Seasonal and sex-specific changes in the gastrointestinal tracts of Peromyscus maniculatus.
Functional traits are phenotypic characteristics that contribute to fitness of individuals in dynamic and changing environments. In mammals, both categorical and continuous (e.g., quantitative) functional traits have been extensively utilized as proxies for diet, locomotion, and other aspects of species ecology, but there has been less focus on form and function of soft tissues. This is particularly true for the digestive system, which varies in size and complexity across Class Mammalia and plays a major role in the energetics of species. To guide more effective utilization of gastrointestinal (GI) morphology as a functional proxy in small mammal ecology, we examined how GI tracts (lengths and masses of four GI sections) varied within a population of deer mice (Peromyscus maniculatus) in the Southern Appalachian Mountains of North Carolina, United States. We collected samples of adult P. maniculatus monthly for 1 year and measured GI tracts to quantify variation with respect to seasonality and trophic level, providing insight into plasticity in this soft tissue trait over time. We found that season had a significant effect on the total length and wet mass of the GI tract, with January mice having the longest GI tracts and lengths being shortest in the summer. The relative shortening of the GI tract in summer corresponded with a partial trophic increase detected by stable isotope signatures. GI length and wet mass also were affected by reproduction, but males and females responded in sex-specific ways to demands of reproduction, with reproductively active males having shorter and lighter GI tracts than nonreproductively active males. Our study provides proof-of-concept for understanding population-level plasticity in a rarely collected soft tissue trait, which may also be complementary to standard craniodental measurements as a functional dietary proxy to understand mammalian ecology and community assembly.


Motif-based construction of a functional map for mammalian olfactory receptors.
We applied an automatic and unsupervised system to a nearly complete database of mammalian odor receptor genes. The generated motifs and gene classification were subjected to extensive and systematic downstream analysis to obtain biological insights. Two major results from this analysis were: (1) a map of sequence motifs that may correlate with function and (2) the corresponding receptor classes in which members of each class are likely to share specific functions. We have discovered motifs that have been implicated in structural integrity and posttranslational modification, as well as motifs very likely to be directly involved in ligand binding. We further propose a combinatorial molecular hypothesis, based on unique combinations of the observed motifs, that provides a foundation for understanding the generation of a large number of ligand binding sites.


Serum uPAR as Biomarker in Breast Cancer Recurrence: A Mathematical Model.
There are currently over 2.5 million breast cancer survivors in the United States and, according to the American Cancer Society, 10 to 20 percent of these women will develop recurrent breast cancer. Early detection of recurrence can avoid unnecessary radical treatment. However, self-examination or mammography screening may not discover a recurring cancer if the number of surviving cancer cells is small, while biopsy is too invasive and cannot be frequently repeated. It is therefore important to identify non-invasive biomarkers that can detect early recurrence. The present paper develops a mathematical model of cancer recurrence. The model, based on a system of partial differential equations, focuses on tissue biomarkers that include the plasminogen system. Among them, only uPAR is known to have significant correlation to its concentration in serum and could therefore be a good candidate for serum biomarker. The model includes uPAR and other associated cytokines and cells. It is assumed that the residual cancer cells that survived primary cancer therapy are concentrated in the same location within a region with a very small diameter. Model simulations establish a quantitative relation between the diameter of the growing cancer and the total uPAR mass in the cancer. This relation is used to identify uPAR as a potential serum biomarker for breast cancer recurrence.


Use of intramuscular ziprasidone for the control of acute psychosis or agitation in an inpatient geriatric population: an open-label study.
Objective. This open-label study examined the safety and efficacy of ziprasidone intramuscular with geriatric patients experiencing psychosis or agitation.Design. During an inpatient stay, consenting subjects who became acutely psychotic or agitated received ziprasidone intramuscular 10mg q 6 to 8 hours, up to a maximum dose of 20mg/24 hours. A within-group repeated measures design was employed to study whether the use of ziprasidone over the 24-hour observation period contributed to decreased agitation or to extrapyramidal side effects. The data were analyzed using an Analysis of Variance with trend analysis.Setting. The study was conducted on the geriatric psychiatry inpatient unit at the University of Toledo Medical Center, Toledo, Ohio.Participants. Fourteen patients, six men and eight women with mean age 77+/-8 years, participated in this study. Each patient had a diagnosis of dementia, co-occurring with one of the following: delirium, major depressive disorder with psychotic features, schizophrenia, bipolar disorder, or schizoaffective disorder.Measurements. The Brief Psychiatric Rating Scale, Delirium Rating Scale, and the Behavioral Activity Rating Scale were obtained at baseline and at 0.5, 2, and 24 hours after the first dose of ziprasidone intramuscular.Results. Overall, physiologic measures that would indicate undesirable side effects, including QTc intervals, remained unchanged pre- and post-study. However, there were significant improvements in scores on a variety of measures assessing agitation or psychosis.Conclusion. This study suggests that ziprasidone intramuscular may be a safe and effective short-term treatment for agitated or psychotic geriatric patients, and, therefore, additional studies should be conducted to confirm these findings.


Cross-cultural validation and psychometric evaluation of the Participation and Environment Measure for Children and Youth in Korea.
To develop the Korean version of the Participation and Environment Measure for Children and Youth (KPEM-CY) and examine its psychometric properties. The PEM-CY was cross-culturally translated into Korean using a specific guideline: pre-review of participation items, forward/backward translation, expert committee review, pre-test of the KPEM-CY and final review. To establish internal consistency, test-retest reliability and construct validity of the KPEM-CY, 80 parents of children with disabilities aged 5-13 years were recruited in South Korea. Across the home, school and community settings, 76% of participation items and 29% of environment items were revised to improve their fit with Korean culture. Internal consistency was moderate to excellent (0.67-0.92) for different summary scores. Test-retest reliability was excellent (>0.75) in the summary scores of participation frequency and extent of involvement across the three settings and moderate to excellent (0.53-0.95) in all summary scores at home. Child's age, type of school and annual income were the factors that significantly influenced specific dimensions of participation and environment across all settings. Results indicated that the KPEM-CY is equivalent to the original PEM-CY and has initial evidence of reliability and validity for use with Korean children with disabilities. Implications for rehabilitation Because 'participation' is a key outcome of the rehabilitation, measuring comprehensive participation of children with disabilities is necessary. The PEM-CY is a parent-report survey measure to assess comprehensive participation of children and youth and environment, which affect their participation, at home, school and in the community. A cross-cultural adaptation process is mandatory to adapt the measurement tool to a new culture or country. The Korean PEM-CY has both reliability and validity and can therefore generate useful clinical data for Korean children with disabilities.


Cervical Spine Evaluation in Pediatric Trauma: A Review and an Update of Current Concepts.
The clinical presentation and diagnostic workup in pediatric cervical spine injuries (CSI) are different from adults owing to the unique anatomy and relative immaturity. The current article reviews the existing literature regarding the uniqueness of these injuries and discusses the current guidelines of radiological evaluation. A PubMed search was conducted using keywords "paediatric cervical spine injuries" or "paediatric cervical spine trauma." Six hundred and ninety two articles were available in total. Three hundred and forty three articles were considered for the review after eliminating unrelated and duplicate articles. Further screening was performed and 67 articles (original articles and review articles only) related to pediatric CSI were finally included. All articles were reviewed for details regarding epidemiology, injury patterns, anatomic considerations, clinical, and radiological evaluation protocols. CSIs are the most common level (60%-80%) for pediatric Spinal Injuries (SI). Children suffer from atlantoaxial injuries 2.5 times more often than adults. Children's unique anatomical features (large head size and highly flexible spine) predispose them to such a peculiar presentation. The role of National Emergency X-Ray Utilization Study, United State (NEXUS) and Canadian Cervical Spine Rule criteria in excluding pediatric cervical injury is questionable but cannot be ruled out completely. The minimum radiological examination includes 2- or 3-view cervical X-rays (anteroposterior, lateral ± open-mouth odontoid views). Additional radiological evaluations, including computerized tomography (CT) and magnetic resonance imaging (MRI) are obtained in situations of abnormal physical examination, abnormal X-rays, inability to obtain adequate X-rays, or to assess cord/soft-tissue status. The clinical criteria for cervical spine injury clearance can generally be applied to children older than 2 years of age. Nevertheless, adequate caution should be exercised before applying these rules in younger children. Initial radiographic investigation should be always adequate plain radiographs of cervical spine. CT and MRI scans should only be performed in an appropriate group of pediatric patients.


Creating institutional flexibility for adaptive water management: insights from two management agencies.
Adaptive management is an experimental approach used by water management agencies around the world to manage and restore aquatic ecosystems. The effectiveness of the approach can often be constrained by inflexible institutional arrangements. In this paper we compare two cases where agencies have implemented adaptive management to manage and restore aquatic ecosystems. Our aim was to understand practitioners' perceptions of how institutional flexibility can be created for adaptive management. We interviewed 14 adaptive management practitioners working in the Murray-Darling Basin, Australia and 14 practitioners in Southern Florida, United States of America. We found that in both cases, just enough flexibility was created to enable experimentation, but informal institutional arrangements tended to constrain adaptive management. We also found that adaptive management was effective when an agency adopted collaborative and distributed leadership, but these leadership styles were difficult to sustain, and not always appropriate when attempting to create institutional flexibility. Our results illustrate how agencies, stakeholders and researchers can develop a shared understanding of how to manage and restore aquatic ecosystems, which in turn, helps create institutional flexibility for an agency to manage adaptively.


Improved survival after pneumococcus in splenectomized and nonsplenectomized mice with Corynebacterium parvum.
Splenectomy increases the susceptibility to infections with certain bacteria, particularly Streptococcus pneumoniae. Because the immunomodulator Corynebacterium parvum expands the phagocytic cell compartment and enhances reticuloendothelial function, we tested the effect of C parvum in mice challenged with aerosolized pneumococci. Mice splenectomized seven days before pneumococcal challenge and treated intraperitoneally with 700 micrograms of C parvum immediately after exposure were protected when compared with splenectomized or sham-operated saline-injected controls. Analysis of proportional hazards showed the risk of dying in order of greatest to least as follows: splenectomy/saline, sham/saline, splenectomy/C parvum and sham/C parvum. The benefits of an intact spleen and C parvum seemed to be additive in their protective effects after aerosol pneumococcal challenge. After intravenous challenge, bloodstream clearance was improved in sham-operated mice at three days after C parvum injection compared with saline-injected sham-operated controls and C parvum-injected splenectomized mice. A significant improvement in bacterial clearance did not occur until seven days after C parvum treatment in splenectomized mice. The results demonstrate the value of a nonspecific immunomodulator for enhancing the defense mechanisms of both normal and splenectomized animals.


Exposure to cutting oils and its relation to skin tumors and premalignant skin lesions on the hands and forearms.
In a cohort study the incidence of skin tumors on the hands and forearms of 682 turners exposed to cutting oils was compared to the incidence of the general male population and of office workers. Among the 682 turners, five premalignant squamous cell tumors and eight keratoacanthomas were found between 1960 and 1980. Five of this total of 13 turners with primary skin tumors had another skin tumor as well. Among the 682 men there were four cases of scrotal cancer. Among the 375 office workers examined, no malignant or premalignant tumors or keratoacanthomas were found on the hands or forearms. No primary skin tumors were found among the turners after 1975, when the acid-refined mineral oils were replaced by solvent-refined oils containing a far lower concentration of polyaromatic hydrocarbons, which are probably the cancerogenic agent in the oils. The study shows a greatly increased risk of skin tumors on the hands and forearms of workers exposed to acid-refined mineral oils. Individual susceptibility also seems to be indicated.


The MADS-domain transcriptional regulator AGAMOUS-LIKE15 promotes somatic embryo development in Arabidopsis and soybean.
The MADS-domain transcriptional regulator AGAMOUS-LIKE15 (AGL15) has been reported to enhance somatic embryo development when constitutively expressed. Here we report that loss-of-function mutants of AGL15, alone or when combined with a loss-of-function mutant of a closely related family member, AGL18, show decreased ability to produce somatic embryos. If constitutive expression of orthologs of AGL15 is able to enhance somatic embryo development in other species, thereby facilitating recovery of transgenic plants, then AGL15 may provide a valuable tool for crop improvement. To test this idea in soybean (Glycine max), a full-length cDNA encoding a putative ortholog of AGL15 was isolated from soybean somatic embryos. Subsequently, the corresponding genomic region of the gene was obtained. This gene, designated GmAGL15, encodes a protein with highest similarity to AGL15 from Arabidopsis (Arabidopsis thaliana) and Brassica napus that accumulates to its highest amount in embryos in these species. Like Arabidopsis and Brassica AGL15, GmAGL15 was preferentially expressed in developing embryos. When ectopically overexpressed the soybean protein was able to enhance somatic embryo development in soybean.


Frequent use of emergency departments in Saudi public hospitals: implications for primary health care services.
This study aims to determine the prevalence, factors and reasons associated with the frequent use of public emergency departments (EDs) in Riyadh, Saudi Arabia. The study employed a self-administered questionnaire to collect data from adult patients on aspects such as demographic characteristics, accessibility, and health-related factors. Bivariate and multivariate analyses were used to explore the issue. The results indicated that about 30% of respondents were considered as "frequent users" of the EDs. Males, older patients, those living closer to EDs, and patients who had chronic illnesses or were hospitalized in the past year were more likely to use EDs frequently. Moreover, the study found that patients who were labeled as "frequent users" of the EDs were also frequent users of "other" health care facilities. Health decision makers should facilitate primary health centers with necessary resources that fulfill the patients' health needs and reduce the burden on EDs.


Automatic Target Detection from Satellite Imagery Using Machine Learning.
Object detection is a vital step in satellite imagery-based computer vision applications such as precision agriculture, urban planning and defense applications. In satellite imagery, object detection is a very complicated task due to various reasons including low pixel resolution of objects and detection of small objects in the large scale (a single satellite image taken by Digital Globe comprises over 240 million pixels) satellite images. Object detection in satellite images has many challenges such as class variations, multiple objects pose, high variance in object size, illumination and a dense background. This study aims to compare the performance of existing deep learning algorithms for object detection in satellite imagery. We created the dataset of satellite imagery to perform object detection using convolutional neural network-based frameworks such as faster RCNN (faster region-based convolutional neural network), YOLO (you only look once), SSD (single-shot detector) and SIMRDWN (satellite imagery multiscale rapid detection with windowed networks). In addition to that, we also performed an analysis of these approaches in terms of accuracy and speed using the developed dataset of satellite imagery. The results showed that SIMRDWN has an accuracy of 97% on high-resolution images, while Faster RCNN has an accuracy of 95.31% on the standard resolution (1000 × 600). YOLOv3 has an accuracy of 94.20% on standard resolution (416 × 416) while on the other hand SSD has an accuracy of 84.61% on standard resolution (300 × 300). When it comes to speed and efficiency, YOLO is the obvious leader. In real-time surveillance, SIMRDWN fails. When YOLO takes 170 to 190 milliseconds to perform a task, SIMRDWN takes 5 to 103 milliseconds.


[N-terminal fragment of the brain natriuretic peptide and proinflammatory cytokines in patients with ischemic heart disease].
To determine plasmic concentrations of NT-proBNP, TNFalpha and IL-6 in patients with coronary heart disease (CHD) complicated with chronic cardiac failure (CCF); to compare these parameters with hemodynamic and functional ones. A total of 157 CHD patients (84.1% males, mean age 54.3 +/- 6.8 years) were divided into 3 groups matched by sex and age according to the presence of CCF and left ventricular (LV) or systolic dysfunction. CCF patients with LV dysfunction or intact LV function had much higher plasmic concentrations of neurohumoral mediators than CHD patients without CCF. A plasmic NT-proBNP level over 1000 fmol/ml was associated with a 5.5-fold increase in the relative risk of LV diastolic dysfunction for CCF patients (rR 5.6; 95% CI 1.4-30.0; p = 0.0065) while an IL-6 level over 6.1 pg/ml was associated with a 9-fold increase in this risk (rR 8.9; 95% CI 2.3-35.4; p = 0.00001). Plasmic NT-proBNP and IL-6 levels went up in correlation with a CCF functional class. A plasmic level of TNFalpha correlated with anginal functional class. Changes in plasmic levels of the above neuromediators are associated with changes in some LV function and functional parameters of CCF patients.


Flow regulation and fragmentation imperil pelagic-spawning riverine fishes.
Flow regulation and fragmentation of the world's rivers threaten the integrity of freshwater ecosystems and have resulted in the loss or decline of numerous fish species. Pelagic-spawning fishes (pelagophils) are thought to be particularly susceptible to river regulation because their early life stages (ichthyoplankton) drift until becoming free-swimming, although the extent of transport is largely unknown. Transport velocity and distance were determined for passively drifting particles, which mimicked physical properties of ichthyoplankton, in two large, regulated rivers (Rio Grande and Pecos River) of the arid Southwest United States. Particle drift data were incorporated into celerity-discharge equations (r2 > 0.90; P < 0.001), and reach-specific transport velocity was modeled as a function of discharge. Transport velocities of particles exceeded 0.7 m/s in all river reaches during typical spawning flows (i.e., reservoir releases or rainstorms) and were greatest in highly incised and narrow channel reaches. Mean transport distance of particles released in the Pecos River during sustained reservoir flows (141.1 km; 95% CI = 117.0-177.5 km) was significantly longer than during declining reservoir flows that mimicked a natural rainstorm (52.4 km; 95% CI = 48.8-56.5 km). Mean transport distance of particles in the Rio Grande during sustained reservoir flows was 138.7 km (95% CI = 131.0-147.2 km). There are 68 dams and 13 reservoirs that fragment habitats and regulate flow in the Rio Grande Basin (Rio Grande and Pecos River) in areas historically occupied by pelagophils. While the basin historically provided 4029 km of free-flowing riverine habitat, reservoir habitat now represents > 10% of the longitudinal distance. Only five unfragmented nonreservoir reaches > 100 km remain in the Rio Grande, and two remain in the Pecos River. Pelagophils were extirpated from all reservoirs and from nearly all short, fragmented reaches (< 100 km) of the Rio Grande Basin, but at least some fraction persisted in all longer reaches (> 100 km). The recovery and long-term persistence of pelagophils in regulated rivers, including those in this study, will likely depend on reestablishment and protection of long unfragmented reaches coupled with mimicry of the natural flow regime.


Role of ovarian hormones on hoarding in rats--feedback mechanism.
The adult female rats hoard large quantities of food during proestrus and continue hoarding if ovariectomised at proestrus (Group I). The hoarding score is lowest in diestrus and continue to remain low if ovariectomised during this phase (Group II). The scores in Group I and Group II are reversed in intracerebral administration of Progesterone and Estradiol Benzoate respectively into preoptic areas. The significance of the response is discussed on the basis of built-in feedback mechanism for hoarding cyclicity.


Combined Assessment of Serum Alpha-Synuclein and Rab35 is a Better Biomarker for Parkinson's Disease.
It is essential to develop a reliable predictive serum biomarker for Parkinson's disease (PD). The accumulation of alpha-synuclein (αSyn) and up-regulated expression of Rab35 participate in the etiology of PD. The purpose of this investigation was to determine whether the combined assessment of serum αSyn and Rab35 is a useful predictive biomarker for PD. Serum levels of αSyn or Rab35 were determined in serum samples from 59 sporadic PD patients, 19 progressive supranuclear palsy (PSP) patients, 20 multiple system atrophy (MSA) patients, and 60 normal controls (NC). Receiver operating characteristics (ROC) curves were calculated to determine the diagnostic accuracy of αSyn or/and Rab35 in discriminating PD patients from NC or atypical parkinsonian patients. The levels of αSyn and Rab35 were increased in PD patients. The serum level of Rab35 was positively correlated with that of αSyn in PD patients. Compared to analyzing αSyn or Rab35 alone, the combined analysis of αSyn and Rab35 produced a larger area under the ROC curve and performed better in discriminating PD patients from NC, MSA patients, or PSP patients. When age was dichotomized at 55, 60, 65, or 70 years, the combined assessment of αSyn and Rab35 for classifying PD was better in the group below the cutoff age than in the group above the cutoff age. Combined assessment of serum αSyn and Rab35 is a better biomarker for discriminating PD patients from NC or atypical parkinsonian patients, and is a useful predictive biomarker for younger sporadic PD patients.


Sequence of the 3'-terminal region of turnip mosaic virus RNA and the capsid protein gene.
A sequence of 1801 nucleotides originating from the 3' end region of turnip mosaic virus (TuMV) RNA was cloned using the polymerase chain reaction and found to contain one long open reading frame (ORF). The amino acid sequence of three different regions of the isolated TuMV capsid protein (including the NH2 terminus) was determined and these partial sequences were found in the translation product predicted to be encoded by the large ORF. The data suggested that the TuMV capsid protein was a product arising from the maturation of a larger polyprotein, as observed for other potyviruses. Furthermore, the putative cleavage site corresponded to a glutamine-alanine dipeptide, a site commonly used in plant virus polyprotein processing. The capsid protein cistron was composed of 864 nucleotides and corresponded to a region encoding 288 amino acids with a calculated Mr of 33,186; the adjacent 3' non-coding region was 667 nucleotides long. The deduced amino acid sequence of the TuMV capsid protein is closely related to other potyvirus capsid proteins, with most of the variation being found within the NH2-terminal region.


RELATIONSHIP BETWEEN VITAMIN D DEFICIENCY AND CHRONIC PERIODONTITIS.
Vitamin D deficiency may be associated with increased risk of chronic periodontitis. Aims - to clarify the relationship between vitamin D deficiency and Chronic periodontitis and to evaluate the effect of vitamin D on periodontal index. The investigation was carried out on 45 participants of ages within the range of (30-45 years) who were attending the private dental clinics. Diagnosis of chronic periodontitis was established depending on dental history, clinical examinations (periodontal indices). All participants were examined by the same dentist. They were classified into three groups: Group 1 (control negative): (15) participants with normal serum vitamin D3 level and with pocket depth ≤3 mm, good oral health and normal periodontal tissues and no previous history of periodontal diseases. Group 2 (control positive): (15) participants with normal serum vitamin D3 level and periodontitis with pocket depth ≥5 mm, they received placebo medication orally, Group3(treatment): (15) participants with vitamin D3 deficiency (below 30 IU), and periodontitis with pocket depth ≥5 mm, they received oral Vitamin D3 fast acting liquid soft gel capsule 2000 IU /day for 3 months. Serum Vitamin D level was measured before and after the study, 3 blood samples were taken from each participant at 0,45,90 days, for research examinations. The criteria of patients' selection include apparently looked healthy individuals, non-pregnant or lactating females. Vitamin D deficiency group (below 30 IU), there was no history of vitamin D allergy and did not take any medication or supplements or herbals for the last 1month, non-smoking, and non-alcoholic. Deep scaling and root planning were done for every participant in all groups (except control negative) to reach the base line for periodontal index. A written instruction was supplied to each patient about standard oral hygiene home care. After one week, the periodontal indices and radiographical examination was measured for all participants with blood collection and after 45,90 days. Vitamin D level measured before and after research steps. There was significant reduction in periodontal indices in 45, 90 days of the study which mean good response to the treatment and improvement in pocket depth. Conclusion: Vitamin D3 supplement can be a good adjuvant in chronic periodontitis.


A Multicenter Study To Evaluate the Performance of High-Throughput Sequencing for Virus Detection.
The capability of high-throughput sequencing (HTS) for detection of known and unknown viruses makes it a powerful tool for broad microbial investigations, such as evaluation of novel cell substrates that may be used for the development of new biological products. However, like any new assay, regulatory applications of HTS need method standardization. Therefore, our three laboratories initiated a study to evaluate performance of HTS for potential detection of viral adventitious agents by spiking model viruses in different cellular matrices to mimic putative materials for manufacturing of biologics. Four model viruses were selected based upon different physical and biochemical properties and commercial availability: human respiratory syncytial virus (RSV), Epstein-Barr virus (EBV), feline leukemia virus (FeLV), and human reovirus (REO). Additionally, porcine circovirus (PCV) was tested by one laboratory. Independent samples were prepared for HTS by spiking intact viruses or extracted viral nucleic acids, singly or mixed, into different HeLa cell matrices (resuspended whole cells, cell lysate, or total cellular RNA). Data were obtained using different sequencing platforms (Roche 454, Illumina HiSeq1500 or HiSeq2500). Bioinformatic analyses were performed independently by each laboratory using available tools, pipelines, and databases. The results showed that comparable virus detection was obtained in the three laboratories regardless of sample processing, library preparation, sequencing platform, and bioinformatic analysis: between 0.1 and 3 viral genome copies per cell were detected for all of the model viruses used. This study highlights the potential for using HTS for sensitive detection of adventitious viruses in complex biological samples containing cellular background. IMPORTANCE Recent high-throughput sequencing (HTS) investigations have resulted in unexpected discoveries of known and novel viruses in a variety of sample types, including research materials, clinical materials, and biological products. Therefore, HTS can be a powerful tool for supplementing current methods for demonstrating the absence of adventitious or unwanted viruses in biological products, particularly when using a new cell line. However, HTS is a complex technology with different platforms, which needs standardization for evaluation of biologics. This collaborative study was undertaken to investigate detection of different virus types using two different HTS platforms. The results of the independently performed studies demonstrated a similar sensitivity of virus detection, regardless of the different sample preparation and processing procedures and bioinformatic analyses done in the three laboratories. Comparable HTS detection of different virus types supports future development of reference virus materials for standardization and validation of different HTS platforms.


Serum alpha-fetoprotein predicts treatment outcome in chronic hepatitis C patients regardless of HCV genotype.
We examined the association between serum alpha-fetoprotein (AFP) level and sustained virological response (SVR) in 93 chronic hepatitis C patients. The SVR rate was much higher among patients with serum AFP levels below rather than above the median value (5.7 ng/ml) (58.7% and 19.2%, respectively; P<0.0001). Serum AFP should be added to the list of factors predictive of treatment response in chronic hepatitis C.


Assessment of coconut water added to Numida meleagris egg yolk as cryoprotectant for goat semen.
Effects were assessed of the dilutants TRIS and ACP - 101c® with the addition of different guinea fowl (Numida meleagris) egg yolk concentrations. Fifteen ejaculates were collected from five goats of the Anglo Nubian breed. The ejaculates were pooled and then divided into 12 groups, two control groups (GC1 TRIS, with 2.5% Gallus gallus domesticus hen egg yolk GOGD), (GC2 Control Group ACP - 101c®, with the addition of 2.5% Gallus gallus domesticus hen egg yolk GOGD) and ten experimental groups (EG), containing TRIS and ACP added with different concentrations of egg yolk from guinea hen (Numida meleagris) (TRIS 2,5% GONM; TRIS 5% GONM; TRIS 10% GONM; TRIS 15% GONM; TRIS 20% GONM; ACP® 2,5% GONM; ACP® 5% GONM; ACP® 10% GONM; ACP® 15% GONM; ACP® 20% GONM). Then cryopreservation was carried out and the samples stored in liquid nitrogen (-196 °C). After seven days, the samples were thawed and assessed for spermatic kinetics, immunofluorescence and sperm morphology. Analysis of GOMN by the CASA system showed that the various parameters were similar to those of GOGD (P>0.05). The membrane integrity, mitochondrial potential and the acrosome were not influenced by the treatment (P>0.05) nor by the dilutant used for cryopreservation (P>0.05). The spermatic morphology was also preserved by the different GOGD and GONM concentrations in the ACP® and TRIS dilutants, with no statistically significant differences (P<0.05). It was concluded that Numida meleagris egg yolk, as external membrane cryoproctant added to the dilutants ACP-101c® and TRIS, improved goat semen quality.


Magnetic resonance angiography in trauma.
Following blunt or penetrating trauma to the head and neck, a variety of traumatic vascular injuries may occur. Often the clinical presentation of a craniocervical arterial injury is delayed and neuroimaging studies are necessary to evaluate for delayed findings of intracranial infarction or hemorrhage. In this setting, magnetic resonance imaging (MRI) and magnetic resonance angiography (MRA) may allow a prompt noninvasive diagnosis of craniocervical vascular injury. MRA may be helpful in selecting those patients requiring conventional angiography and allows a noninvasive follow-up evaluation of arterial injury following institution of therapy.


Ultrasonic field modeling: a comparison of analytical, semi-analytical, and numerical techniques.
Modeling ultrasonic fields in front of a transducer in the presence and absence of a scatterer is a fundamental problem that has been attempted by different techniques: analytical, semi-analytical, and numerical. However, a comprehensive comparison study among these techniques is currently missing in the literature. The objective of this paper is to make this comparison for different ultrasonic field modeling problems with various degrees of difficulty. Four fundamental problems are considered: a flat circular transducer, a flat square transducer, a circular concave transducer, and a point focused transducer (concave lens) in the presence of a cavity. The ultrasonic field in front of a finite-sized transducer can be obtained by Huygens-Fresnel superposition principle that integrates the contributions of several point sources distributed on the transducer face. This integral which is also known as the Rayleigh integral or Rayleigh-Sommerfeld integral (RSI) can be evaluated analytically for obtaining the pressure field variation along the central axis of the transducer for simple geometries, such as a flat circular transducer. The semi-analytical solution is a newly developed mesh-free technique called the distributed point source method (DPSM). The numerical solution is obtained from finite element analysis. Note that the first three problems study the effect of the transducer size and shape, whereas the fourth problem computes the field in presence of a scatterer.


Formation of soy protein isolate-dextran conjugates by moderate Maillard reaction in macromolecular crowding conditions.
Several methods have been reported for the conjugation of proteins with polysaccharides. Protein-polysaccharide conjugates can be formed by traditional dry heating, but this process is not attractive from an industrial viewpoint, and no commercial conjugates have been manufactured in this way. In the present study, in order to develop a more practical reaction method, macromolecular crowding was used to attach polysaccharides to proteins. Soy protein isolate-dextran conjugates (SDCs) were prepared via the initial stage of the Maillard reaction in macromolecular crowding conditions. The impact of various processing conditions on the formation of SDCs was investigated. The optimal conditions chosen from the experiments were a soy protein isolate/dextran ratio of 1:1 (w/w), a pH of 6.5, a reaction temperature of 60 °C and a reaction time of 30 h. Circular dichroism spectroscopy showed that the secondary and tertiary structures of the conjugates were changed significantly. Structural flexibility increased, allowing better display of their functional characteristics. The conjugates had a composition with various sizes, especially macromolecules, according to gel permeation chromatography. Thermal analysis showed that the thermal stability of the conjugates was improved. The production of SDCs under macromolecular crowding conditions appears to be an effective and promising technique, representing an advance over classic protein glycosylation methods.


Barriers to physical activity and healthy diet among breast cancer survivors: A multilevel perspective.
Cancer survivors engage in suboptimal levels of health behaviours and report many barriers to health behaviours, but we lack a solid understanding of the different levels of barriers and how they relate to enacted health behaviours. To address these issues, we conducted mixed-method research in 97 breast cancer survivors. Participants' barriers to physical activity (PA) and healthy diet, asked as an open-ended question, were coded as individual-level, social-level, and organisational/environmental-level for each health behaviour. Moderate-to-vigorous PA and fruit and vegetable (F&V) intake were assessed. Most participants perceived at least one PA (72.7%) and diet (64.9%) individual-level barrier (e.g. physical symptoms/injury); only 15.2% (PA) and 15.6% (diet) reported at least one social-level barrier (e.g. family obligations). About 28.8% (PA) and 29.9% (diet) perceived at least one organisational/environmental-level barrier (e.g. job demand, cost of F&V). Survivors perceiving individual-level dietary barriers consumed less F&V (-.65 servings/day) than those not perceiving dietary barriers at this level. Survivors perceiving social-level dietary barriers reported marginally lower F&V intake (-.65 servings/day) than their counterparts. Those perceiving organisational/environmental-level PA barriers reported marginally fewer minutes (-44.30/week) of moderate-to-vigorous PA than their counterparts. Barriers at multiple levels should be addressed to improve health behaviours among breast cancer survivors.


Purification and properties of bovine caeruloplasmin.
A novel method is reported for isolation of bovine caeruloplasmin from plasma; it involves a rapid and mild procedure, namely two column chromatographies with stepwise elution and one (NH4)2SO4 precipitation, and results in a proteolytically undegraded homogeneous protein. The general structure of the protein, as evaluated by molecular-weight determination and amino acid composition, is very similar to that established for human and rat caeruloplasmin. Copper determination and e.p.r. spectral analysis on the native and NO-treated protein gave a metal-to-protein stoichiometry of six atoms of copper per molecule. Three copper atoms were detectable by e.p.r., with Type 2/Type 1 ratio = 1 : 3 in most samples. The protein is very sensitive to storage and/or handling. A component was isolated from aged samples, which was found to contain approximately four copper atoms per 125000 daltons, two of which were detectable by e.p.r. with the characters of Type 2 copper. However, the same component was found to be present, although to a lesser extent, in the fresh preparation and does not seem to be related to proteolytic degradation. This component has no oxidase activity. On the basis of these results it is suggested that caeruloplasmin molecules are intrinsically heterogeneous with respect to both copper content and copper type, and this can explain the intriguing stoichiometry regarding the different types of copper centres.


Role of the glucagon receptor COOH-terminal domain in glucagon-mediated signaling and receptor internalization.
The binding of glucagon to its hepatic receptor is known to result in a number of effects, including the intracellular accumulation of cAMP, the mobilization of intracellular Ca2+, and the endocytosis of glucagon and its receptor into intracellular vesicles. In this study, we begin to define the functional role of the COOH-terminal tail of the human glucagon receptor in glucagon-stimulated signal transduction and receptor internalization. We have created and expressed in Chinese hamster ovary (CHO) cells five truncation mutants in which the COOH-terminal 24, 56, 62, 67, and 73 amino acids have been removed. Cells expressing relevant truncated receptors were assayed for cell surface expression by immunofluorescence, for ligand-binding properties, for cAMP and Ca2+-mediated signal transduction properties, and for receptor endocytosis. In addition, a mutant receptor containing seven serine-to-alanine mutations in the COOH-terminal tail was studied. Our results reveal the following: 1) a region of the COOH-terminal tail that is required for proper cell surface expression, 2) the COOH-terminal 62 amino acids, which comprise the majority of the tail, are not required for ligand binding, cAMP accumulation, or Ca2+ mobilization, and 3) phosphorylation of the COOH-terminal tail is crucial for glucagon-stimulated receptor endocytosis.


Immunotherapy for gastric cancers: emerging role and future perspectives.
The broad use of immunotherapy is revolutionizing the treatment paradigms of many solid tumors. Although chemotherapy remains the treatment backbone for advanced gastric cancer, improvements in its molecular characterization and progresses in understanding its underpinning biology have supported clinical development of novel immunotherapies. However, the results of recent trials testing these new agents raise the question on how to identify the patients that could greatly benefit. Areas covered: This article summarizes the current understanding on the biology and the mechanisms underlying different clinical features of gastric cancers. Particularly, after a comprehensive literature search, we speculate whether specific molecular subsets of patients could derive more benefit from immunotherapy. Expert commentary: Most cancers may evade the immune response, which is normally regulated by a delicate balance between activating and inhibitory signals. For example, both CTLA-4 and PD-1, once linked to PD-L1/2, may inhibit T-cell signaling. The use of agent to harness the power of the immune system appears to be the ultimate frontier in gastric cancer treatment. While anti-CTLA-4 antibodies are minimally active, there is growing evidence for the efficacy of PD1/-L1 inhibitors. The search of predictive factors for immunotherapy will provide key hints towards the optimal use of these agents.


NMR spectroscopic study of the self-aggregation of 3-hexen-1,5-diyne derivatives.
The self-assembly of polycatenar molecules derived from 1,6-diphenyl-3,4-dipropyl-3-hexen-1,5-diyne has been studied in detail by solution NMR spectroscopy. The analysis of the concentration- and temperature-dependent evolution of the chemical shifts and the diffusion coefficients in [D12]cyclohexane agrees well with an isodesmic model of association in this solvent. The association constants for the stacking and entropy and enthalpy of the process have been obtained. The driving force for the aggregation process is provided by a negative enthalpy (ΔH), which is partially compensated by a negative entropy (ΔS). A structural study of the self-assembly in solution has been carried out with the help of NOESY NMR spectroscopic experiments.


Comparison of clinical outcomes between androgen deprivation therapy with up-front abiraterone and bicalutamide for Japanese patients with LATITUDE high-risk prostate cancer in a real-world retrospective analysis.
Combining abiraterone (Abi) with androgen deprivation therapy (ADT) improves overall survival, compared to ADT only, in patients with metastatic castration-sensitive prostate cancer (mCSPC). In Japan, bicalutamide (Bica) and ADT (combined androgen blockade: CAB) is frequently provided for mCSPC. Because these two treatments have not been compared, mCSPC patients who received either treatment were retrospectively analyzed. Of 178 patients with LATITUDE high-risk mCSPC, 103 had received ADT plus upfront Abi (Abi group) and 75 had received ADT plus Bica (Bica group) in multiple institutions of the Tokai Urologic Oncology Research Seminar. Kaplan-Meir curves were used to retrospectively analyze survival and cancer recurrence. Univariate and multivariate Cox regression analyses identified potential prognostic factors for progression-free survival (PFS). Significant differences in major clinicopathological characteristics between the two groups were not observed. The rate of castration-resistant development was higher in the Bica compared to Abi group (50.6 vs. 25.2%, p < 0.001). The median PFS in the Bica group was 13.6 months {95% confidence interval [CI] 9.2-22.2}; however, in the Abi group, PFS did not reach the median {95% CI 18.5-not assessed [NA]; p < 0.001}. Time to second progression for the Abi group was superior (p = 0.07). Univariate and multivariate analyses revealed Gleason pattern 5, high alkaline phosphatase levels, and conventional CAB using Bica as significant prognostic factors for short PFS. In patients with LATITUDE high-risk mCSPC, upfront use of Abi combined with ADT resulted in favorable prognostic outcomes compared with conventional ADT with Bica.


Toxicity of essential and non-essential oils against the chewing louse, Bovicola (Werneckiella) ocellatus.
The toxicity of six plant essential oils to the chewing louse, Bovicola (Werneckiella) ocellatus collected from donkeys, was examined in laboratory bioassays. The oils examined were: tea-tree (Melaleuca alternifolia), lavender (Lavandula angustifolia), peppermint (Mentha piperita), eucalyptus (Eucalyptus globulus Labillardiere), clove bud (Eugenia caryophyllata) and camphor (Cinnamomum camphora). All except camphor oil showed high levels of toxicity, with significant dose-dependent mortality and an LC(50) at concentrations of below 2% (v/v). Hundred percent mortality was achieved at concentrations of 5-10% (v/v). Two essential oil components: eugenol and (+)-terpinen-4-ol showed similar levels of toxicity. The data suggest that these botanical products may offer environmentally and toxicologically safe, alternative veterinary pediculicides for the control of ectoparasitic lice.


Utilization Patterns of a Food Referral Program: Findings from the Mid-Ohio Farmacy.
There is limited evidence describing utilization of clinic-based food referral programs intended to support healthy eating for food-insecure patients. To address this gap, this study aims to describe the utilization of the Mid-Ohio Farmacy (MOF). MOF is a partnership between a regional foodbank and local health care providers, including an academic medical center (AMC), that enables referrals of patients that experience food insecurity to a network of participating food pantries. This observational study uses data from 2 AMC family medicine clinics that offered the MOF referral from September 2019 to November 2020. Patients who screened positive for food insecurity and had an eligible chronic disease (eg, diabetes, hypertension, obesity) were referred to the MOF. We compared demographic and clinical characteristics of patients that filled their referral (ie, visitors) to those that did not (ie, non-visitors). Among visitors, we also assessed patterns of pantry utilization. In total, 51% (164 of 322) of patients referred to the MOF visited a food pantry at least once. Visitors were more likely to be older, have diabetes, and have visited a food pantry before their referral. Patients with uncontrolled hypertension were less likely to visit a food pantry following their referral. Patients that had visited a food pantry before their referral had more visits in total and more produce-specific visits following their referral. Our results suggest that while the MOF can connect patients to food resources, further attention may be needed to encourage its use among patients who have not previously accessed pantries.


Expression profiling of nucleotide metabolism-related genes in human breast cancer cells after treatment with 5-fluorouracil.
5-Fluorouracil (5-FU) is one of the most widely used drugs for treatment of cancers, including breast cancer that exhibits its anticancer activity by inhibiting DNA synthesis and also incorporated into DNA and RNA. The objective of this investigation was to find out the total nucleotide metabolism genes regulated by 5-FU in breast cancer cell line. The breast cancer cell line MCF-7 was treated with the drug 5-FU. To analyze the expression of genes, we have conducted the experiment using 1.7 k and 19k human microarray slide and confirmed the expression of genes by semiquantitative reverse transcription-polymerase chain reaction. The expression of 44 genes involved in the nucleotide metabolism pathway was quantified. Of these 44 genes analyzed, transcription of 6 genes were upregulated and 9 genes were downregulated. Earlier studies revealed that the transcription of genes for key enzymes like thymidylate synthase, thymidine kinase, and dihydropyrimidine dehydrogenase are regulated by 5-FU. This study identified some novel genes like thioredoxin reductase, ectonucleotide triphosphate dephosphorylase, and CTP synthase are regulated by 5-FU. The data also reveal large-scale perturbation in transcription of genes not involved directly in the known mechanism of action of 5-FU.


Human evolution, life history theory, and the end of biological reproduction.
Throughout primate history there have been three major life history transitions towards increasingly delayed sexual maturation and biological reproduction, as well as towards extended life expectancy. Monkeys reproduce later and live longer than do prosimians, apes reproduce later and live longer than do monkeys, and humans reproduce later and live longer than do apes. These life history transitions are connected to increased encephalization. During the last life history transition from apes to humans, increased encephalization co-evolved with increased dependence on cultural knowledge for energy acquisition. This led to a dramatic pressure for more energy investment in growth over current biological reproduction. Since the industrial revolution socioeconomic development has led to even more energy being devoted to growth over current biological reproduction. I propose that this is the beginning of an ongoing fourth major primate life history transition towards completely delayed biological reproduction and an extension of the evolved human life expectancy. I argue that the only fundamental difference between this primate life history transition and previous life history transitions is that this transition is being driven solely by cultural evolution, which may suggest some deeper evolutionary transition away from biological evolution is already in the process of occurring.


Cardiovascular sex differences influencing microvascular exchange.
The vital role of the cardiovascular (CV) system is maintenance of body functions via the matching of exchange to tissue metabolic demand. Sex-specific differences in the regulatory mechanisms of CV function and the metabolic requirements of men and women, respectively, have been identified and appreciated. This review focuses on sex differences of parameters influencing exchange at the point of union between blood and tissue, the microvasculature. Microvascular architecture, blood pressure (hydrostatic and oncotic), and vascular permeability, therefore, are discussed in the specific context of sex in health and disorders. It is notable that when sex differences exist, they are generally subtle but significant. In the aggregate, though, they can give rise to profoundly different phenotypes. The postulated mechanisms responsible for sex differences are attributed to genomics, epigenetics, and sex hormones. Depending on specific circumstances, the effect of the combined factors can range from insignificant to lethal. Identifying and understanding key signalling mechanisms bridging genomics/sex hormones and microvascular exchange properties within the scope of this review holds significant promise for sex-specific prevention and treatment of vascular barrier dysfunction.


Polyphenols from Root, Tubercles and Grains Cropped in Brazil: Chemical and Nutritional Characterization and Their Effects on Human Health and Diseases.
Throughout evolution, plants have developed the ability to produce secondary phenolic metabolites, which are important for their interactions with the environment, reproductive strategies and defense mechanisms. These (poly)phenolic compounds are a heterogeneous group of natural antioxidants found in vegetables, cereals and leguminous that exert beneficial and protective actions on human health, playing roles such as enzymatic reaction inhibitors and cofactors, toxic chemicals scavengers and biochemical reaction substrates, increasing the absorption of essential nutrients and selectively inhibiting deleterious intestinal bacteria. Polyphenols present in some commodity grains, such as soy and cocoa beans, as well as in other vegetables considered security foods for developing countries, including cassava, taro and beetroot, all of them cropped in Brazil, have been identified and quantified in order to point out their bioavailability and the adequate dietary intake to promote health. The effects of the flavonoid and non-flavonoid compounds present in these vegetables, their metabolism and their effects on preventing chronic and degenerative disorders like cancers, diabetes, osteoporosis, cardiovascular and neurological diseases are herein discussed based on recent epidemiological studies.


Granzyme B is not required for regulatory T cell-mediated suppression of graft-versus-host disease.
Regulatory T (T(reg)) cells can suppress a wide variety of immune responses, including antitumor and alloimmune responses. The mechanisms by which T(reg) cells mediate their suppressive effects depend on the context of their activation. We previously reported that granzyme B is important for T(reg) cell-mediated suppression of antitumor immune responses. We therefore hypothesized that granzyme B may likewise be important for suppression of graft-versus-host disease (GVHD). We found that allogeneic mismatch induces the expression of granzyme B in mixed lymphocyte reactions and in a model of graft-versus-host disease (GVHD). However, wild-type and granzyme B-deficient T(reg) cells were equally able to suppress effector T (T(eff)) cell proliferation driven by multiple stimuli, including allogeneicantigen-presenting cells. Surprisingly, adoptive transfer of granzyme B-deficient T(reg) cells prevented GVHD lethality, suppressed serum cytokine production in vivo, and prevented target organ damage. These data contrast strikingly with our previous study, which demonstrated that granzyme B plays a nonredundant role in T(reg) cell-mediated suppression of antitumor responses. Taken together, these findings suggest that targeting specific T(reg) cell-suppressive mechanisms, such as granzyme B, may be therapeutically beneficial for segregating GVHD and graft-versus-tumor immune responses.


The RNA methyltransferase METTL8 installs m3C32 in mitochondrial tRNAsThr/Ser(UCN) to optimise tRNA structure and mitochondrial translation.
Modified nucleotides in tRNAs are important determinants of folding, structure and function. Here we identify METTL8 as a mitochondrial matrix protein and active RNA methyltransferase responsible for installing m3C32 in the human mitochondrial (mt-)tRNAThr and mt-tRNASer(UCN). METTL8 crosslinks to the anticodon stem loop (ASL) of many mt-tRNAs in cells, raising the question of how methylation target specificity is achieved. Dissection of mt-tRNA recognition elements revealed U34G35 and t6A37/(ms2)i6A37, present concomitantly only in the ASLs of the two substrate mt-tRNAs, as key determinants for METTL8-mediated methylation of C32. Several lines of evidence demonstrate the influence of U34, G35, and the m3C32 and t6A37/(ms2)i6A37 modifications in mt-tRNAThr/Ser(UCN) on the structure of these mt-tRNAs. Although mt-tRNAThr/Ser(UCN) lacking METTL8-mediated m3C32 are efficiently aminoacylated and associate with mitochondrial ribosomes, mitochondrial translation is mildly impaired by lack of METTL8. Together these results define the cellular targets of METTL8 and shed new light on the role of m3C32 within mt-tRNAs.


The assessment of segmental healing by the Modified Mayo Endoscopic Score (MMES) complements the prediction of long-term clinical outcomes in patients with ulcerative colitis.
Current endoscopic scoring systems for ulcerative colitis (UC) do not consider the extent of mucosal inflammation. The modified Mayo endoscopic score (MMES) was developed to detect segmental endoscopic improvement. We evaluated the ability of the MMES to predict long-term clinical outcomes and compared it to the widely used Mayo endoscopic subscore (MES). Consecutive patients with moderate to severe UC starting biological therapy were enrolled between January 2014 and September 2017 in this prospective observational study. A clinical and endoscopic evaluation was performed at baseline and at week 8/14. A modified Mayo score was used to grade clinical activity, MES and MMES were used to evaluate endoscopic activity. Patients were divided into 3 groups according to the evolution of endoscopic activity, namely endoscopic improvement (MES ≤ 1), segmental endoscopic response only (MES > 1, but decrease in MMES ≥ 30%) or no endoscopic response (all others). Over the follow-up period clinical relapse-, discontinuation- and colectomy-free survival were assessed. A total of 150 patients were included (48% female, median age 42 years, median disease duration 7 years) with a median follow-up of 61 months. We identified 69 patients with endoscopic improvement, 27 with segmental endoscopic response and 54 without endoscopic response. Patients with segmental endoscopic response showed intermediate long-term clinical outcomes as compared to the other two groups (log rank p = 0.003 for clinical relapse-, and p < 0.001 for both discontinuation- and colectomy-free survival). The MMES exhibited a benefit in predicting long-term outcome in UC even though endoscopic improvement remains the strongest predictor.


Incidence, risk factors and prevention of mild traumatic brain injury: results of the WHO Collaborating Centre Task Force on Mild Traumatic Brain Injury.
We undertook a best-evidence synthesis on the incidence, risk factors and prevention of mild traumatic brain injury. Medline, Cinahl, PsycINFO and Embase were searched for relevant articles. After screening 38,806 abstracts, we critically reviewed 169 studies on incidence, risk and prevention, and accepted 121 (72%). The accepted articles show that 70-90% of all treated brain injuries are mild, and the incidence of hospital-treated patients with mild traumatic brain injury is about 100-300/100,000 population. However, much mild traumatic brain injury is not treated at hospitals, and the true population-based rate is probably above 600/100,000. Mild traumatic brain injury is more common in males and in teenagers and young adults. Falls and motor-vehicle collisions are common causes. Strong evidence supports helmet use to prevent mild traumatic brain injury in motorcyclists and bicyclists. The mild traumatic brain injury literature is of varying quality, and the studies are very heterogeneous. Nevertheless, there is evidence that mild traumatic brain injury is an important public health problem, but we need more high-quality research into this area.


Studies on the bacteriophage PS8 of Agrobacterium tumefaciens (Smith and Townsend) Conn: physico-chemical properties of its DNA.
DNA from the bacteriophage PS8 was extracted and purified. The buoyant density was determined was 1.716 cm3/g. The guanine-cytosine content was calculated to be 57%. DNA molecules which looked like circles were found among linear strands in an electron-microscopic study. With an endonuclease from Streptomyces albus G the DNA was digested to 19 fragments, with molecular weights ranging from 600 to 7,400 daltons. The molecular weight of the DNA was determined to be 38.8 X 10(6) daltons +/- 8.7%.


Air pollution and daily mortality in Rome, Italy.
To assess the relation between several daily indicators of air pollution (particulates and gases) and daily mortality in the metropolitan area of Rome and in the central part of the city. Time series analysis. The associations between daily concentrations of pollutants (particles, SO2, NO2, CO, O3) recorded by five fixed monitors and daily total mortality in the period from January 1992 to June 1995 were evaluated. The analysis included examination of the pollution effect on mortality by place of residence within the metropolitan area, by season, age, place of death (in and out a hospital), and cause of death (cardiovascular and respiratory disease). The Poisson model included loses smooth functions of the day of study, mean temperature, mean humidity, and indicator variables for day of the week and holidays. The mean daily number of deaths was 56.9 (44.8 among people > or = 65 years old). A mean of 36.3 deaths occurred in the city centre; 37.3 deaths a day were recorded in a hospital. Total mortality was significantly associated with a 10 micrograms/m3 increase in particles (0.4%) on that day (log 0), and with a 10 micrograms/m3 increase in NO2 at lag 1 (0.3%) and lag 2 (0.4%) (1 and 2 days before, respectively). The effect of particles (lag 0) and of NO2 (lag 2) on total mortality was higher among those living in the city centre (0.7% and 0.5%, respectively). The risk estimates were higher in the warmer season (1.0% and 1.1%, respectively), whereas no difference was found for those dying in or out of the hospital. The effect of particles was robust to a sensitivity analysis and to the inclusion of NO2 in the regression model. Increase in particulates and NO2, generated by the same mobile combustion sources, is associated with a short term increase in mortality in Rome. The effect is more evident among residents in the city centre, where the levels of exposure to pollutants recorded by fixed monitors are probably more reliable indicators of personal exposure.


A Vaccination Simulator for COVID-19: Effective and Sterilizing Immunization Cases.
In this work, we present a particle-based SEIR epidemic simulator as a tool to assess the impact of different vaccination strategies on viral propagation and to model sterilizing and effective immunization outcomes. The simulator includes modules to support contact tracing of the interactions amongst individuals and epidemiological testing of the general population. The particles are distinguished by age to represent more accurately the infection and mortality rates. The tool can be calibrated by region of interest and for different vaccination strategies to enable locality-sensitive virus mitigation policy measures and resource allocation. Moreover, the vaccination policy can be simulated based on the prioritization of certain age groups or randomly vaccinating individuals across all age groups. The results based on the experience of the province of Lecco, Italy, indicate that the simulator can evaluate vaccination strategies in a way that incorporates local circumstances of viral propagation and demographic susceptibilities. Further, the simulator accounts for modeling the distinction between sterilizing immunization, where immunized people are no longer contagious, and effective immunization, where the individuals can transmit the virus even after getting immunized. The parametric simulation results showed that the sterilizing-age-based vaccination scenario results in the least number of deaths. Furthermore, it revealed that older people should be vaccinated first to decrease the overall mortality rate. Also, the results showed that as the vaccination rate increases, the mortality rate between the scenarios shrinks.


Synthesis, characterization, thermal behavior, and DNA-cleaving studies of cyano-bridged nickel(II)-copper(II) complexes of 4-(pyridin-2-ylazenyl)resorcinol.
We present here the syntheses of a mononuclear Cu(II) complex and two polynuclear Cu(II)-Ni(II) complexes of the azenyl ligand, 4-(pyridin-2-ylazenyl)resorcinol (HL; 1). The reaction of HL (1) and copper(II) perchlorate with KCN gave a mononuclear complex [CuL(CN)] (4). Using 4, one pentanuclear complex, [{CuL(NC)}(4) Ni](ClO(4))(2) (5) and one trinuclear complex, [{CuL(CN)}(2) NiL]ClO(4) (6), were prepared and characterized by elemental analyses, magnetic susceptibility, molar conductance, IR, and thermal analysis. Stoichiometric and spectral results of the mononuclear Cu(II) complex indicated that the metal/ligand/CN ratio was 1 : 1 : 1, and the ligand behaved as a tridentate ligand forming neutral metal chelates through the pyridinyl and azenyl N-, and resorcinol O-atom. The interaction between the compounds (the ligand 1, its Ni(II) and Cu(II) complexes without CN, i.e., 2 and 3, and its complexes with CN, 4-6) and DNA has also been investigated by agarose gel electrophoresis. The pentanuclear Cu(4) Ni complex (5) with H(2) O(2) as a co-oxidant exhibited the strongest DNA-cleaving activity.


Activation of the Notch pathway in the hair cortex leads to aberrant differentiation of the adjacent hair-shaft layers.
Little is known about the mechanisms underlying the generation of various cell types in the hair follicle. To investigate the role of the Notch pathway in this process, transgenic mice were generated in which an active form of Notch1 (Notch(DeltaE)) was overexpressed under the control of the mouse hair keratin A1 (MHKA1) promoter. MHKA-Notch(DeltaE) is expressed only in one precursor cell type of the hair follicle, the cortex. Transgenic mice could be easily identified by the phenotypes of curly whiskers and wavy, sheen pelage hair. No effects of activated Notch on proliferation were detected in hair follicles of the transgenic mice. We find that activating Notch signaling in the cortex caused abnormal differentiation of the medulla and the cuticle, two neighboring cell types that did not express activated Notch. We demonstrate that these non-autonomous effects are likely caused by cell-cell interactions between keratinocytes within the hair follicle and that Notch may function in such interactions either by directing the differentiation of follicular cells or assisting cells in interpreting a gradient emanating from the dermal papilla.


SNORA21 - An Oncogenic Small Nucleolar RNA, with a Prognostic Biomarker Potential in Human Colorectal Cancer.
Emerging evidence indicates that small nucleolar RNAs (snoRNAs) play a central role in oncogenesis. Herein, we systematically evaluated expression profiles of snoRNAs in colorectal cancer (CRC) and investigated their clinical and functional role in this malignancy. We compared expression levels of snoRNAs between cancer and normal tissues using publicly available datasets and identified the most differentially expressed and commonly upregulated snoRNAs in CRC. These results were examined in 489 colorectal tissues to assess their clinical significance, followed by a series of in vitro and in vivo experiments to evaluate the functional role of candidate snoRNAs. Using multiple RNA profiling datasets, we identified consistent overexpression of SNORA21 in CRC. In the clinical validation cohorts, the expression level of SNORA21 was upregulated in colorectal adenomas and cancers. Furthermore, elevated SNORA21 emerged as an independent factor for predicting poor survival. Both in vitro and in vivo experiments revealed that CRISPR/Cas9-mediated inhibition of SNORA21 expression resulted in decreased cell proliferation and invasion through modulation of multiple cancer related pathways. We systematically identified SNORA21 as a key oncogenic snoRNA in CRC, which plays an important role in cancer progression, and might serve as an important prognostic biomarker in CRC.


Resources used by general practitioners for advising travelers from New Zealand.
The risks of the destination and any specific requirements for travel health advice may be obtained from a variety of resources. This study was designed to investigate the usefulness of various resources available in New Zealand for providing travel health advice and the extent to which GPs used these resources in providing travel health advice. Four hundred GPs (400/2830) were randomly selected from the register of the New Zealand Medical Council and sent self-administered questionnaires. Two reminders were sent. Three hundred and thirty-two (332/400, 83%) GPs responded. The usefulness of various resources was reported, including Health Advice for Overseas Travellers (277/289, 96%), New Ethicals (256/278, 92%), New Zealand Public Health Report (79/164, 48%), International Travel and Health (41/144, 28%), computerized databases (6/122, 5%), journals (14/130, 11%), and other resources (44/139, 32%). Health Advice for Overseas Travellers was regarded as significantly more useful than International Travel and Health (x2 = 4,68, df = 1, p <.05). Only 23% (70/309) of respondents indicated that they always used these resources in their practice of travel medicine. Fifty percent (154/309) of respondents indicated that they usually used these resources, while 27% (83/309) of respondents indicated that they used these resources sometimes. Only 1% (2/309) of GPs did not use resources at all for their practice of travel medicine. The most useful resource was Health Advice for Overseas Travellers, which outlines the New Zealand recommendations for medical practitioners providing travel health advice. It may be useful for GPs to gain access to and training in association with a greater range of specialist resources to use in conjunction with the provision of travel health advice. These might include international guidelines, journals, and access to computerized databases and the internet. With the recent introduction of a widely accessible computerized database in New Zealand, follow-up studies could be instituted to determine if GPs' use of computerized databases becomes more widespread and whether access to and use of these computerized databases influences the provision of travel health advice by GPs. Further studies are needed to examine the appropriateness of the advice provided by the various resources used by GPs in New Zealand.


Combining attention training with cognitive-behavior therapy in Internet-based self-help for social anxiety: study protocol for a randomized controlled trial.
Guided Internet-based cognitive-behavioral therapy (ICBT) has been found to be effective for social anxiety disorder (SAD) by several independent research groups. However, since the extent of clinically significant change demonstrated leaves room for improvement, new treatments should be developed and investigated. A novel treatment, which has generally been found to be effective, is cognitive bias modification (CBM). This study aims to evaluate the combination of CBM and ICBT. It is intended that two groups will be compared; one group randomized to receiving ICBT and CBM towards threat cues and one group receiving ICBT and control training. We hypothesize that the group receiving ICBT plus CBM will show superior treatment outcomes. Participants with SAD (N = 128), will be recruited from the general population. A composite score combining the scores obtained from three social anxiety questionnaires will serve as the primary outcome measure. Secondary measures include self-reported depression and quality of life. All treatments and assessments will be conducted via the Internet and measurement points will be baseline, Week 2, post-treatment, and 4 months post-treatment. There is no direct evidence of the effects of combining CBM and ICBT in SAD. Adding attention-training sessions to ICBT protocols could increase the proportion of participants who improve and recover through Internet-based self-help. ClinicalTrials.gov: NCT01570400.


Identification of cryptic products of the gliotoxin gene cluster using NMR-based comparative metabolomics and a model for gliotoxin biosynthesis.
Gliotoxin, a major product of the gli non-ribosomal peptide synthetase gene cluster, is strongly associated with virulence of the opportunistic human pathogen Aspergillus fumigatus. Despite identification of the gli cluster, the pathway of gliotoxin biosynthesis has remained elusive, in part because few potential intermediates have been identified. In addition, previous studies suggest that knowledge of gli-dependent metabolites is incomplete. Here we use differential analysis by 2D NMR spectroscopy (DANS) of metabolite extracts derived from gli knock-out and wild-type (WT) strains to obtain a detailed inventory of gli-dependent metabolites. DANS-based comparison of the WT metabolome with that of ΔgliZ, a knock-out strain devoid of the gene encoding the transcriptional regulator of the gli cluster, revealed nine novel gliZ-dependent metabolites including unexpected structural motifs. Their identification provides insight into gliotoxin biosynthesis and may benefit studies of the role of the gli cluster in A. fumigatus virulence. Our study demonstrates the utility of DANS for correlating gene expression and metabolite biosynthesis in microorganisms.


Thymectomy for myasthenia gravis.
The results of 14 years' experience in the surgical treatment of myasthenia gravis are reported. Twenty-one patients (14 female, 7 male) underwent thymectomy for myasthenia gravis between 1971 and 1984. The mean age of the patients was 33 years (range 14 - 57 years). The median duration of symptoms prior to surgery was 18 months (range 5 months to 35 years). The mean follow-up was 5.3 years. There were no post-operative deaths: 76% obtained benefit from thymectomy. The patients' age, sex, duration of symptoms and histology of the thymus gland did not correlate with the result of treatment. This series suggests that, while thymectomy is often beneficial in the treatment of myasthenia gravis, there are no accurate predictors of the outcome following surgery.


Pathogenicity and virulence of Shigella sonnei: A highly drug-resistant pathogen of increasing prevalence.
Shigella spp. are the causative agent of shigellosis (or bacillary dysentery), a diarrhoeal disease characterized for the bacterial invasion of gut epithelial cells. Among the 4 species included in the genus, Shigella flexneri is principally responsible for the disease in the developing world while Shigella sonnei is the main causative agent in high-income countries. Remarkably, as more countries improve their socioeconomic conditions, we observe an increase in the relative prevalence of S. sonnei. To date, the reasons behind this change in aetiology depending on economic growth are not understood. S. flexneri has been widely used as a model to study the pathogenesis of the genus, but as more research data are collected, important discrepancies with S. sonnei have come to light. In comparison to S. flexneri, S. sonnei can be differentiated in numerous aspects; it presents a characteristic O-antigen identical to that of one serogroup of the environmental bacterium Plesiomonas shigelloides, a group 4 capsule, antibacterial mechanisms to outcompete and displace gut commensal bacteria, and a poorer adaptation to an intracellular lifestyle. In addition, the World Health Organization (WHO) have recognized the significant threat posed by antibiotic-resistant strains of S. sonnei, demanding new approaches. This review gathers knowledge on what is known about S. sonnei within the context of other Shigella spp. and aims to open the door for future research on understanding the increasing spread of this pathogen.


TSH levels and risk of miscarriage in women on long-term levothyroxine: a community-based study.
Thyroid dysfunction is associated with adverse obstetric outcomes, but there is limited information on pregnancy outcomes in women established on levothyroxine. The objective of the study was to determine the relationship between TSH levels and pregnancy outcomes in levothyroxine-treated women in a large community-based database. This was a historical cohort analysis. Individuals with a first prescription of levothyroxine from 2001 through 2009 (n = 55 501) were identified from the UK General Practice Research Database (population 5 million). Of these, we identified 7978 women of child-bearing age (18-45 y) and 1013 pregnancies in which levothyroxine had been initiated at least 6 months before conception. TSH, miscarriage/delivery status, and obstetric outcomes were measured. Forty-six percent of levothyroxine-treated women aged 18-45 years had a TSH level greater than 2.5 mU/L (recommended upper level in the first trimester). Among pregnant women who had their TSH measured in the first trimester, 62.8% had a TSH level greater than 2.5 mU/L, with 7.4% greater than 10 mU/L. Women with TSH greater than 2.5 mU/L in the first trimester had an increased risk of miscarriage compared with women with TSH 0.2-2.5 mU/L after adjusting for age, year of pregnancy, diabetes, and social class (P = .008). The risk of miscarriage was increased in women with TSH 4.51-10 mU/L [odds ratio (OR) 1.80, 95% confidence interval (CI) 1.03, 3.14)] and TSH greater than 10 mU/L (OR 3.95, 95% CI 1.87, 8.37) but not with TSH 2.51-4.5 mU/L (OR 1.09, 95% CI 0.61, 1.93). The majority of levothyroxine-treated women have early gestational TSH levels above the recommended targets (>2.5 mU/L) with a strong risk of miscarriage at levels exceeding 4.5 mU/L. There is an urgent need to improve the adequacy of thyroid hormone replacement in early pregnancy.


[Prolactin. New clinical importance of a phylogenetically old hormone].
New clinical significance of a phylogenetically old hormone. Recently, human Prolactin(hPRL) has been established as an anterior pituitary hormone which can be separated from growth hormone. By radioimmunoassay of hPRL it could be shown that many pituitary tumors formerly thought to be hormonally inactive, were in fact producing prolactin. Hyperprolactinaemia, with and without pituitary tumors, causing amenorrhea and galactorrhea in females, and loss of libido and impotence in males, can be treated effectively by neurosurgical procedures in tumor cases, or by medical inhibition of hPRL-secretion with bromocryptin.


Effects of voluntary contraction on descending volleys evoked by transcranial stimulation in conscious humans.
1. The spinal volleys evoked by single transcranial magnetic or electric stimulation over the cerebral motor cortex were recorded from a bipolar electrode inserted into the cervical epidural space of three conscious human subjects. These volleys were termed direct (D) and indirect (I) waves according to their latency. 2. We measured the size and number of volleys elicited by magnetic stimulation at various intensities with subjects at rest and during 20 or 100 % maximum contraction of the contralateral first dorsal interosseous muscle (FDI). Surface EMG activity was also recorded. 3. Electrical stimulation evoked a D-wave volley. Magnetic stimulation at intensities up to about 15 % of stimulator output above threshold evoked only I-waves. At higher intensities, a D-wave could be seen in two of the three subjects. 4. At all intensities tested, voluntary contraction increased the number and size of the I-waves, particularly during maximum contractions. However, there was only a small effect on the threshold for evoking descending activity. Voluntary contraction produced large changes in the size of EMG responses recorded from FDI. 5. Because the recorded epidural activity is destined for muscles other than the FDI, it is impossible to say to what extent increased activity contributes to voluntary facilitation of EMG responses. Indeed, our results suggest that the main factor responsible for enhancing EMG responses in the transition from rest to activity is likely to be increased excitability of spinal motoneurones, rather than increases in the corticospinal volley. The latter may be more important in producing EMG facilitation at different levels of voluntary contraction.


Localization of two interchain disulfide bridges in dimers of bovine alpha s2-casein. Parallel and antiparallel alignments of the polypeptide chains.
Carboxymethylation of bovine skimmed milk with 14C-labelled iodoacetic acid followed by purification of the alpha s2-casein dimer showed that all four cysteine residues in the protein are engaged in disulfide linkages. Mass spectrometry and sequence analysis of cystine-containing tryptic peptides revealed the presence of two interchain disulfide bridges in the protein. Sequence analysis of disulfide-linked peptides resulting from an enzymatic cleavage between the bridges demonstrated that the individual chains in the dimers are either aligned in an antiparallel or a parallel orientation. The identity of some of the disulfide-linked peptides was further verified by performic acid oxidation followed by sequence analysis of the resulting peptides.


The neural network of the basal ganglia as revealed by the study of synaptic connections of identified neurones.
The study of synaptic connections in the electron microscope has established an 'elementary' circuit for the neostriatum which consists of a pathway from cortical areas (neocortex, hippocampus, amygdala) to medium spiny neurones of the striatum that also receive converging synaptic input from midbrain dopamine neurones. The striatal medium spiny neurones are projection neurones and they form synaptic contacts with output neurones in the globus pallidus and substantia nigra reticulata. In this way, dopaminergic afferents can directly modulate the flow of information from cortical areas through the striatum to the 'premotor' areas of the brainstem and to the thalamus. It is proposed that certain parts of the striatum can themselves control the activity of midbrain dopamine neurones and so one part of the striatum can 'gate' the flow of information through another part.


The rise, the fall and the renaissance of vitamin E.
This review deals with the expectations of vitamin E ability of preventing or curing, as a potent antioxidant, alleged oxidative stress based ailments including cardiovascular disease, cancer, neurodegenerative diseases, cataracts, macular degeneration and more. The results obtained with clinical intervention studies have highly restricted the range of effectiveness of this vitamin. At the same time, new non-antioxidant mechanisms have been proposed. The new functions of vitamin E have been shown to affect cell signal transduction and gene expression, both in vitro and in vivo. Phosphorylation of vitamin E, which takes place in vivo, results in a molecule provided with functions that are in part stronger and in part different from those of the non-phosphorylate compound. The in vivo documented functions of vitamin E preventing the vitamin E deficiency ataxia (AVED), slowing down the progression of non-alcoholic steato-hepatitis (NASH), decreasing inflammation and potentiating the immune response are apparently based on these new molecular mechanisms. It should be stressed however that vitamin E, when present at higher concentrations in the body, should exert antioxidant properties to the extent that its chromanol ring is unprotected or un-esterified.


The mean and variance of the selection differential after top-down selection from a mixture of normal samples.
A procedure is presented for determining the mean and variance of the selection differential for top-down selections in which the candidates come from populations that have a different average score on the selection measure. Although the procedure is based on the same stochastic model and requires identical data to the currently available method for estimating the mean selection differential, it has the advantage that the resulting expressions are valid for finite-sample selection decisions and that the variance of the selection differential can also be assessed. The difference between the two procedures is illustrated by means of an example application, and it is shown how the present results are particularly helpful in determining the expected utility of personnel selection decisions.


Entrectinib: A New Selective Tyrosine Kinase Inhibitor Approved for the Treatment of Pediatric and Adult Patients with NTRK Fusionpositive, Recurrent or Advanced Solid Tumors.
Entrectinib is a highly potent ATP-competitive and selective inhibitor of tyrosine kinases - Trk A B C, ALK, and ROS1. It was developed by Roche and initially approved in Japan in 2019 to treat pediatric and adult patients with NTRK fusionpositive, recurrent, or advanced solid tumors. In August 2019, entrectinib received accelerated approval by the U.S FDA for this indication. It is also the first FDA-approved drug designed to target both NTRK and ROS1. We aim to summarize recent studies related to the synthesis, mechanism of action, and clinical trials of the newly approved selective tyrosine kinase inhibitor entrectinib. We conduct a literature review of the research studies on the new highly-potent small-molecule entrectinib. Entrectinib, based on three clinical studies (ALKA, STARTRK-1, and STARTRK-2), was well tolerated, with a manageable safety profile. It induced clinically meaningful responses in recurrent or advanced solid tumors associated with NTRK fusion- positive or ROS1+ NSCLC. It demonstrated substantial efficacy in patients with CNS metastases.


Genetic Influences on Metabolite Levels: A Comparison across Metabolomic Platforms.
Metabolomic profiling is a powerful approach to characterize human metabolism and help understand common disease risk. Although multiple high-throughput technologies have been developed to assay the human metabolome, no technique is capable of capturing the entire human metabolism. Large-scale metabolomics data are being generated in multiple cohorts, but the datasets are typically profiled using different metabolomics platforms. Here, we compared analyses across two of the most frequently used metabolomic platforms, Biocrates and Metabolon, with the aim of assessing how complimentary metabolite profiles are across platforms. We profiled serum samples from 1,001 twins using both targeted (Biocrates, n = 160 metabolites) and non-targeted (Metabolon, n = 488 metabolites) mass spectrometry platforms. We compared metabolite distributions and performed genome-wide association analyses to identify shared genetic influences on metabolites across platforms. Comparison of 43 metabolites named for the same compound on both platforms indicated strong positive correlations, with few exceptions. Genome-wide association scans with high-throughput metabolic profiles were performed for each dataset and identified genetic variants at 7 loci associated with 16 unique metabolites on both platforms. The 16 metabolites showed consistent genetic associations and appear to be robustly measured across platforms. These included both metabolites named for the same compound across platforms as well as unique metabolites, of which 2 (nonanoylcarnitine (C9) [Biocrates]/Unknown metabolite X-13431 [Metabolon] and PC aa C28:1 [Biocrates]/1-stearoylglycerol [Metabolon]) are likely to represent the same or related biochemical entities. The results demonstrate the complementary nature of both platforms, and can be informative for future studies of comparative and integrative metabolomics analyses in samples profiled on different platforms.


Automatic adaptive system dialysis for hemodialysis-associated hypotension and intolerance: a noncontrolled multicenter trial.
Hemodialysis is complicated by a high incidence of intradialytic hypotension and disequilibrium symptoms caused by hypovolemia and a decrease in extracellular osmolarity. Automatic adaptive system dialysis (AASD) is a proprietary dialysis system that provides automated elaboration of dialysate and ultrafiltration profiles based on the prescribed decrease in body weight and sodium content. A noncontrolled (single arm), multicenter, prospective, clinical trial. 55 patients with intradialytic hypotension or disequilibrium syndrome in 15 dialysis units were studied over a 1-month interval using standard treatment (642 sessions) followed by 6 months using AASD (2,376 sessions). AASD (bicarbonate dialysis with dialysate sodium concentration and ultrafiltration rate profiles determined by the automated procedure). Primary and major secondary outcomes were the frequency of intradialytic hypotension and symptoms (hypotensive events, headache, nausea, vomiting, and cramps), respectively. More stable intradialytic systolic and diastolic blood pressures with lower heart rate were found using AASD compared with standard treatment. Sessions complicated by hypotension decreased from 58.7% ± 7.3% to 0.9% ± 0.6% (P < 0.001). The incidence of other disequilibrium syndrome symptoms was lower in patients receiving AASD. There were no differences in end-session body weight, interdialytic weight gain, or presession natremia between the standard and AASD treatment periods. A noncontrolled (single arm) study, no crossover from AASD to standard treatment. This study shows the long-term clinical efficacy of AASD for intradialytic hypotension and disequilibrium symptoms in a large number of patients and dialysis sessions.


Negative surface charge density near heart calcium channels. Relevance to block by dihydropyridines.
We have measured the density of negative surface charges near the voltage sensor for inactivation gating of (L-type) Ca channels in intact calf Purkinje fibers and in isolated myocytes from guinea pig and rat ventricles. Divalent cation-induced changes in the half-maximal voltage for inactivation were determined and were well described by curves predicted by surface potential theory. We measured shifts in inactivation induced by Ca, Sr, and Ba in the single cells, and by Sr in the Purkinje fibers. All of the data were consistent with an estimated negative surface charge density of 1 electronic charge per 250 A2. In addition, the data suggest that Ca, but neither Ba nor Sr, binds to the negative charges with an association constant on the order of 1 M-1. We find that divalent ion-induced changes in surface potential can account for most of the antagonism between these ions and Ca channel block by 1,4-dihydropyridines.


Understanding the influence of defects and surface chemistry on ferroelectric switching: a ReaxFF investigation of BaTiO3.
Ferroelectric materials such as barium titanate (BaTiO3) have a wide range of applications in nano scale electronic devices due to their outstanding properties. In this study, we developed an easily extendable atomistic ReaxFF reactive force field for BaTiO3 that can capture both its field- and temperature-induced ferroelectric hysteresis and corresponding changes due to surface chemistry and bulk defects. Using our force field, we were able to reproduce and explain a number of experimental observations: (1) the existence of a critical thickness of 4.8 nm below which ferroelectricity vanishes in BaTiO3; (2) migration and clustering of oxygen vacancies (OVs) in BaTiO3 and a reduction in the polarization and the Curie temperature due to the OVs; (3) domain wall interaction with the surface chemistry to influence the ferroelectric switching and polarization magnitude. This new computational tool opens up a wide range of possibilities for making predictions for realistic ferroelectric interfaces in energy-conversion, electronic and neuromorphic systems.


Alpha-catenin is essential in intestinal adenoma formation.
A loss-of-function mutation in the APC gene initiates colorectal carcinogenesis. Although the molecular mechanism of tumor initiation is complex, several modifier genes have been identified using mouse models, including the ApcMin mouse. Among the familial adenomatous polyposis mouse lines carrying a truncation mutation at codon 580 in Apc (Apc580D), one line (line19-Apc(580D/+)) showed a remarkably reduced incidence of intestinal adenomas (<5% compared with other lines). Extensive genetic analysis identified a deletion in the alpha-catenin (Ctnna1) gene as the cause of this suppression. Notably, the suppression only occurred when the Ctnna1 deletion was in cis-configuration with the Apc580D mutation. In all adenomas generated in line19-Apc(580D/+), somatic recombination between the Apc and Ctnna1 loci retained the wild-type Ctnna1 allele. These data strongly indicate that simultaneous inactivation of alpha-catenin and Apc during tumor initiation suppresses adenoma formation in line19-Apc(580D/+), suggesting that alpha-catenin plays an essential role in the initiation of intestinal adenomas. Although accumulating evidence obtained from human colon tumors with invasive or metastatic potential has established a tumor-suppressive role for alpha-catenin in late-stage tumorigenesis, the role of alpha-catenin in the initiation of intestinal tumorigenesis is not well documented, especially compared with that of beta-catenin. A mouse model used in this study focused on the early stage of tumor initiation and clearly indicated an essential role for alpha-catenin. Thus, alpha-catenin has dual roles in intestinal tumorigenesis, a supporting role in tumor initiation, and a suppressive role in tumor progression.


Multimodal Assessment of Mesenchymal Stem Cell Therapy for Diabetic Vascular Complications.
Peripheral arterial disease (PAD) is a debilitating complication of diabetes mellitus (DM) that leads to thousands of injuries, amputations, and deaths each year. The use of mesenchymal stem cells (MSCs) as a regenerative therapy holds the promise of regrowing injured vasculature, helping DM patients live healthier and longer lives. We report the use of muscle-derived MSCs to treat surgically-induced hindlimb ischemia in a mouse model of type 1 diabetes (DM-1). We serially evaluate several facets of the recovery process, including αVβ3 -integrin expression (a marker of angiogenesis), blood perfusion, and muscle function. We also perform microarray transcriptomics experiments to characterize the gene expression states of the MSC-treated is- chemic tissues, and compare the results with those of non-ischemic tissues, as well as ischemic tissues from a saline-treated control group. The results show a multifaceted impact of mMSCs on hindlimb ischemia. We determined that the angiogenic activity one week after mMSC treatment was enhanced by approximately 80% relative to the saline group, which resulted in relative increases in blood perfusion and muscle strength of approximately 42% and 1.7-fold, respectively. At the transcriptomics level, we found that several classes of genes were affected by mMSC treatment. The mMSCs appeared to enhance both pro-angiogenic and metabolic genes, while suppressing anti-angiogenic genes and certain genes involved in the inflammatory response. All told, mMSC treatment appears to exert far-reaching effects on the microenvironment of ischemic tissue, enabling faster and more complete recovery from vascular occlusion.


HSP60 trafficking in adult cardiac myocytes: role of the exosomal pathway.
The heat shock proteins (HSP) are a highly conserved family of proteins with critical functions in protein folding, protein trafficking, and cell signaling. These proteins also protect the cell against injury. HSP60 has been found in the extracellular space and has been identified in the plasma of some individuals. HSP60 is thought to be a "danger signal" to the immune system and is also highly immunogenic. Thus extracellular HSP60 is possibly toxic to the cell. The mechanism by which HSP60 is released into the extracellular space is unknown, as is whether it is released by cardiac myocytes. We investigated several different pathways controlling protein release including the classic, Golgi-mediated pathway. We found that HSP60 is released via exosomes, and that within the exosome, HSP60 is tightly attached to the exosome membrane.


An automated method for detection of layer activation order in information processing pathway of rat barrel cortex under mechanical whisker stimulation.
Rodents perform object localization, texture and shape discrimination very precisely through whisking. During whisking, microcircuits in corresponding barrel columns get activated to segregate and integrate tactile information through the information processing pathway. Sensory signals are projected through the brainstem and thalamus to the corresponding 'barrel columns' where different cortical layers are activated during signal projection. Therefore, having precise information about the layer activation order is desirable to better understand this signal processing pathway. This work proposes an automated, computationally efficient and easy to implement method to determine the cortical layer activation from intracortically recorded local field potentials (LFPs) and derived current source density (CSD) profiles: 1. Barrel cortex LFPs are represented by a template of four subsequent events: small positive/negative (E1) → large negative (E2) → slow positive (E3)→ slow long negative (E4). The method exploits the layer specific characteristics of LFPs to obtain latencies of the individual events (E1–E4), then taking the latency of E2 for calculating the layer activation order. 2. The corresponding CSD profile is calculated from the LFPs and the first sink's peak is considered as a reference point to calculate latencies and evaluate the layer activation order. Other reference points require manual calculation. Similar results of layer activation sequence are found using LFPs and CSDs. Extensive tests on LFPs recorded using standard borosilicate micropipettes demonstrated the method's workability. An interpretation of layer activation order and CSD profiles on the basis of a simplified interacortical barrel column architecture is also provided.