus L., a commercially important fish, was investigated. Low coverage whole-genome sequencing of 48 females and 55 males and a genome-wide association study revealed two regions on chromosomes 8 and 21 associated with sex. The genotyping data of the single nucleotide polymorphisms associated with sex showed that 99.4% of the available female genotypes were homozygous, whereas 68.6% of the available male genotypes were heterozygous. This is close to the theoretical expectation of homo/heterozygous distribution at low sequencing coverage when the males are factually heterozygous. This suggested a male heterogametic sex determination system in C. harengus, consistent with other species within the Clupeiformes group. There were 76 protein coding genes on the sex regions but none of these genes were previously reported master sex regulation genes, or obviously related to sex determination. However, many of these genes are expressed in testis or ovary in other species, but the exact genes controlling sex determination in C. harengus could not be identified.


Expression and amplification of engineered mouse dihydrofolate reductase minigenes.
We constructed mouse dihydrofolate reductase (DHFR) minigenes (dhfr) that had 1.5 kilobases of 5' flanking sequences and contained either none or only one of the intervening sequences that are normally present in the coding region. They were greater than or equal to 3.2 kilobase long, about one-tenth the size of the corresponding chromosomal gene. Both of these minigenes complemented the DHFR deficiency in Chinese hamster ovary dhfr-1-cells at a high frequency after DNA-mediated gene transfer. The level of DHFR enzyme in various transfected clones varied over a 10-fold range but never was as high as in wild-type Chinese hamster ovary cells. In addition, the level of DHFR in primary transfectants did not vary directly with the copy number of the minigene, which ranged from fewer than five to several hundred per genome. The minigenes could be amplified to a level of over 2,000 copies per genome upon selection in methotrexate, a specific inhibitor of DHFR. In one case, the amplified minigenes were present in a tandem array; in two other cases, a rearranged minigene plasmid and its flanking chromosomal DNA sequence were amplified. Thus, the mouse dhfr minigenes could be transcribed, expressed, and amplified in Chinese hamster ovary cells, although the efficiency of expression was generally low. The key step in the construction of these minigenes was the generation in vivo of lambda phage recombinants by overlapping regions of homology between genomic and cDNA clones. The techniques used here for dhfr should be generally applicable to any gene, however large, and could be used to generate novel genes from members of multigene families.


Detection of Male Hypogonadism in Patients with Post COVID-19 Condition.
The pathogenesis and prognosis of post COVID-19 condition have remained unclear. We set up an outpatient clinic specializing in long COVID in February 2021 and we have been investigating post COVID-19 condition. Based on the results of our earlier study showing that "general fatigue" mimicking myalgic encephalomyelitis/chronic fatigue syndrome (ME/CFS) is the most common symptom in long COVID patients, a retrospective analysis was performed for 39 male patients in whom serum free testosterone (FT) levels were measured out of 61 male patients who visited our clinic. We analyzed the medical records of the patients' backgrounds, symptoms and laboratory results. Among the 39 patients, 19 patients (48.7%) met the criteria for late-onset hypogonadism (LOH; FT < 8.5 pg/mL: LOH group) and 14 patients were under 50 years of age. A weak negative correlation was found between age and serum FT level (r = −0.301, p = 0.0624). Symptoms including general fatigue, anxiety, cough and hair loss were more frequent in the LOH group than in the non-LOH group (FT ≥ 8.5 pg/mL). Among various laboratory parameters, blood hemoglobin level was slightly, but significantly, lower in the LOH group. Serum level of FT was positively correlated with the levels of blood hemoglobin and serum total protein and albumin in the total population, whereas these interrelationships were blurred in the LOH group. Collectively, the results indicate that the incidence of LOH is relatively high in male patients, even young male patients, with post COVID-19 and that serum FT measurement is useful for revealing occult LOH status in patients with long COVID.


siRNA-mediated AMPKalpha1 subunit gene PRKAA1 silencing enhances methylmercury toxicity in HEK293 cells.
The environmental pollutant methylmercury is a potent neurotoxin. The mechanisms of toxicity and biological defense remain largely unknown. We found that inhibiting the expression of PRKAA1 (AMPKalpha1), an activated subunit of AMP-activated protein kinase (AMPK), increased susceptibility of HEK293 cells to methylmercury toxicity. Treatment of the cells with AICAR (5-aminoimidazole-4-carboxamide-1-beta-D-ribofuranoside), an AMPK activator, reduced the methylmercury toxicity. Here, we suggest for the first time that the activation (phosphorylation) of AMPK may play an important role in reducing the toxicity of methylmercury.


The cost of residential addiction treatment in public housing.
The cost of providing addiction treatment services in a variety of settings is useful information for program administrators, policy makers, and researchers. This study estimates the economic costs of providing substance abuse treatment services at Safeport, a three-phase residential treatment program focusing on addicted women living in public housing. Economic (opportunity) costs are estimated for each phase separately and for the complete program. Results indicate that the total cost of providing treatment services at Safeport in 2001 was $1,325,235. This total cost comprises $549,737 for stabilization or early abstinence (Phase I), $400,098 for relapse prevention and self-sufficiency (Phase II), and $375,400 for independent living preparation and long-term recovery (Phase III). Average daily census (number of clients/families on a typical day) was just over 11 clients/families in each phase or 34 clients/families for the entire program. The average length of stay in the three phases of the program was 12 weeks for Phase I, 20 weeks for Phase II, 18 weeks for Phase III, and 50 weeks overall. The average weekly cost per client amounted to $930 for Phase I, $677 for Phase II, $635 for Phase III, and $748 over the full program. The average cost per treatment episode amounted to $11,163 for Phase I, $13,541 for Phase II, $11,435 for Phase III, and $36,136 for the complete program. Future research should compare these cost estimates with corresponding outcome data from Safeport to perform a comprehensive economic evaluation.


Diagnosis and clinical features of trigemino-autonomic headaches.
Although severe short-lasting headaches are rare, they can be considered disabling conditions with a major impact on the quality of life of patients. These headaches can divided broadly in to those associated with autonomic symptoms, so called trigeminal autonomic cephalgias (TACs), and those with few or no autonomic symptoms. The TACs include cluster headache, paroxysmal hemicranias, hemicrania continua, and short-lasting unilateral neuralgiform headache attacks with cranial autonomic symptoms as well as short-lasting unilateral neuralgiform headache attacks with conjunctival injection and tearing syndrome. In all of these syndromes, half-sided head pain and ipsilateral cranial autonomic symptoms such as lacrimation or rhinorrhea are prominent. The paroxysmal hemicranias have, unlike cluster headaches, a very robust response to indomethacin, leading to a notion of indomethacin-sensitive headaches. The diagnosis of TACs is exclusively a clinical task. Because of the fact that cluster headache is strictly half-sided, typically involves the region around the eye and temple and often starts in the upper jaw, most patients first consult a dentist or ophthalmologist. No single instrumental examination has yet been able to define, or ensure, the correct diagnosis, or differentiate idiopathic headache syndromes. It is crucial that a trained neurologist sees these patients early so that management can be optimized and unnecessary procedures can be avoided. Although TACS are, in comparison to migraine, quite rare, they are nevertheless clinically very important for the neurologist to consider as they are easy to diagnose and the treatment is very effective in most patients.


Regional effects of an antivascular endothelial growth factor receptor monoclonal antibody on receptor phosphorylation and apoptosis in human 253J B-V bladder cancer xenografts.
Vascular endothelial growth factor (VEGF) is a key angiogenic factor in a variety of solid tumors, making it one of the most attractive therapeutic targets. VEGF promotes the proliferation, survival, and differentiation of vascular endothelial cells by stimulating autophosphorylation and activation of VEGF receptor-2 (VEGFR-2, fetal liver kinase-1, and kinase insert domain-containing receptor). We developed fluorescence-based, quantitative methods to measure total VEGFR-2, VEGFR-2 phosphorylation, apoptosis, and microvessel density and size within whole tumor cross-sections using a laser scanning cytometer. Using these methods, we characterized the effects of DC101, a blocking antibody specific for murine VEGFR-2, on orthotopic human 253J-BV bladder tumors growing in nude mice. Basal levels of receptor phosphorylation were heterogeneous, with approximately 50% of endothelial cells positive for phosphorylated VEGFR-2 at baseline. DC101 therapy resulted in a 50% decrease in overall VEGFR-2 phosphorylation and a 15-fold and 8-fold increase in endothelial cell (CD31-positive) and tumor cell apoptosis, respectively. DC101 also decreased overall tumor microvessel density, but it mostly affected smaller CD105-negative microvessels located in the periphery of the tumor. Intriguingly, anti-VEGFR-2 therapy resulted in increased mean vessel size and an increase in overall VEGFR-2 levels. Increases in total VEGFR-2 levels were localized to the tumor core and were associated with increased expression of the oxygen-sensitive transcription factor, hypoxia inducible factor-1alpha. These data suggest that VEGFR inhibitors preferentially target discrete populations of tumor endothelial cells associated with the smaller peripheral blood vessels. Thus, agents that target a single receptor (e.g., VEGFR-2) may not be sufficient to completely inhibit tumor angiogenesis.


Use of Open Claims vs Closed Claims in Health Outcomes Research.
Background: Closed claims are frequently used in outcomes research studies. Lately, the availability of open claims has increased the possibility of obtaining information faster and on a larger scale. However, because of the possibility of missing claims and duplications, these data sets have not been highly utilized in medical research. Objective: To compare frequently used healthcare utilization measures between closed claims and open claims to analyze if the possibility of missing claims in open claims data creates a downward bias in the estimates. Methods: We identified 18 different diseases using 2022 data from 2 closed claims data sets (MarketScan® and PharMetrics® Plus) and 1 open claims database (Kythera). After applying an algorithm that removes possible duplications from open claims data, we compared healthcare utilizations such as inpatient, emergency department, and outpatient use and length of stay among these 3 data sets. We applied standardized differences to compare the medians for each outcome. Results: The sample size of the open claims data sets was 10 to 65 times larger than closed claims data sets depending on disease type. For each disease, the estimates of healthcare utilization were similar between the open claims and closed claims data. The difference was statistically insignificant. Conclusions: Open claims data with a bigger sample size and more current available information provide essential advantages for healthcare outcomes research studies. Therefore, especially for new medications and rare diseases, open claims data can provide information much earlier than closed claims, which usually have a time lag of 6 to 8 months.


Rapid detection and quantification of tyrosine decarboxylase gene (tdc) and its expression in gram-positive bacteria associated with fermented foods using PCR-based methods.
In this study, PCR-based procedures were developed to detect the occurrence and quantify the expression of the tyrosine decarboxylase gene (tdc) in gram-positive bacteria associated with fermented foods. Consensus primers were used in conventional and reverse transcription PCR to analyze a collection of 87 pure cultures of lactic acid bacteria and staphylococci. All enterococci, Staphylococcus epidermidis, Lactobacillus brevis, Lactobacillus curvatus, and Lactobacillus fermentum strains and 1 of 10 Staphylococcus xylosus strains produced amplification products with the primers DEC5 and DEC3 in accordance with results of the screening plate method and with previously reported result obtained with high-performance liquid chromatography. No amplicons were obtained for tyramine-negative strains, confirming the high specificity of these new primers. A novel quantitative real-time PCR assay was successfully applied to quantify tdc and its transcript in pure cultures and in meat and meat products. This assay allowed estimation of the influence of different variables (pH, temperature, and NaCl concentration) on the tdc expression of the tyraminogenic strain Enterococcus faecalis EF37 after 72 h of growth in M17 medium. Data obtained suggest that stressful conditions could induce greater tyrosine decarboxylase activity. The culture-independent PCR procedures developed here may be used for reliable and fast detection and quantification of bacterial tyraminogenic activity without the limitations of conventional techniques.


Assessment of potential adjuvanticity of Cry proteins.
Genetically modified (GM) crops have achieved success in the marketplace and their benefits extend beyond the overall increase in harvest yields to include lowered use of insecticides and decreased carbon dioxide emissions. The most widely grown GM crops contain gene/s for targeted insect protection, herbicide tolerance, or both. Plant expression of Bacillus thuringiensis (Bt) crystal (Cry) insecticidal proteins have been the primary way to impart insect resistance in GM crops. Although deemed safe by regulatory agencies globally, previous studies have been the basis for discussions around the potential immuno-adjuvant effects of Cry proteins. These studies had limitations in study design. The studies used animal models with extremely high doses of Cry proteins, which when given using the ig route were co-administered with an adjuvant. Although the presumption exists that Cry proteins may have immunostimulatory activity and therefore an adjuvanticity risk, the evidence shows that Cry proteins are expressed at very low levels in GM crops and are unlikely to function as adjuvants. This conclusion is based on critical review of the published literature on the effects of immunomodulation by Cry proteins, the history of safe use of Cry proteins in foods, safety of the Bt donor organisms, and pre-market weight-of-evidence-based safety assessments for GM crops.


Surfactant Protein A Recognizes Outer Membrane Protein OprH on Pseudomonas aeruginosa Isolates From Individuals With Chronic Infection.
Surfactant protein A (SP-A) plays a critical role in the clearance of Pseudomonas aeruginosa from the lung. However, there is limited information about the interaction of this protein with P. aeruginosa isolates from individuals with cystic fibrosis (CF). We characterized the interplay between SP-A and a collection of isogenic sequential isolates from 7 patients with CF. We identified outer membrane protein OprH as a novel ligand for SP-A on P. aeruginosa The last-available (late) isolates from patients with CF bound significantly less SP-A than their respective first-available (early) isolates. This difference could be associated with a reduction in the expression of OprH. Binding of SP-A to OprH promoted phagocytic killing; thus, late CF isolates were at least 2-fold more resistant to SP-A-mediated killing by human macrophages than their respective early isolates. We postulate that the reduction of OprH expression is a previously unrecognized adaptation of P. aeruginosa to the lung of individuals with CF that facilitates the escape of the microorganism from SP-A-mediated phagocytic killing.


Recognition of 3D Chiral Microenvironments for Myoblast Differentiation.
Cell chirality plays a critical role in the linkage between molecular chirality and the asymmetrical biological functions of body organs. However, enantioselective interactions between cell chirality and the extracellular environment are not yet fully understood. In this study, we investigated the effects of structurally chiral extracellular microenvironments on cellular alignments and differentiations. Twisted wrinkle-shaped chiral micropatterns were prepared using biaxial and asymmetric buckling methods, wherein structural handedness was determined from the orientation of the tilt angle between the first and second microwrinkles. Myoblasts were separately cultured on two enantiomeric chiral micropatterns in a mirror-reflected shape. Cells cultured on the left-handed chiral micropatterns preferred alignments along the direction of the second microwrinkle, with a relatively deeper valley than that of the first microwrinkle. The aligned cells on the left-handed pattern showed higher differentiation rates, as assessed by fusion indices and marker protein expression levels, than those cultured on right-handed chiral micropatterns. These results suggest that myoblasts exhibit enantioselective recognition of structurally chiral microenvironments, which can promote cellular alignments and differentiation.


A unique transcriptome: 1782 positions of RNA editing alter 1406 codon identities in mitochondrial mRNAs of the lycophyte Isoetes engelmannii.
The analysis of the mitochondrial DNA of Isoetes engelmannii as a first representative of the lycophytes recently revealed very small introns and indications for extremely frequent RNA editing. To analyze functionality of intron splicing and the extent of RNA editing in I. engelmannii, we performed a comprehensive analysis of its mitochondrial transcriptome. All 30 groups I and II introns were found to be correctly removed, showing that intron size reduction does not impede splicing. We find that mRNA editing affects 1782 sites, which lead to a total of 1406 changes in codon meanings. This includes the removal of stop codons from 23 of the 25 mitochondrial protein encoding genes. Comprehensive sequence analysis of multiple cDNAs per locus allowed classification of partially edited sites as either inefficiently edited but relevant or as non-specifically edited at mostly low frequencies. Abundant RNA editing was also found to affect tRNAs in hitherto unseen frequency, taking place at 41 positions in tRNA-precursors, including the first identification of U-to-C exchanges in two tRNA species. We finally investigated the four group II introns of the nad7 gene and could identify 27 sites of editing, most of which improve base pairing for proper secondary structure formation.


Learning from product labels and label changes: how to build pharmacogenomics into drug-development programs.
The 2010 US FDA-Drug Industry Association (DIA) Pharmacogenomics (PGx) Workshop follows a series that began in 2002 bringing together multidisciplinary experts spanning regulatory authorities, medical research, healthcare and industry. This report summarizes the 'Building PGx into Labels' sessions from the workshop, which discussed the critical elements in developing PGx outcomes leading to product labels that inform efficacy and/or safety. Examples were drawn from US prescribing information, which integrated PGx knowledge into medical decisions (e.g., panitumumab, warfarin and clopidogrel). Attendees indicated the need for broader dialog and for guidelines on evidentiary considerations for PGx to be included into product labels. Also discussed was the understanding of appropriate PGx placement on labels; how to encourage adoption by medical communities of label recommendations on PGx tests; and, given the global nature of drug development, worldwide considerations including European Summary of Product Characteristics.


Effects of acute normovolaemic haemodilution and partial exchange transfusion on blood product utilization, haemostasis and haemodynamics in surgery of the thoracoabdominal aorta. A cohort study in consecutive patients.
This paper outlines the technique of acute normovolaemic haemodilution with partial exchange transfusion (ANHPET) in surgery of the thoracic and thoracoabdominal aorta. Perioperative coagulation parameters and patterns of blood product utilization observed with this technique are described and compared with results for historical controls treated without ANHPET. During thoracoabdominal aneurysm repair, acute normovolaemic haemodilution with partial exchange transfusion (ANHPET) was used to withdraw of up to 3 L of blood. This was returned to the patient at the end of the reconstruction. Albumin 5% and stored packed red cells (PRC) were used for replacement. Seven patients underwent surgery with ANHPET, and fifteen without. Univariate and multivariate analysis of variance was used to examine differences between these groups. No differences were observed between the two groups for estimated blood loss, PRC transfused, and postoperative haemoglobin concentration. The ANHPET group received fewer platelets (8 vs 22 units, p = 0.0004), cryoprecipitate (0 vs 13 units, p = 0.02), and desmopressin or epsilon-aminocaproic acid (0 of 7 vs 4 of 15 patients, p = 0.04). FFP use was not significantly different (11 vs 17 units). Postoperatively, PTT values were less prolonged (26 vs 34 sec, p = 0.05) and platelet concentration higher (218 vs 169 x 109/L, p = 0.01) in the ANHPET group. A significant reduction in the total of blood products transfused was observed in the ANHPET group (30 vs 68 units, p = 0.003). Control of hypertension was facilitated by phlebotomy so that nitroglycerine was necessary in low doses only (0.25-1.0 microgram/kg/min). ANHPET reduced blood product transfusion, improved postoperative haemostatic parameters and simplified the management of cross-clamping hypertension.


Opioid antagonist naltrexone for the treatment of pathological gambling in Parkinson disease.
Pathological gambling (PG) is a potential complication related to the treatment of Parkinson disease (PD) with dopamine agonists (DA). The cause of this disorder is unknown, but altered dopamine neurotransmission may be involved. We evaluated the efficacy and tolerability of the opioid antagonist naltrexone in the treatment of PG in PD. Our cases included 3 patients with PD who developed PG after DA treatment. Pathological gambling did not improve after reduction or discontinuation of DA. These patients responded poorly to serotonin reuptake inhibitors, whereas treatment with opioid antagonist naltrexone resulted in the remission of PG. Naltrexone treatment was well tolerated. In one patient, higher dose of naltrexone resulted in hepatic abnormalities, which resolved after dosage reduction. The opioid antagonist naltrexone could be an effective option for the treatment of PG in PD.


PERSPECTIVE: Rooster Spermatozoa Cryopreservation and Quality Assessment.
Unsuccessful rooster fertility following cryopreservation may be linked to specific changes in spermatozoa quality, which can be determined using various methods. These determinations also facilitate the design of improved freezing and thawing processes. Here, we update the current state of methodologies available for the assessment of rooster semen quality after cryopreservation. Computer-assisted sperm analyses (CASA) is one of the main systems used to analyse motion parameters of spermatozoa (total motility, progressive motility and motion parameters). Moreover, fluorescent techniques and flow cytometry can improve the assessment of various aspects of semen quality (viability, acrosome status, mitochondrial potential, lipid peroxidation, DNA damage, lipid peroxidation and cell debris removal) using specific fluorescent markers such as ethidium bromide, Yo-Pro-1, Annexin V, propidium iodide, SYBR-14, PNA, JC-1, BODIPY, acridine orange and DRAQ5. Transmission electron microscopy also yields valuable information on spermatozoa ultrastructure. The application of these techniques to rooster spermatozoa is reviewed in relation to specific freezing techniques, the effects of cryoprotective agents (CPAs) and extenders, and the determination of spermatozoa quality after cryopreservation.


Selection for avian leukosis virus integration sites determines the clonal progression of B-cell lymphomas.
Avian leukosis virus (ALV) is a simple retrovirus that causes a wide range of tumors in chickens, the most common of which are B-cell lymphomas. The viral genome integrates into the host genome and uses its strong promoter and enhancer sequences to alter the expression of nearby genes, frequently inducing tumors. In this study, we compare the preferences for ALV integration sites in cultured cells and in tumors, by analysis of over 87,000 unique integration sites. In tissue culture we observed integration was relatively random with slight preferences for genes, transcription start sites and CpG islands. We also observed a preference for integrations in or near expressed and spliced genes. The integration pattern in cultured cells changed over the course of selection for oncogenic characteristics in tumors. In comparison to tissue culture, ALV integrations are more highly selected for proximity to transcription start sites in tumors. There is also a significant selection of ALV integrations away from CpG islands in the highly clonally expanded cells in tumors. Additionally, we utilized a high throughput method to quantify the magnitude of clonality in different stages of tumorigenesis. An ALV-induced tumor carries between 700 and 3000 unique integrations, with an average of 2.3 to 4 copies of proviral DNA per infected cell. We observed increasing tumor clonality during progression of B-cell lymphomas and identified gene players (especially TERT and MYB) and biological processes involved in tumor progression.


[The study of fatty acid composition of terpug during preserve's production].
The results of research of fatty acids composition of Terpug, during the producing of preserves in souses and aspics, which are got using the method of gas liquid chromatography. It is known that during the marinating process of Terpug there is no the oxidizing destruction of lipids of muscular tissue of fish. The kind of aspic influences on fatty acids composition of preserves of Terpug. The process of ripening and storing of preserves of fish leads to the diffusion, in particular the sorption of components of aspic with fish. This process increases nutrient density, proportion W3 cem/W6 cem acids it is close to optimal meaning, testifies to high biological value of preserves.


Prevalence of the Most Common Virulence-Associated Genes among Brucella Melitensis Isolates from Human Blood Cultures in Hamadan Province, West of Iran.
Brucellosis is a widespread zoonotic disease causing considerable economic and public health problems. Despite animal vaccination, brucellosis remains endemic in some areas such as Iran, especially in the western Iranian province of Hamadan. We sought to detect some of the most common virulence-associated genes in Brucella isolated from human blood cultures to determine the prevalence of some virulence genes among Brucella isolates. Fifty-seven isolates were studied from patients with a clinical diagnosis of brucellosis who referred to the Infectious Diseases Ward of Sina Hospital in Hamadan Province, Iran, between April 2013 and July 2014. Blood samples were collected for the diagnosis of brucellosis using the BACTEC blood culture system. All of these isolates were confirmed by the bcsp31 Brucella-specific gene. We detected 11 virulence-associated genes of Brucella, namely cβg, virB, znuA, ure, bvfA, omp25, omp31, wbkA, mviN, manA, and manB, which are important for the pathogenesis of this bacterium in the intracellular environment by multiplex PCR. Totally, 149 patients with a clinical diagnosis of brucellosis were enrolled in this study. Fifty-seven (38.3%) patients had positive blood cultures. On biochemical and molecular testing, all of the isolates were Brucella melitensis. Ten of the virulence genes were detected among all of the 57 isolates, but the bvf gene was detected in 53 (93%) isolates. The high prevalence of virulence-associated genes among the Brucella isolates detected in Hamadan Province, Iran, underscores the pathogenicity of this bacterium in this region.


Automatic segmentation of white matter hyperintensities in T2-FLAIR with AQUA: A comparative validation study against conventional methods.
White matter hyperintensities (WMHs) are lesions in the white matter of the brain that are associated with cognitive decline and an increased risk of dementia. The manual segmentation of WMHs is highly time-consuming and prone to intra- and inter-variability. Therefore, automatic segmentation approaches are gaining attention as a more efficient and objective means to detect and monitor WMHs. In this study, we propose AQUA, a deep learning model designed for fully automatic segmentation of WMHs from T2-FLAIR scans, which improves upon our previous study for small lesion detection and incorporating a multicenter approach. AQUA implements a two-dimensional U-Net architecture and uses patch-based training. Additionally, the network was modified to include Bottleneck Attention Module on each convolutional block of both the encoder and decoder to enhance performance for small-sized WMH. We evaluated the performance and robustness of AQUA by comparing it with five well-known supervised and unsupervised methods for automatic segmentation of WMHs (LGA, LPA, SLS, UBO, and BIANCA). To accomplish this, we tested these six methods on the MICCAI 2017 WMH Segmentation Challenge dataset, which contains MRI images from 170 elderly participants with WMHs of presumed vascular origin, and assessed their robustness across multiple sites and scanner types. The results showed that AQUA achieved superior performance in terms of spatial (Dice = 0.72) and volumetric (logAVD = 0.10) agreement with the manual segmentation compared to the other methods. While the recall and F1-score were moderate at 0.49 and 0.59, respectively, they improved to 0.75 and 0.82 when excluding small lesions (≤ 6 voxels). Remarkably, despite being trained on a different dataset with different ethnic backgrounds, lesion loads, and scanners, AQUA's results were comparable to the top 10 ranked methods of the MICCAI challenge. The findings suggest that AQUA is effective and practical for automatic segmentation of WMHs from T2-FLAIR scans, which could help identify individuals at risk of cognitive decline and dementia and allow for early intervention and management.


Expression Profile of p53 and p21 in Large Bowel Mucosa as Biomarkers of Inflammatory-Related Carcinogenesis in Ulcerative Colitis.
Ulcerative colitis (UC) is a chronic, relapsing inflammatory bowel disease that slightly increases the risk of colorectal cancer in patients with long-standing extended disease. Overexpression of p53 and p21 in colonic epithelia is usually detected in UC patients when no dysplasia is histologically seen and it is used by pathologists as a discriminator between regenerative changes and intraepithelial neoplasia, as well as a tissue biomarker useful to predict the risk of evolution toward malignancy. We present a one-year prospective observational study including a cohort of 45 patients with UC; p53 and p21 were evaluated in epithelial cells. p53 was positive in 74 samples revealed in 5% to 90% of epithelial cells, while 63 biopsies had strong positivity for p21 in 5% to 50% of epithelial cells. Architectural distortion was significantly correlated with p53 overexpression in epithelial cells. Thus, we consider that architectural distortion is a good substitute for p53 and p21 expression. We recommend use of p53 as the most valuable tissue biomarker in surveillance of UC patients, identifying the patients with higher risk for dysplasia. Association of p21 is also recommended for a better quantification of risk and for diminishing the false-negative results.


Multimodal interactions in free game play of children with autism and a mobile toy robot.
Autism is a complex neuropsychological disorder characterized by qualitative alterations in social interaction and interpersonal communication. The aim of this study is to estimate the interaction between autistic children and a mobile toy robot during free spontaneous game play. The duration of different criteria including eye contact, touch, manipulation, and posture have been considered. The variety of interactions of children with autism and the mobile toy robot show that the children take an interest in playing with the robot. This study suggests that the mobile toy robot in an ecological situation such as free, spontaneous game play could be used as a mediator of social stimuli in order to reduce the impairment of autistic children skills related to social information understanding and interaction.


Cheese 'refinement' with whey B-vitamin removal during precipitation potentially induces temporal 'functional' dietary shortage: homocysteine as a biomarker.
Cottage cheese 'refinement' with massive B-vitamin losses (≈70-84%) through whey removal during precipitation may potentially induce an acute imbalance between protein/methionine load and temporal inadequacy/shortage of nutrients critical for their metabolism, i.e. B6 and B12. The temporal effect of cottage cheese consumption was evaluated using increased plasma homocysteine as a B-vitamin shortage marker. In a double-blind study, healthy, normal-weight (BMI = 22-27), premenopausal women aged 25-45 years were first given a methionine load (100 mg kg(-1), n = 15), then cottage cheese alone (500 g, ≈50 g protein, ≈1200 mg methionine, n = 49) at breakfast, and then with added B6 (2 mg, n = 8) and/or B6 + folate (1 mg + 200 mcg, n = 7). Plasma homocysteine was measured preprandially (t0) and then postprandially 5 h (t5) and ≥6-24 h. Cheese-induced homocysteine increased 28.7% (p ≤ 0.001), ≈60% of the free methionine response, remaining higher through ≥6-8 h. Co-supplementation with B6 reduced the Hcy increase by 45.0% (to 14.9%, p = 0.025), and that with B6 + folate reduced the Hcy increase by 72.3% (to 7.5%, p = 0.556, NS). Homocysteine increased more in participants with lower baselines (<5 μM vs. ≥5 μM, p ≤ 0.001) following cheese, ≈3-fold (54.8% vs. 18.5%) or methionine, 47.3% (266.7% vs. 181.1%). Cheese B-vitamin depletion - i.e. to B6 ≈ 2.0-4.0 μg g(-1) protein, far below women's metabolic requirement (15-20 μg g(-1)) - appeared to induce acute relative shortage compared to methionine/protein loads, exemplified by greater homocysteine increases than with other animal proteins (previous data), more so with lower baseline homocysteine. Smaller increases following re-supplementation demonstrated potential for 'functional fortification'/co-supplementation. Unnoted cheese 'refinement', like white bread, potentially induces episodic vitamin shortage effects, warranting consideration for acute/cumulative implications, alternative processing/supplementation technologies, and food combinations, especially for at-risk populations (i.e. with genetic, hormonal/gender, or aging-related predispositions), and for cardiovascular, bone, and brain health.


The perception and knowledge of cardiovascular risk factors among medical students.
To assess perceptions, knowledge, and awareness of cardiovascular disease (CVD) risk factors among medical students (freshmen and graduating students). A descriptive cross-sectional survey based on an anonymous self-administered questionnaire was conducted in 2008 on 443 medical students - 228 freshmen on their enrollment day and 214 students on the day of their final exam at the University of Zagreb School of Medicine, Croatia. The perception and knowledge of some CVD risk factors, eg, dyslipidemia, arterial hypertension, and metabolic syndrome as well as of lipid-lowering therapy important for CVD prevention was significantly better among graduating students but was still not sufficient. Only 66% of graduating students reported that they would prescribe lipid-lowering therapy to high risk patients. Disappointingly, many graduating students were smoking (30.4%) and had low-awareness of obesity as an important CVD risk factor. These results suggest an urgent need to improve medical students' knowledge of obesity and low physical activity as important CVD risk factors and of the methods for increasing low high-density lipoprotein-cholesterol and for smoking cessation. All this provides a rationale for modifying the university core curriculum to include more information concerning these issues.


Pretreatment clinical and risk correlates of substance use disorder patients with primary depression.
The current study examined the distinction between primary and secondary depression among substance use patients to test whether the primary depressed subgroup presents to treatment with a unique profile of clinical and vulnerability characteristics. The heterogeneous sample comprised 286 individuals (76% male) with alcohol and/or drug abuse or dependence (according to criteria from the Diagnostic and Statistical Manual of Mental Disorders, Third Edition, Revised) across four treatment outcome studies conducted at the alcohol research center at the Rutgers University Center of Alcohol Studies. Participants were classified as having comorbid lifetime history of primary depression (21%), secondary depression (24%), or no depression (55%). Participants in the primary depression and secondary depression groups were comparable in severity of substance use, and both of these groups had more severe substance use problems than the no-depression group. The primary depression group presented with more severe depression histories, higher levels of current depressive symptoms, and higher rates of additional Axis I comorbidity at treatment entry. In terms of vulnerability indices, the primary depression subgroup had a uniquely high family history risk for major depressive disorder; underlying personality vulnerability to depression was also evident in the primary depression group, with higher neuroticism and lower extraversion relative to secondary depression patients. The findings suggest that careful assessment of lifetime depression symptoms vis-à-vis substance use history and severity yields important information identifying the primary depression subtype of substance use patients as a group with a unique and more severely affected clinical presentation of depression and other Axis I psychopathology relative to secondary depression patients. Effectiveness of substance use interventions may be augmented with depression treatment for primary depression patients, given their more severe clinical presentation and vulnerability characteristics.


Controlling and detecting spin correlations of ultracold atoms in optical lattices.
We report on the controlled creation of a valence bond state of delocalized effective-spin singlet and triplet dimers by means of a bichromatic optical superlattice. We demonstrate a coherent coupling between the singlet and triplet states and show how the superlattice can be employed to measure the singlet-fraction employing a spin-blockade effect. Our method provides a reliable way to detect and control nearest-neighbor spin correlations in many-body systems of ultracold atoms. Being able to measure these correlations is an important ingredient in studying quantum magnetism in optical lattices. We furthermore employ a SWAP operation between atoms which are part of different triplets, thus effectively increasing their bond-length. Such a SWAP operation provides an important step towards the massively parallel creation of a multiparticle entangled state in the lattice.


Isolation of large numbers of chromosome 3-specific cosmids containing clusters of rare restriction-endonuclease sites.
We tested 519 chromosome 3-specific cosmids for the presence of rare restriction-endonuclease sites in a search for cosmids containing HTF islands. We have identified 49 cosmids (9% of those tested) that contain multiple rare restriction-endonuclease sites. The cosmids were digested with several common cutting restriction endonucleases to liberate small fragments which were tested as unique-sequence chromosome 3-specific hybridization probes and for evolutionary sequence conservation. Unique-sequence hybridization probes isolated from the cosmids were hybridized to a somatic cell hybrid deletion mapping panel to subchromosomally localize the cosmids. Fragments from many of these cosmids demonstrated conservation of sequence through evolution, and these fragments hybridize to distinct transcripts. These cosmids should therefore prove a useful resource for the identification of many chromosome 3-specific genes, in addition to having potential use as linking clones for pulsed-field gel mapping studies.


Meprin β metalloproteases associated with differential metabolite profiles in the plasma and urine of mice with type 1 diabetes and diabetic nephropathy.
Meprin metalloproteases are abundantly expressed in the brush border membranes of kidney proximal tubules and small intestines. Meprins are also expressed in podocytes and leukocytes (monocytes and macrophages). Meprins are implicated in the pathophysiology of diabetic nephropathy (DN) but underlying mechanisms are not fully understood. Single nucleotide polymophisms (SNPs) in the meprin β gene were associated with DKD in human subjects. Furthermore, meprin α and β double deficiency resulted in more severe kidney injury and higher mortality rates in mice with Streptozotocin (STZ)-induced type 1 diabetes. Identification of meprin substrates has provided insights on how meprins could modulate kidney injury. Meprin targets in the kidney include extracellular matrix (ECM) proteins, modulators of inflammation, and proteins involved in the protein kinase A (PKA) and PKC signaling pathways. The current study used a global metabolomics approach to determine how meprin β expression impacts the metabolite milieu in diabetes and DKD. Low dose STZ was used to induce type 1 diabetes in 8-week old wild-type (WT) and meprin β knockout (βKO) mice. Blood and urine samples were obtained at 4 and 8 weeks post-STZ injection. Assays for albumin, creatinine, neutrophil gelatinase-associated lipocalin (NGAL), kidney injury molecule - 1 (KIM-1), and cystatin C were used for biochemical assessment of kidney injury. Data for biomarkers of kidney injury utilized two-way ANOVA. Metabolomics data analysis utilized UPLC-QTOF MS and multivariate statistics. The number of metabolites with diabetes-associated changes in levels were significantly higher in the WT mice when compared to meprin βKO counterparts. Annotated meprin β expression-associated metabolites with strong variable importance in projection (VIP) scores play roles in lipid metabolism (LysoPC(16:1(9Z)), taurocholic acid), amino acid metabolism (indoxyl sulfate, hippuric acid), and neurotransmitter/stress hormone synthesis (cortisol, 3-methoxy-4-hydroxyphenylethylene glycolsulfate, homovanillic acid sulfate). Metabolites that associated with meprin β deficiency include; 3,5-dihydroxy-3',4'-dimethoxy-6,7-methylenedioxyflavone 3-glucuronide, pantothenic acid, and indoxyl glucuronide (all decreased in plasma). Taken together, the annotated metabolites suggest that meprin β impacts complications of diabetes such as DKD by altering distinct metabolite profiles.


The integration experience of hearing impaired elementary school students in separated and integrated school settings.
Integration experiences of hearing impaired German elementary school students in separate educational settings (n = 31) were compared with those of counterparts at the same level in integrated settings (n = 26), and evaluated in regard to psychosocial behavior, semantic-lexical abilities, and communicative skills. Analysis of questionnaire responses and intelligence subtests showed that the only difference between the samples concerned perceived well-being in school, and favored the integrated students. Other percentages of variance are explainable by school type (10%), parents' hearing status (12%), and use of bimodal communication (11%). The integrated students demonstrated a higher level of integration experience, associated with fewer psychosocial abnormalities and better communicative skills. The results indicate that educational setting is not the only factor influencing students' perceived well-being. Other contextual factors, e.g., hearing status of parents and communicative skills, have a similar positive impact on integration experiences


Recurrent presumed herpes simplex keratitis and episcleritis in keratosis follicularis (Darier's disease).
Keratosis follicularis (Darier's disease) is an autosomal dominant dermatological disorder characterised by abnormal epidermal differentiation and loss of normal cell-to-cell adhesion. Cardinal features include diffuse hyperkeratotic warty papules with scaly plaques in seborrhoeic regions with associated mucous membrane changes. Darier's disease is rare (prevalence 2.7 in 100,000), with few ocular sequelae reported: commonly dry eye with or without Sjögren's syndrome. This is the first report, to the best of our knowledge, to describe a case of recurrent herpes simplex virus (HSV) keratitis and episcleritis in a 47-year-old man suffering from Darier's disease. The patient's condition predisposed him towards developing ocular complications due to several factors: impaired desmosome function leading to poor cell-to-cell adhesion in the corneal epithelium, dry eye and HSV invasion of inflamed periocular skin presumably combining to allow viral colonisation of a poorly protected cornea.


Effect of preservation of the inferior and posterior borders on recurrence of ameloblastomas of the mandible.
The purpose of this study was to evaluate the experience and results of using a rational radical conservative approach in the surgical treatment of mandibular ameloblastomas. A retrospective analysis of every patient with an ameloblastoma of the mandible treated in the Department of Oral Medicine and Oral Surgery at the Jordan University of Science and Technology between 1989 and 1999 was conducted. All were operated on by the author, who used a uniform surgical protocol in every case. All soft tissues in contact or overlying the lesion and a wide margin of cancellous bone related to the tumor were destroyed. The compact bone comprising the lower border of the mandible and the posterior border of the ascending ramus together with the nutrient periosteum attached to them were preserved, thus maintaining the continuity of the mandible and the facial contours. When teeth, soft tissues, or both were involved, they were destroyed. All patients were reviewed annually for a follow-up period of 3 to 10 years (mean, 6.5 years) by the same clinician. All ameloblastomas were located in the mandible; 21 were in the angle/molar region and the ascending ramus, and 2 cases were in the anterior region. There was no clinical or radiographic evidence of recurrence observed during the follow-up period. Ameloblastoma has a high rate of local recurrence if it is not adequately removed. The findings of this study to date suggest that the technique of rational radical conservative resection may have a place in the treatment of ameloblastoma of the mandible and is worthy of further trial.


Cross-sectional geometric analysis of a foot bone assemblage from Mangaia, Cook Islands.
This study describes a human foot bone assemblage from prehistoric Mangaia, Cook Islands in the context of diaphyseal cross-sectional strength measures. We use this sample to test the hypothesis that habitually unshod individuals who walk over rugged terrain will have stronger foot bones than a sample of habitually shod industrialized people. Specifically, we examine whether the Mangaian sample has a stronger size-adjusted metatarsal (MT) and phalangeal cross-sectional properties than the industrial sample, drawn from the Terry Collection. Contrary to expectations, residual analyses showed that most values of cross-sectional area (CA) and torsional resistance (J) of MTs 1-4 and the hallucal proximal phalanx (HPP) of the Mangaians are among those in the lower range of the Terry Collection sample. However, the bending strength ratios (Zy/Zx) of the Mangaian HPP are significantly greater than those of the Terry Collection. While characteristics such as forefoot shape variation between the sexes and among geographic populations cannot be ruled out as influential factors, cross-sectional properties of the hallucal proximal phalanges, but not the MTs, indicate terrain complexity in prehistoric populations.


Pressure-voltage trap for DNA near a solid-state nanopore.
We report the formation of a tunable single DNA molecule trap near a solid-state nanopore in an electrolyte solution under conditions where an electric force and a pressure-induced viscous flow force on the molecule are nearly balanced. Trapped molecules can enter the pore multiple times before escaping the trap by passing through the pore or by diffusing away. Statistical analysis of many individually trapped molecules yields a detailed picture of the fluctuation phenomena involved, which are successfully modeled by a one-dimensional first passage approach.


A spin-Seebeck diode with a negative differential spin-Seebeck effect in a hydrogen-terminated zigzag silicene nanoribbon heterojunction.
The spin-Seebeck effect (SSE), the central topic of spin caloritronics, provides a new direction for future low power consumption technology. To realize device applications of SSE, a spin-Seebeck diode (SSD) with a negative differential SSE is very desirable. To this end, we constructed a spin caloritronics device that was composed of a ferromagnetic double-single-hydrogen-terminated zigzag silicene nanoribbon (ZSiNR-H2-H) and an antiferromagnetic double-double-hydrogen-terminated zigzag silicene nanoribbon (ZSiNR-H2-H2). By using ab initio calculations combined with nonequilibrium Green's function technique, we found that thermally driven spin current through the heterojunction featured the SSD effect and negative differential SSE. The former originates from the asymmetrical thermal-driven conducting electrons and holes, and the latter ascribes to the thermal spin compensation effect. Their physical mechanisms are much different from the previous ones mainly relying on the spin-wave excitations in the interface between metals and magnetic insulators, supporting our study that puts forward a new route to realize the SSD with a negative differential SSE.


Prospective study of new participants in a community-based mind-body training program.
Mind-body practices such as yoga are widely popular, but little is known about how such exercises impact health-related quality of life. To measure changes in health-related quality of life associated with 3 months of mind-body training as practiced in community-based settings. Prospective cohort study. Eight centers for practice of mind-body training. One hundred ninety-four English-speaking adults who had taken no more than 10 classes at the centers prior to enrollment in the study. One hundred seventy-one (88%) returned the 3-month follow-up questionnaire. Administration of the SF-36 questionnaire at the start of training and after 3 months. At baseline, new participants in mind-body training reported lower scores than U.S. norms for 7 of 8 domains of the SF-36: mental health, role emotional, social, vitality, general health, body pain, and role physical (P <.002 for all comparisons). After 3 months of training, within-patient change scores improved in all domains (P <.0001), including a change of +15.5 (standard deviation +/-21) in the mental health domain. In hierarchical regression analysis, younger age (P=.0003), baseline level of depressive symptoms (P=.01), and reporting a history of hypertension (P=.0054) were independent predictors of greater improvement in the SF-36 mental health score. Five participants (2.9%) reported a musculoskeletal injury. New participants in a community-based mind-body training program reported poor health-related quality of life at baseline and moderate improvements after 3 months of practice. Randomized trials are needed to determine whether benefits may be generalizable to physician-referred populations.


Interstitial magnetic resonance lymphography with gadobutrol in rabbits and an initial experience in humans.
The purpose of this study was first to evaluate gadobutrol as a contrast agent for interstitial Magnetic Resonance Lymphography (MRL) in rabbits, and second, to extend the study to humans, if the initial results were satisfactory. In our experiment, gadobutrol was injected into twelve white New Zealand rabbits. In nine animals, 0.5 ml of gadobutrol was subcutaneously administered through each foot pad of the hindlegs while in the remaining three animals the agent was given in each foot of the forelegs. In four of the nine rabbits, slight local massage was applied at the site of administration. Subsequently, we proceeded to administer 5 ml (4.5 ml gadobutrol mixed with 0.5 ml hydrochloride lidocaine) into the limbs of two healthy humans. We achieved imaging of four lymph node groups (popliteal, inguinal, iliac and paraortic) in the hind-legs of the nine-rabbit group, whereas, in the forelegs of the remaining three rabbits, three lymph node groups (axillary, parasternal, mediastinal) were depicted. The flow of the contrast agent was significantly faster in the rabbits that received local massage (P<0.02). In humans, normal lymph vessels, as well as inguinal lymph nodes, were depicted in the legs. No side-effects were observed either in the rabbits or humans.


Proton MR spectroscopy correlates diffuse axonal abnormalities with post-concussive symptoms in mild traumatic brain injury.
There are no established biomarkers for mild traumatic brain injury (mTBI), in part because post-concussive symptoms (PCS) are subjective and conventional imaging is typically unremarkable. To test whether diffuse axonal abnormalities quantified with three-dimensional (3D) proton magnetic resonance spectroscopic imaging (¹H-MRSI) correlated with patients' PCS, we retrospectively studied 26 mTBI patients (mean Glasgow Coma Scale [GCS] score of 14.7), 18- to 56-year-olds and 13 controls three to 55 days post-injury. All were scanned at 3 Tesla with T1- and T2-weighted MRI and 3D ¹H-MRSI (480 voxels over 360 cm³, ∼30% of the brain). On scan day, patients completed a symptom questionnaire, and those who indicated at least one of the most common subacute mTBI symptoms (headache, dizziness, sleep disturbance, memory deficits, blurred vision) were grouped as PCS-positive. Global gray matter and white matter (GM/WM) absolute concentrations of N-acetylaspartate (NAA), choline (Cho), creatine (Cr) and myo-inositol (mI) in PCS-positive and PCS-negative patients were compared to age- and gender-matched controls using two-way analysis of variance. The results showed that the PCS-negative group (n=11) and controls (n=8) did not differ in any GM or WM metabolite level. The PCS-positive patients (n=15) had lower WM NAA than the controls (n=12; 7.0 ± 0.6 versus 7.9 ± 0.5mM; p=0.0007). Global WM NAA, therefore, showed sensitivity to the TBI sequelae associated with common PCS in patients with mostly normal neuroimaging, as well as GCS scores. This suggests a potential biomarker role in a patient population in which objective measures of injury and symptomatology are currently lacking.


[Decrease of latency of auditory evoked potentials in humans practicing transcendental meditation (author's transl].
Slow cortical auditory responses were recorded from humans practicing transcendental meditation. The potentials were computated according to Fig. 1, and the latencies of the positive peaks P1 and P2 and of the negative peaks N1 and N2 were measured as well as the areas of the on-effect FON, of the off-effect FOFF and of the DC-shift FDC during the stimulus duration. Latencies for most of the initial peaks during TM as well as during normal consciousness were significantly shorter than in a control group in a dozing state or during normal consciousness (Table 1 and Fig. 2). Certain differences were also found in regard to other parameters such as amplitudes and areas of the auditory evoked response.


Mindfulness and Imagery Enhanced Behavioral Parenting: Effectiveness Pilot of the Confident Carers Cooperative Kids Program.
Mindfulness is increasingly offered to parents of children presenting with behavioral problems, either as a stand-alone intervention, or integrated within existing behavioral parenting interventions. There is relatively modest support for mindful parenting, with small to medium effect size improvements demonstrated across child and parent outcome measures. Here we introduce a mindfulness and imagery enhanced behavioral parenting program. We argue blending mindfulness, imagery and behavioral skills could produce improved parenting engagement and perseverance, leading to stronger outcomes. Pilot data is presented from two contrasting real world clinical settings. Parents attending the 8-week Confident Carers Cooperative Kids (CCCK) group program in a university clinic setting were invited to be included in the study (n = 20). Permission was also gained to use archival data from a community organisation offering CCCK groups to parents who were at risk of child welfare involvement (n = 14). Pre- and post-intervention measures were completed across both groups on parent-reported child behavior, parent wellbeing, adaptive parenting, and mindful parenting. Parents from both groups achieved significant pre- to post-intervention improvements in child behavior problems, parent wellbeing, adaptive parenting, and mindful parenting, with large effect sizes. Larger improvements in child behavior problems were reported by parents from the community group compared with the university group. The CCCK intervention appears beneficial across child and parent outcomes, including for families most in need. A larger sample is required to replicate and extend these promising findings.


Are animals stuck in time?
People can time travel cognitively because they can remember events having occurred at particular times in the past (episodic memory) and because they can anticipate new events occurring at particular times in the future. The ability to assign points in time to events arises from human development of a sense of time and its accompanying time-keeping technology. The hypothesis is advanced that animals are cognitively stuck in time: that is, they have no sense of time and thus have no episodic memory or ability to anticipate long-range future events. Research on animals' abilities to detect time of day, track short time intervals, remember the order of a sequence of events, and anticipate future events are considered, and it is concluded that the stuck-in-time hypothesis is largely supported by the current evidence.


Barriers to accessing and receiving mental health care in Eastern Cape, South Africa.
The right to the enjoyment of the highest attainable standard of physical and mental health is enshrined in many international human rights treaties. However, studies have shown that people with mental disabilities are often marginalized and discriminated against in the fulfillment of their right to health. The aim of this study is to identify and reach a broader understanding of barriers to the right to mental health in the Eastern Cape Province in South Africa. Eleven semi-structured interviews were carried out with health professionals and administrators. The researchers used the Availability, Accessibility, Acceptability, and Quality (AAAQ) framework from the UN Committee on Economic, Social and Cultural Rights to structure and analyze the material. The framework recognizes these four interrelated and partly overlapping elements as necessary for implementation of the right to health. The study identifies eleven barriers to the enjoyment of the right to health for people with mental disabilities. Three categories of barriers relate to availability: lack of staff, lack of facilities, and lack of community services and preventive care. Four barriers relate to accessibility: lack of transport, lack of information, stigmatization, and traditional cultural beliefs of the community. Two barriers relate to acceptability: lack of cross-cultural understanding among staff and traditional cultural beliefs of staff. Finally, two barriers relate to quality: lack of properly trained staff and lack of organizational capacity. The results, in line with earlier research, indicate that the implementation of the right to health for people with mental disabilities is far from achieved in South Africa. The findings contribute to monitoring the right to mental health in South Africa through the identification of barriers to the right to health and by indicating the importance of building monitoring procedures based on the experiences and knowledge of staff involved in mental health care provision.


Neotelomeres and Telomere-Spanning Chromosomal Arm Fusions in Cancer Genomes Revealed by Long-Read Sequencing.
Alterations in the structure and location of telomeres are key events in cancer genome evolution. However, previous genomic approaches, unable to span long telomeric repeat arrays, could not characterize the nature of these alterations. Here, we applied both long-read and short-read genome sequencing to assess telomere repeat-containing structures in cancers and cancer cell lines. Using long-read genome sequences that span telomeric repeat arrays, we defined four types of telomere repeat variations in cancer cells: neotelomeres where telomere addition heals chromosome breaks, chromosomal arm fusions spanning telomere repeats, fusions of neotelomeres, and peri-centromeric fusions with adjoined telomere and centromere repeats. Analysis of lung adenocarcinoma genome sequences identified somatic neotelomere and telomere-spanning fusion alterations. These results provide a framework for systematic study of telomeric repeat arrays in cancer genomes, that could serve as a model for understanding the somatic evolution of other repetitive genomic elements.


Factors associated with mortality among hospitalized patients with COVID-19 disease treated with convalescent plasma.
The use of convalescent plasma (CP) for hospitalized patients with SARS-CoV-2 infection might be a useful option in certain settings. Soon after the outbreak of COVID-19, the National Ministry of Health of Argentina recommended the use of CP transfusion for hospitalized patients with COVID-19 disease. Between 1 June and 3 October 2020, 480 patients, excluding those on invasive mechanical ventilation (IMV), received at least one CP infusion in the province of Santa Fe. We aimed to find factors associated with mortality among this cohort of patients. The median age was 60 years (interquartile range: 49-69 years) and 320 (66.7%) were males. Most of these patients (93.75%) received a single CP infusion, 82.1% and 95.6% before day 4 and day 7 of hospitalization, respectively. Anti-SARS-CoV-2 titers were determined in the CP units administered using Elecsys Anti-SARS-CoV-2 S assay. At 28 days of follow-up, 250 patients were discharged (52.1%), 131 (27.3%) remained hospitalized without and 16 (3.3%) with oxygen requirement, 27 (5.6%) were on IMV, and 56 (11.7%) had died. In the multivariate logistic regression analysis, the factors significantly associated with 28-day mortality were (i) requirement of IMV, (ii) the administration of CP after the third day of hospitalization, (iii) age, and (iv) number of comorbidities. The qualitative and quantitative analyses of antibodies against SARS-CoV-2 in the infused CP were not associated with mortality. Our findings may imply a seemingly favorable effect of CP administration among patients with severe COVID-19 disease when infused sooner after hospitalization.IMPORTANCEThe use of convalescent plasma (CP) could be an option for patients with severe COVID-19, especially in poor-resource countries where direct antiviral drugs are not commercially available. Currently, the U.S. Food and Drug Administration limits the CP administration for outpatients and inpatients with COVID-19 who are immunocompromised and only if high levels of anti-SARS-CoV-2 antibodies are confirmed in the CP unit. Although most of the randomized clinical trials failed to show a clear-cut benefit of CP in hospitalized patients with severe COVID-19, other studies have shown that if given early in the course of the disease, it might be a useful therapeutic option. In this retrospective study, we demonstrated that early treatment (within 3 days of hospitalization) was significantly associated with reduced 28-day mortality compared with those patients treated beyond day 3. The results from our study add up to the scientific evidence on the use of CP as a relatively safe, cheap, and possibly effective therapy in certain patients suffering from severe SARS-CoV-2 infection.


Adrenal transcription regulatory genes modulated by angiotensin II and their role in steroidogenesis.
Transcription regulatory genes are crucial modulators of cell physiology and metabolism whose intracellular levels are tightly controlled to respond to extracellular stimuli. We studied transcription regulatory genes modulated by angiotensin II, one of the most important regulators of adrenal cortical cell function, and their role in adrenal steroidogenesis in H295R human adrenocortical cells. Angiotensin II-modulated transcription regulatory genes were identified with high-density oligonucleotide microarrays and the results validated by real-time RT-PCR. Cotransfection reporter assays were performed in H295R cells to analyze the role of these transcription regulatory genes in the control of the expression of 11beta-hydroxylase and aldosterone synthase, the last and unique enzymes of the glucocorticoid and mineralocorticoid biosynthetic pathways, respectively. We selected a subset of the most regulated genes for reporter plasmid studies to determine the effect on these enzymes. BHLHB2, BTG2, and SALL1 decreased expression of both enzymes, whereas CITED2, EGR2, ELL2, FOS, FOSB, HDAC5, MAFF, MITF, NFIL3, NR4A1, NR4A2, NR4A3, PER1, and VDR increased expression for both enzymes. By the ratio of aldosterone synthase to 11beta-hydroxylase expression, NFIL3, NR4A1, NR4A2, and NR4A3 show the greatest selectivity toward upregulating expression of the mineralocorticoid biosynthetic pathway preferentially. In summary, this study reports for the first time a set of transcription regulatory genes that are modulated by angiotensin II and their role in adrenal gland steroidogenesis. Abnormal regulation of the mineralocorticoid or glucocorticoid biosynthesis pathways is involved in several pathophysiological conditions; hence the modulated transcription regulatory genes described may correlate with adrenal steroidogenesis pathologies.


Proximity Dependent Biotinylation: Key Enzymes and Adaptation to Proteomics Approaches.
The study of protein subcellular distribution, their assembly into complexes and the set of proteins with which they interact with is essential to our understanding of fundamental biological processes. Complementary to traditional assays, proximity-dependent biotinylation (PDB) approaches coupled with mass spectrometry (such as BioID or APEX) have emerged as powerful techniques to study proximal protein interactions and the subcellular proteome in the context of living cells and organisms. Since their introduction in 2012, PDB approaches have been used in an increasing number of studies and the enzymes themselves have been subjected to intensive optimization. How these enzymes have been optimized and considerations for their use in proteomics experiments are important questions. Here, we review the structural diversity and mechanisms of the two main classes of PDB enzymes: the biotin protein ligases (BioID) and the peroxidases (APEX). We describe the engineering of these enzymes for PDB and review emerging applications, including the development of PDB for coincidence detection (split-PDB). Lastly, we briefly review enzyme selection and experimental design guidelines and reflect on the labeling chemistries and their implication for data interpretation.


Governing Heritable Human Genome Editing: A Textual History and a Proposal for the Future.
Heritable human genome editing (HHGE) has become a topic of intense public interest, especially since 2015. In the early 1980s, a related topic-human genetic engineering-was the subject of sustained public discussion. There was particular concern about germline genetic intervention. During the 1980s debate, an advisory committee to the Director of the National Institutes of Health (NIH)-the Recombinant DNA Advisory Committee (RAC)-agreed to provide initial public review of proposals for deliberate introduction of DNA into human beings. In 1984 and 1985, the RAC developed guidelines for research involving DNA transfer into patients. The committee also commented on the possibility of deliberately altering the human germline. We track the textual changes over time in the RAC's response to the possibility of germline genetic intervention in humans. In 2019, the NIH RAC was abolished. New techniques for genome editing, including CRISPR-based techniques, make both somatic and germline alterations much more feasible. These novel capabilities have again raised questions about oversight. We propose the creation of a new structure for the public oversight of proposals to perform HHGE. In parallel with a technical review by a regulatory agency, such proposals should also be publicly evaluated by a presidentially appointed Bioethics Advisory Commission.


Beyond disease susceptibility-Leveraging genome-wide association studies for new insights into complex disease biology.
Genetic studies in complex diseases have been highly successful, but have also been largely one-dimensional: predominantly focusing on the genetic contribution to disease susceptibility. While this is undoubtedly important-indeed it is a pre-requisite for understanding the mechanisms underlying disease development-there are many other important aspects of disease biology that have received comparatively little attention. In this review, I will discuss how existing genetic data can be leveraged to provide new insights into other aspects of disease biology, why such insights could change the way we think about complex disease, and how this could provide opportunities for better therapies and/or facilitate personalised medicine. To do this, I will use the example of Crohn's disease-a chronic form of inflammatory bowel disease that has been one of the main success stories in complex disease genetics. Indeed, thanks to genetic studies, we now have a much more detailed understanding of the processes involved in Crohn's disease development, but still know relatively little about what determines the subsequent disease course (prognosis) and why this differs so considerably between individuals. I will discuss how we came to realise that genetic variation plays an important role in determining disease prognosis and how this has changed the way we think about Crohn's disease genetics. This will illustrate how phenotypic data can be used to leverage new insights from genetic data and will provide a broadly applicable framework that could yield new insights into the biology of multiple diseases.


Multiscale recurrence quantification analysis of spatial cardiac vectorcardiogram signals.
Myocardial infarction (MI), also known as a heart attack, is a leading cause of mortality in the world. Spatial vectorcardiogram (VCG) signals are recorded on the body surface to monitor the underlying cardiac electrical activities in three orthogonal directions of the body, namely, frontal, transverse, and sagittal planes. The 3-D VCG vector loops provide a new way to study the cardiac dynamical behaviors, as opposed to the conventional time-delay reconstructed phase space from a single ECG trace. However, few, if any, previous approaches studied the relationships between cardiac disorders and recurrence patterns in VCG signals. This paper presents the recurrence quantification analysis (RQA) of VCG signals in multiple wavelet scales for the identification of cardiac disorders. The linear classification models using multiscale RQA features were shown to detect MI with an average sensitivity of 96.5% and an average specificity of 75% in the randomized classification experiments of PhysioNet Physikalisch-Technische Bundesanstalt database, which is comparable to the performance of human experts. This study is strongly indicative of potential automated MI classification algorithms for diagnostic and therapeutic purposes.


Cumulative Live Birth Rates in Low Prognosis Patients According to the POSEIDON Criteria: An Analysis of 26,697 Cycles of in vitro Fertilization/Intracytoplasmic Sperm Injection.
Objective: The POSEIDON criteria are used to stratify patients with low prognosis after assisted reproductive technology (ART) treatment. Since its introduction, there has been no large study about the prognosis of the POSEIDON population. We used the POSEIDON criteria in Chinese women who underwent repeated ART treatment and analyzed the association between POSEIDON criteria and the cumulative live-birth rate (CLBR). Methods: This was a retrospective cohort study of 62,749 women (97,388 cycles) who underwent ART treatment at the Reproductive and Genetic Hospital of CITIC-XIANGYA between January 2014 and June 2017. Among them, 19,781 (31.52%) women fulfilled the POSEIDON criteria, including 26,697 cycles. The optimal and conservative CLBRs within a complete IVF/ICSI treatment cycle were calculated, as well as the CLBRs following repeated ovarian stimulation cycles. Results: In POSEIDON groups 1, 2, 3, and 4, the optimal and conservative CLBRs of three complete consecutive in vitro fertilization (IVF)/intracytoplasmic sperm injection (ICSI) cycles were 83.87 and 66.06%, 53.67 and 37.72%, 44.24 and 27.98%, and 14.20 and 9.68%, respectively. The POSEIDON stratification [group 2: odds ratio (OR) = 2.319, 95% confidence interval (CI): 2.131-2.525, P < 0.001; group 3: OR = 1.356, 95% CI: 1.005-1.828, P = 0.046; group 4: OR = 3.525, 95% CI: 2.774-4.479, P < 0.001; all vs. group 1] and ovarian stimulation protocol [gonadotropin-releasing hormone (GnRH) antagonist protocol: OR = 1.856, 95% CI: 1.640-2.100, P < 0.001; other protocols: OR = 1.651, 95% CI: 1.155-2.361, P = 0.006; both vs. long GnRH agonist protocol] were associated with live birth in the first stimulation cycle. For the second stimulation cycle, the POSEIDON stratification (except POSEIDON group 3) and ovarian stimulation protocol were associated with live birth. A change in ovarian stimulation protocol was not associated with an improvement in the live birth rate. Conclusions: More than 30% of women who undergo IVF/ICSI treatment may be classified as low prognosis. Different reproductive outcomes were observed among the four POSEIDON groups. The most optimal outcomes after three successive cycles of IVF/ICSI treatment were observed in groups 1, 2, and 3.


Longer life spans and delayed maturation in wild-derived mice.
Nearly all the experimental mice used in aging research are derived from lineages that have been selected for many generations for adaptation to laboratory breeding conditions and are subsequently inbred. To see if inbreeding and laboratory adaptation might have altered the frequencies of genes that influence life span, we have developed three lines of mice (Idaho [Id], Pohnpei [Po], and Majuro [Ma]) from wild-trapped progenitors, and have compared them with a genetically heterogeneous mouse stock (DC) representative of the laboratory-adapted gene pool. Mean life span of the Id stock exceeded that of the DC stock by 24% (P < 0.00002), and maximal life span, estimated as mean longevity of the longest-lived 10% of the mice, was also increased by 16% (P < 0.003). Mice of the Ma stock also had a significantly longer maximal longevity than DC mice (9%, P = 0.04). The longest-lived Id mouse died at the age of 1450 days, which appears to exceed the previous longevity record for fully fed, non-mutant mice. The life table of the Po mice resembled that of the DC controls. Ma and Id mice differ from DC mice in several respects: both are shorter and lighter, and females of both stocks, particularly Id, are much slower to reach sexual maturity. As young adults, Id mice have lower levels of insulin-like growth factor 1 (IGF-I), leptin, and glycosylated hemoglobin compared with DC controls, implicating several biochemical pathways as potential longevity mediators. The results support the idea that inadvertent selection for rapid maturation and large body size during the adaptation of the common stocks of laboratory mice may have forced the loss of natural alleles that retard the aging process. Genes present in the Id and Ma stocks may be valuable tools for the analysis of the physiology and biochemistry of aging in mice.


Metastasis-Associated 1 (MTA1) Gene Expression Promotes Angiogenesis in Mouse Xenografts from Human Non-Small Cell Lung Cancer (NSCLC) Cells.
BACKGROUND This study aimed to investigate the effects of metastasis-associated 1 (MTA1) gene expression and gene silencing in human non-small cell lung cancer (NSCLC) cells in vitro and on angiogenesis in tumor xenografts in vivo in nude mice. MATERIAL AND METHODS Human H460 and H1299 NSCLC cell lines underwent transfection with lentiviral transfer plasmids (lenti) and short-interfering RNA (si-RNA) and included a control group, a lenti-MTA1 group, a lenti-si-MTA1 group, a lenti control group, and a si-RNA control group. Quantitative reverse transcription polymerase chain reaction (qRT-PCR) was used to detect MTA1 gene expression after cell transfection. MTA1 transfection was more effective in H460 cells, which were selected for further in vivo studies. Sixty Balb/c nude mice, containing human H460 cell tumor xenografts, included a control group (N=20), a lenti-MTA1 group (N=20), and a lenti-si-MTA1 group (N=20). Tumor tissue immunohistochemistry was used to detect the expression of MTA1 protein and microvessel density (MVD) using CD31. Western blot was used to quantify the expression of cyclooxygenase-2 (COX-2), angiopoietin 1/2 (Ang1/2), hypoxia-inducible factor 1-a (HIF-1a), and vascular endothelial growth factor (VEGF). RESULTS MTA1 silencing with si-RNA significantly reduced the tumor growth rate in nude mice (p<0.01), reduced tumor MVD, and 70% of mice survived for more than 30 days. MTA1 overexpression resulted in the death of all mice at 30 days after tumor inoculation and upregulated the expression of COX-2, Ang1/2, HIF-1a and VEGF, which were down-regulated by MTA1 silencing. CONCLUSIONS MTA1 gene expression promoted angiogenesis in mouse xenografts from human NSCLC cells.


[Fear of dental treatment among children. Influence of experience and psychological functioning].
This research considers whether fear of dental treatment among children is linked with a prior (negative) experience with local anaesthesia or with psychological functioning. Furthermore, the study investigated whether these factors play a role in anxious behaviour before and during a local anaesthetic injection. In this study 128 children between the ages of 4 and 11 took part. With the help of video recordings, the anxious behaviour before and during the local anaesthetic injection was scored using the Venham-scale. During the treatment one of the child's parents or guardians was asked if the child had previously been given a local anaesthetic injection (experience). The fear of dental treatment and the child's psychological functioning were measured using the parents' version of the 'Dental Subscale' of the 'Children's Fear Survey Schedule' and the 'Strengths and Difficulties Questionnaire' respectively. Results showed a correlation between on one hand fear of dental treatment and on the other hand anxious behaviour before and during a local anaesthetic injection and psychological functioning. Furthermore, results indicated that children with fear of dental treatment exhibited more fearful behaviour before and during an injection with local anaesthetic. Only in the case of children whose psychological functioning was reduced there was a significant relationship between experience with local anaesthesia and fearful behaviour prior to the injection of a local anaesthetic.


Community-Engaged Bidirectional Crisis and Emergency Risk Communication With Immigrant and Refugee Populations During the COVID-19 Pandemic.
This study was conducted to assess an intervention that was created by a community-academic partnership to address COVID-19 health inequities. We evaluated a community-engaged bidirectional pandemic crisis and emergency risk communication (CERC) framework with immigrant and refugee populations during the COVID-19 pandemic. A 17-year community-engaged research partnership adopted a CERC framework in March 2020 to address COVID-19 prevention, testing, and socioeconomic impacts with immigrant and refugee groups in southeast Minnesota. The partnership used bidirectional communication between communication leaders and their social networks to refine messages, leverage resources, and advise policy makers. We conducted a mixed-methods evaluation for intervention acceptability, feasibility, reach, adaptation, and sustainability through multisource data, including email communications, work group notes, semistructured interviews, and focus groups. The intervention reached at least 39 000 people in 9 months. It was implemented as intended and perceived efficacy was high. Frequent communication between community and academic partners allowed the team to respond rapidly to concerns and facilitated connection of community members to resources. Framework implementation also led to systems and policy changes to meet the needs of immigrant and refugee populations. Community-engaged CERC is feasible and sustainable and can reduce COVID-19 disparities through shared creation and dissemination of public health messages, enhanced connection to existing resources, and incorporation of community perspectives in regional pandemic mitigation policies.


IL-1B and IL-1RN gene polymorphisms in rheumatoid arthritis: relationship with protein plasma levels and response to therapy.
To analyze the association of interleukin (IL-1) gene polymorphisms with susceptibility to, and severity of, rheumatoid arthritis (RA) patients, comparing them with the genotype distribution in healthy controls. Also, to assess the influence of IL1-B and IL1RN gene polymorphism on IL-1beta/IL-1Ra plasma levels and response to therapy. We tested the allelic distribution of IL-1B (-511 and +3953) and IL-1RN (variable number of tandem repeats) gene polymorphism in 126 RA patients and 178 healthy blood donors (HBDs). The patients were categorized into two subgroups in relation to the response to methotrexate (MTX) therapy. Group A included 70 RA patients in stable partial remission after 6 months of MTX treatment (MTX-R). Group B included 56 RA patients with active disease despite MTX therapy. This group received antitumor necrosis factor (TNF) biological drugs and were defined MTX-nonresponders (MTX-NR). None of the two IL-1B (-511 and +3953) gene polymorphisms were significantly different in frequency between RA patients and healthy controls. We observed an increased frequency of the rare allele IL1RN*3 in RA patients with active disease, not responding to MTX therapy (MTX-NR) (4.5%) vs MTX-R (3.6%) and healthy controls (0.8%). Interestingly, RA patients whose genotypes included the IL1RN*long allele (haplotype long-C-T) showed the worse response to MTX. HBDs harboring the IL1RN*2/2 genotype showed significantly lower levels of plasma IL1-Ra, but comparable levels of IL-1beta with regard to subjects with the presence of the IL1RN* long allele. Furthermore, the presence of the TT IL-1B +3953 genotype was associated with lower plasma levels of IL1-Ra, both in HBDs and in RA patients. Carriers of the IL1RN*2 allele responded better to infliximab therapy. The results of this study provide evidence of an association between the IL1RN*long allele and RA, the strongest association being observed in RA patients with an aggressive disease resistant to MTX treatment.


Effect of heatwaves and greenness on mortality among Chinese older adults.
Heatwaves and greenness have been shown to affect health, but the evidence on their joint effects is limited. We aim to assess the associations of the combined exposure to greenness and heatwaves. We utilized five waves (February 2000-October 2014) of the Chinese Longitudinal Healthy Longevity Survey (CLHLS), a prospective cohort of older adults aged 65. We defined heatwaves as the daily maximum temperature ≥92.5th percentile with duration ≥3 days. We calculated the number of heatwave days in one year before death to and cumulative Normalized Difference Vegetation Index (NDVI) during follow-up to assess individual long-term exposure to heatwaves and greenness. Cox proportional hazards models were used to assess the effects of greenness, heatwaves, and their interaction on mortality, adjusted for covariates. We conducted subgroup analyses by residence, gender, and age. There were 20,758 participants in our study, totaling 67,312 person-years of follow-up. The mean NDVI was 0·41 (SD 0.13), and the mean number of heatwave days was 8.92 (2.04). In the adjusted model, the mortality hazard ratio (HR) for each 3-day increase in heatwave days was 1.04 (95% CI 1.04, 1.05), each 0.1-unit decrease in cumulative NDVI was 1.06 (1.05, 1.07). In the adjusted model with an interaction term, the HR for the interaction term was 1.01 (1.01, 1.02) with a p-value less than 0.001. In our subgroup analyses, the HR for each 3-day increase in heatwave days was higher in urban areas than in rural areas (1.06 vs. 1.03), and the HR for 0.1-unit decrease in NDVI was higher in urban areas than in rural areas (1.08 vs. 1.04). Greenness can protect against the effect of heatwaves on mortality, and heatwaves affect the health effects of greenness. Urban dwellers have a higher response to the detrimental effect of heatwaves and a higher marginal benefit from greenness exposure.


[The development OF THE vestibular apparatus under conditions of weightlessness].
The spawn of the aquarium fish Brachydanio rerio was developing during 5--6 days under conditions of weightlessness (first on board the spaceship "Sojuz-16", then in the space station "Salut-4") in special aquariums "EMKON", in thermostable installations. Electron microscopically the embryos were found to have a well developed labyrinth in early developmental histologically and cytologically differentiated receptory structures of the macula utriculi and macula saccili. In contrast to controls, the experimental animals showed certain alterations in the otolite organization. In similar experiments the embryos of clawed frog Xenopus laevis in the stage of the tail bud were also placed in special containers "EMKON" and thermostable apparatus "Biotherm-4" and by the spaceship "Sojuz-17" were brought to the space station "Salut-4", where it stayed for 16 days. The initial embryos had already had a well developed acoustic vesicle with macula communis. Inspite of the preliminary load by start acceleration and staying under conditions of weightlessness, they reached the general development fairly similar to controls. As it was shown electron microscopically their labyrinth had highly histologically and cytologically differentiated structures. However, a disturbance of the development of the otolithic membrane and otoconia should be noted. The alterations observed in the otolithic membrane organization in experimental fishes and frogs may be explained by general disorders in calcium metabolism.


Efficacy and safety of esomeprazole compared with omeprazole in GERD patients with erosive esophagitis: a randomized controlled trial.
In patients with gastroesophageal reflux disease (GERD), esomeprazole, the S-isomer of omeprazole, has demonstrated pharmacological and clinical benefits beyond those seen with the racemic parent compound. This study was designed to further evaluate the efficacy and tolerability of esomeprazole relative to that of omeprazole in healing erosive esophagitis and resolving accompanying symptoms of GERD. Esomeprazole 40 mg was compared with omeprazole 20 mg once daily in 2425 patients with erosive esophagitis (Helicobacter pylori negative by serology) in an 8-wk, multicenter, randomized, double-blind, parallel-group study conducted in 163 centers throughout the US. The primary efficacy endpoint was the proportion of patients with healed esophagitis at wk 8. Secondary endpoints were the proportion of patients healed at wk 4, resolution of heartburn at wk 4, time to first resolution and sustained resolution of heartburn, and proportion of heartburn-free days and nights. Safety and tolerability were also assessed. Significantly more patients were healed with esomeprazole versus omeprazole at wk 8 (93.7% vs 84.2%, p < 0.001; life table estimates, intention-to-treat analysis). Healing rates at wk 4 were 81.7% and 68.7%, respectively. Esomeprazole was superior to omeprazole for all secondary measures and had a similar safety profile. The most common adverse events in both treatment groups were headache, diarrhea, and nausea. Esomeprazole demonstrates significantly greater efficacy than omeprazole in the treatment of GERD patients with erosive esophagitis. The tolerability and safety of esomeprazole are comparable to that of omeprazole. (Am


Predominant allylic hydroxylation at carbons 6 and 7 of 4 and 5-ene functionalized steroids by the thermophilic fungus Rhizomucor tauricus IMI23312.
This paper demonstrates for the first time transformation of a series of steroids (progesterone, androst-4-en-3,17-dione, testosterone, pregnenolone and dehydroepiandrosterone) by the thermophilic fungus Rhizomucor tauricus. All transformations were found to be oxidative (monohydroxylation and dihydroxylation) with allylic hydroxylation the predominant route of attack functionalizing the steroidal skeleta. Timed experiments demonstrated that dihydroxylation of progesterone, androst-4-en-3,17-dione and pregnenolone all initiated with hydroxylation on ring-B followed by attack on ring-C. Similar patterns of steroidal transformation to those observed with R. tauricus have been observed with some species of thermophilic Bacilli and mesophilic fungi. All metabolites were isolated by column chromatography and were identified by (1)H, (13)C NMR, DEPT analysis and other spectroscopic data. The application of thermophilic fungi to steroid transformation may represent a potentially rich source for the generation of new steroidal compounds as well as for uncovering inter and intraspecies similarities and differences in steroid metabolism.


Biochemical and pharmacokinetic effects of leucovorin after high-dose methotrexate in a murine leukemia model.
The administration of calcium leucovorin, either concurrently or after high dosages of methotrexate in L1210 leukemic mice, has both pharmacokinetic and biochemical effects in tumor cells and drug-limiting proliferative normal tissue in small intestine. A reduction in the maximal level of accumulation and retention of exchangeable drug (unbound to dihydrofolate reductase) in tissue could be demonstrated when calcium leucovorin was given simultaneously with methotrexate at equal or greater dosages than the latter. The dose dependence for calcium leucovorin-introduced drug loss is similar in both tissues and showed the expected variation when the time interval between methotrexate and calcium leucovorin doses was increased. With 400 mg methotrexate per kg, greater than 96 mg calcium leucovorin per kg were required maximally to affect overall drug retention in tissue 2 hr after drug, whereas only 24 mg calcium leucovorin per kg were required 16 hr after drug. Calcium leucovorin, given after methotrexate, induced synchronous recovery of DNA synthesis (measured by labeled deoxyruridine incorporation) in both small intestine and L1210 cells. An initial cycle of synthesis was induced in the presence of exchangeable levels of drug. Two hr after methotrexate, 12 mg/kg, calcium leucovorin induced an immediate but only partial (20 to 25% of control rate) recovery of synthesis with dose dependence from 3 to 12 mg calcium leucovorin per kg. Less synthesis was induced after 96 mg/kg and almost none after methotrexate, 400 mg/kg. With calcium leucovorin, 24 mg/kg, given 2 hr after methotrexate, 12 or 96 mg/kg, a major cycle of synthesis occurred when total drug levels approached the equivalence of the dihydrofolate reductase content. The magnitude of this cycle of synthesis in both L1210 cells and small intestine was the same as that seen in animals recovering from methotrexate alone. However, this is based on the assumption that an approximately equivalent relationship between DNA synthesis and labeled doexyuridine incorporation in each tissue during the period of maximal incorporation within the cycle. The major effect of calcium leucovorin, then, was to induce an earlier resumption of DNA synthesis as a consequence of the pharmacokinetic effect in each tissue. With calcium leucovorin, 24 or 400 mg/kg, given 16 hr after methotrexate, an identical effect on drug retention was observed in both L1210 cells and small intestine. Although there was a difference in the time course for recovery in small intestine at each dosage of calcium leucovorin, the recovery of DNA synthesis as drug levels approached the dihydrofolate reductase content was similar in magnitude. In L1210 cells, however, substantial recovery of synthesis to a comparable level and with a similar time-course occurred only after leucovorin, 400 mg/kg. Little or no recovery of DNA synthesis was observed after calcium leucovorin, 24 mg/kg, during the same time period. This dosage schedule (methotrexate, 400 mg/kg, s.c...


Effects of allochthonous dissolved organic matter input on microbial composition and nitrogen-cycling genes at two contrasting estuarine sites.
Heterotrophic bacteria are important drivers of nitrogen (N) cycling and the processing of dissolved organic matter (DOM). Projected increases in precipitation will potentially cause increased loads of riverine DOM to the Baltic Sea and likely affect the composition and function of bacterioplankton communities. To investigate this, the effects of riverine DOM from two different catchment areas (agricultural and forest) on natural bacterioplankton assemblages from two contrasting sites in the Baltic Sea were examined. Two microcosm experiments were carried out, where the community composition (16S rRNA gene sequencing), the composition of a suite of N-cycling genes (metagenomics) and the abundance and transcription of ammonia monooxygenase (amoA) genes involved in nitrification (quantitative PCR) were investigated. The river water treatments evoked a significant response in bacterial growth, but the effects on overall community composition and the representation of N-cycling genes were limited. Instead, treatment effects were reflected in the prevalence of specific taxonomic families, specific N-related functions and in the transcription of amoA genes. The study suggests that bacterioplankton responses to changes in the DOM pool are constrained to part of the bacterial community, whereas most taxa remain relatively unaffected.


Threshold Graph Limits and Random Threshold Graphs.
We study the limit theory of large threshold graphs and apply this to a variety of models for random threshold graphs. The results give a nice set of examples for the emerging theory of graph limits.