Streamlined construction of robust heteroprotein complexes by self-induced in-cell disulfide pairing.
Biomolecules and their functional subdomains are essential building blocks in the creation of multifunctional nanocomplexes. Methyl-binding domain protein 2 (MBD2) and p66α stand out as small α-helical motifs with an ability to self-assemble into a heterodimeric coiled-coil, making them promising building units. Yet, their practical use is hindered by rapid dissociation upon dilution. In this study, novel fusion tags, MBD2 and p66α variants, were developed to covalently link during co-expression in E. coli SHuffle. Through strategic placement of cysteine at each α-helix's terminus, intracellular crosslinking occurred with high specificity and yield, facilitated by preserved α-helical interactions. This instant disulfide bonding in the oxidative cytoplasm of E. coli SHuffle efficiently overcame the need for inefficient in vitro oxidation and protein extraction prone to creating non-specific adducts and suboptimal bioprocesses. In contrast to their wild-type counterparts, the GFP-mCherry protein complex cross-linked by the fusion tags maintained the heterodimeric state even under extensive dilution. The fusion tags, when combined with the E. coli SHuffle system, allowed for the streamlined co-expression of a stable protein complex through self-induced intracellular cysteine coupling. The approach demonstrated herein holds great promise for producing multifunctional and robust heteroprotein complexes.


Surge in regulatory T cells does not prevent onset of hyperglycemia in NOD mice: immune profiles do not correlate with disease severity.
Immune profiling of non-obese diabetic (NOD) is a widely employed tool to assess the mechanisms of inflammatory insulitis. Our analysis of the female NOD colony revealed similar distribution of lymphoid lineages to wild type mice, and at various ages of prediabetic and diabetic mice. The profiles of mesenteric and pancreatic lymph nodes differ and often change reciprocally due to directed migration of T cells towards the site of inflammation. Significant events in our colony include early decline in CD4(+)CD25(+)CD62L(+) Treg, accompanied by gradual increase in CD4(+)CD25(+)FoxP3(+) Treg in peripheral lymphoid organs and pancreatic infiltrates. Impressively, aged euglycemic mice display significant transient rise in CD4(+)CD25(-)FoxP3(+) Treg in the thymus, pancreas and draining lymph nodes. A significant difference was superior viability of effector and suppressor cells from new onset diabetics in the presence of high interleukin-2 (IL-2) concentrations in vitro as compared to cells of prediabetic mice. Overall, we found no correlation between FoxP3(+) Treg in the pancreatic lymph nodes and the inflammatory scores of individual NOD mice. CD25(-)FoxP3(+) Treg are markedly increased in the pancreatic infiltrates in late stages of inflammation, possibly an effort to counteract destructive insulitis. Considering extensive evidence that Treg in aged NOD mice are functionally sufficient, quantitative profiling evolves as an unreliable tool to assess mechanism and causes of inflammation under baseline conditions. Immune profiles are modulated by thymic output, cell migration, shedding of markers, proliferation, survival and in-situ evolution of regulatory cells.


Downregulation of TFPI in breast cancer cells induces tyrosine phosphorylation signaling and increases metastatic growth by stimulating cell motility.
Increased hemostatic activity is common in many cancer types and often causes additional complications and even death. Circumstantial evidence suggests that tissue factor pathway inhibitor-1 (TFPI) plays a role in cancer development. We recently reported that downregulation of TFPI inhibited apoptosis in a breast cancer cell line. In this study, we investigated the effects of TFPI on self-sustained growth and motility of these cells, and of another invasive breast cancer cell type (MDA-MB-231). Stable cell lines with TFPI (both α and β) and only TFPIβ downregulated were created using RNA interference technology. We investigated the ability of the transduced cells to grow, when seeded at low densities, and to form colonies, along with metastatic characteristics such as adhesion, migration and invasion. Downregulation of TFPI was associated with increased self-sustained cell growth. An increase in cell attachment and spreading was observed to collagen type I, together with elevated levels of integrin α2. Downregulation of TFPI also stimulated migration and invasion of cells, and elevated MMP activity was involved in the increased invasion observed. Surprisingly, equivalent results were observed when TFPIβ was downregulated, revealing a novel function of this isoform in cancer metastasis. Our results suggest an anti-metastatic effect of TFPI and may provide a novel therapeutic approach in cancer.


Micelles-encapsulated microcapsules for sequential loading of hydrophobic and water-soluble drugs.
Layer-by-layer (LbL) assembly was conducted on CaCO(3) microparticles pre-doped with polystyrene-block-poly(acrylic acid) (PS-b-PAA) micelles, and resulted in micelles encapsulation in the microcapsules after core removal. Distribution of the micelles in the templates and capsules was characterized by transmission electron microscopy and confocal laser scanning microscopy. The micelles inside the capsules connected with each other to form a chain and network-like structure with a higher density near the capsule walls. The hydrophobic PS cores were then able to load small uncharged hydrophobic drugs while the negatively charged PAA corona could induce spontaneous deposition of water-soluble positively charged drugs such as doxorubicin.


The Child Outcome Rating Scale: validating a four-item measure of psychosocial functioning in community and clinic samples of children aged 10-15.
Psychosocial functioning is considered an important and valued outcome in relation to young people's mental health as a construct distinct from psychiatric symptomology, especially in the light of an increasing focus on transdiagnostic approaches. Yet, the level of psychosocial functioning is rarely directly asked of young people themselves, despite the widespread recognition that the young person's perspective is valuable and is often at odds with those of other reporters, such as parents or professionals. One possible reason for this is that the field lacks a clear agreed tool to capture this information in a non-burdensome way. To begin to address this gap, this paper describes psychometric analysis of the Child Outcome Rating Scale (CORS), a brief and highly accessible self-report measure of young people's psychosocial functioning already used extensively by mental health professionals around the world but with only limited data on psychometric robustness. Using large community (n = 7822) and clinic (n = 2604) samples, we explore the factor structure, construct validity, internal consistency, differential item functioning, and sensitivity of the CORS. We found that the CORS stands up to psychometric scrutiny, having found satisfactory levels of reliability, validity, and sensitivity in this sample. We also found that the CORS is suitable for use with young people as old as 15 years old. That the CORS has been found to be psychometrically robust while being highly feasible (brief, simple, easy to administer) for use in busy clinical settings, combined with the fact that the CORS has already been widely adopted by clinicians and young people, suggests CORS may be an important tool for international use.


Generation of plants resistant to tomato yellow leaf curl virus by using artificial zinc-finger proteins.
Previously, we designed an artificial zinc-finger protein (AZP) for blocking a replication protein (Rep) of beet severe curly top virus (BSCTV) from binding to its replication origin and demonstrated that transgenic Arabidopsis plants expressing the AZP are completely resistant to the virus infection. Here we applied the AZP technology to tomato yellow leaf curl virus (TYLCV) infective to an important agricultural crop, tomato. We designed and constructed an AZP binding to the direct repeat to block the TYLCV Rep binding. In gel shift assays, we confirmed that the designed AZP has a higher affinity to the replication origin than that of Rep and that the AZP effectively inhibited the Rep binding to its replication origin in vitro. The AZP gene was then introduced into a plant genome with the help of Agrobacterium tumefaciens to generate the transgenic plants. We will discuss properties of the AZP-transgenic plants against TYLCV infection.


Heading in Football: insights from stakeholders in amateur football.
Despite emerging research questioning the long-term effect of purposeful heading on players' brain health, heading-related perspectives and behaviours of stakeholders in amateur football in Australia (a country without heading guidelines) remain unknown. This study aimed to explore the current heading-related perspectives and behaviours of football stakeholders. In total, 290 players (aged over 11 years), 54 coaches, 34 non-coaching staff and 14 medical staff completed the survey. Of the 290 players, 56.5% reported being formally trained in heading, with female players less likely to be trained than male players (p < 0.05). Players were the least concerned about the long-term effects of heading, while medical staff were the most concerned (33.1% and 57.1%, respectively). From proposed strategies to reduce heading burden, a heading ban for all ages was least popular (2.3%), while teaching heading technique was most popular (67.3%). Our study provides insights into football stakeholders' heading-related perspectives, which could be used, along with scientific evidence, to inform pragmatic future heading guidelines.


Recent advances in understanding the mechanisms of cerebellar granule cell development and function and their contribution to behavior.
The cerebellum is the focus of an emergent series of debates because its circuitry is now thought to encode an unexpected level of functional diversity. The flexibility that is built into the cerebellar circuit allows it to participate not only in motor behaviors involving coordination, learning, and balance but also in non-motor behaviors such as cognition, emotion, and spatial navigation. In accordance with the cerebellum's diverse functional roles, when these circuits are altered because of disease or injury, the behavioral outcomes range from neurological conditions such as ataxia, dystonia, and tremor to neuropsychiatric conditions, including autism spectrum disorders, schizophrenia, and attention-deficit/hyperactivity disorder. Two major questions arise: what types of cells mediate these normal and abnormal processes, and how might they accomplish these seemingly disparate functions? The tiny but numerous cerebellar granule cells may hold answers to these questions. Here, we discuss recent advances in understanding how the granule cell lineage arises in the embryo and how a stem cell niche that replenishes granule cells influences wiring when the postnatal cerebellum is injured. We discuss how precisely coordinated developmental programs, gene expression patterns, and epigenetic mechanisms determine the formation of synapses that integrate multi-modal inputs onto single granule cells. These data lead us to consider how granule cell synaptic heterogeneity promotes sensorimotor and non-sensorimotor signals in behaving animals. We discuss evidence that granule cells use ultrafast neurotransmission that can operate at kilohertz frequencies. Together, these data inspire an emerging view for how granule cells contribute to the shaping of complex animal behaviors.


Comparing Prophylactic Effect of Levetiracetam, Sodium Valproate, and Propranolol in Pediatric Migraine: A Randomized Clinical Trial.
Epidemiologic studies point to an increased prevalence of migraine in children in recent decades. Migraine treatment involves acute and prophylactic therapy. Recently, such anti-epileptic drugs as Levetiracetam have been used to treat adult migraines. The present study aimed to compare the efficacy of Levetiracetam, Sodium Valproate, and Propranolol in preventing migraine headaches in children. In this clinical trial, children with migraine were randomly divided into three groups. Each group consisted of 13 children. Two groups were treated with Propranolol and Sodium Valproate, respectively. Another group (the case) was treated with Levetiracetam. The patients were assessed based on headache score, PedMIDAS, and headache frequency before and three months after the intervention. Finally, the data was analyzed using descriptive and analytical statistical methods. Levetiracetam significantly reduced the headache severity (P=0.026), frequency (P=0.024), and PedMIDAS score (P=0.001) in children with migraine. However, no significant difference was found between the three groups. The percentage of patients who experienced pain relief was detected as 69.24%, 92.31%, and 30.76% in the Propranolol, Sodium Valproate, and Levetiracetam groups, respectively. This study concluded that Levetiracetam can be used as a migraine prophylaxis drug in children.


[Innovative practices for health care].
The need to build a new look to the care, based on dialogue and creativity, enables the social transformation of the role of professionals in their practice. It is understood here that the care is more than an act: it is an attitude. Moreover, the conceptual framework of psychosocial rehabilitation is an instrument that might enable the construction of this new look. In this context, to care is to consider the importance of building projects of life, significant for each user, as lynchpin of the therapeutic action. The necessary changes in mental health's practice and psychiatric nursing are progressing, since the knowledge produced in this area incorporates strategies to welcome and maintenance which considers, at all times, the exercise of active citizenship of the mental patients.


Selective Extraction of Sinapic Acid Derivatives from Mustard Seed Meal by Acting on pH: Toward a High Antioxidant Activity Rich Extract.
The aim of this paper is to study the effect of the pH on the extraction of sinapic acid and its derivatives from mustard seed meal. Solutions of acidic pH (pH 2), basic pH (pH 12) and distilled water (uncontrolled pH ~ 4.5) were tested at different percentages of ethanol. The maximum extraction yield for sinapic acid (13.22 µmol/g of dry matter (DM)) was obtained with a buffered aqueous solution at pH 12. For ethyl sinapate, the maximum extraction yield reached 9.81 µmol/g DM with 70% ethanol/buffered aqueous solution at pH 12. The maximum extraction yield of sinapine (15.73 µmol/g DM) was achieved with 70% ethanol/buffered aqueous solution at pH 2. The antioxidant activity of each extract was assessed by DPPH assay; the results indicated that the extracts obtained at pH 12 and at low ethanol percentages (<50%) exhibit a higher antioxidant activity than extracts obtained at acidic conditions. Maximum antioxidant activity was reached at pH 12 with buffer solution (11.37 mg of Trolox Equivalent/g DM), which confirms that sinapic acid-rich fractions exhibit a higher antioxidant activity. Thus, to obtain rich antioxidant extracts, it is suggested to promote the presence of sinapic acid in the extracts.


Lymphocytes from ponies experimentally infected with equine herpesvirus 1: subpopulation dynamics and their response to mitogens.
Six pony foals, free of detectable serum neutralization (SN) antibody against equine herpesvirus type 1 by the standard virus-neutralization (VN) test, were inoculated with equine herpesvirus type 1. The ponies showed typical clinical signs of respiratory tract disease and developed a transient leukopenia, involving lymphocytes as well as neutrophils. The leukopenia reached its lowest point on postinoculation days (PID) 3 to 5 and then returned to base-line values by PID 8 to 10. On quantitation of lymphocyte subpopulations, T and B lymphocytes were decreased during the onset of leukopenia and then recouped during the recovery from leukopenia. However, the proportions of the T and B lymphocytes remained constant during the lymphopenia, ranging from 70% to 80% and 20% to 30%, respectively. The lymphocyte blastogenic response to mitogens increased to peak by PID 2 to 5 and then decreased to base line values or below by PID 7. Mitogen responses of T lymphocyte and mixed lymphocyte preparations were nearly similar. However, the responses of 2 ponies wee somewhat different from the responses of others in that there was an increase in the B lymphocytes in the range of 40% to 50% during the recovery phase of lymphopenia. Also, the 2 ponies' mixed lymphocyte response to mitogens was considerably higher and the T lymphocyte response to mitogen was lower as compared with that of mixed lymphocyte preparation. The importance of these 2 types of responses is discussed.


A pilot study to assess cognition and pillbox fill accuracy by community-dwelling older adults.
To assess pillbox fill accuracy and cognition among community-dwelling older adults. A descriptive, cross-sectional study. Retail pharmacy. Convenience sample of English-speaking adults older than 60 years of age without dementia, taking more than four medications, and naive to Mediset use. In face-to-face interviews, subjects provided demographic, medical, and medication information, completed the Mini-Cog and Medi-Cog (combination of Mini-Cog and medication-transfer screen [MTS]), and filled their own medications in a pillbox. Data were analyzed using descriptive statistics and stepwise regression analysis with correctly filled pill count (PC) as the dependent variable and the cognitive screens as independent variables. Accuracy of the Mini-Cog, MTS, and Medi-Cog in predicting PC. Among 50 subjects (58% female, mean age 76.4 years), only one subject failed to pass the Mini-Cog and two failed to reach the criterion level of correctly filled PC. The mean (standard deviation) Mini-Cog score for the sample was 4.38 (0.81), MTS score was 4.1 (1.31), Medi-Cog score was 8.48 (1.82), and the mean PC was 97% (8%). The Mini-Cog and MTS individually accounted for about 30% of the variance (P < 0.001); the Medi-Cog accounted for 44% of the variance (P < 0.001), indicating strongest PC prediction. Nearly all study participants filled pillboxes accurately. The Medi-Cog was the strongest predictor of pillbox fill accuracy. Future studies of medication self-management abilities among community-dwelling older adults should include representative samples of this population, comprehensive assessment of health status, cognitive screening, pillbox fill accuracy, and the utilization of medications in filled pillboxes.


Inhibition of crotoxin phospholipase A(2) activity by manoalide associated with inactivation of crotoxin toxicity and dissociation of the heterodimeric neurotoxic complex.
Crotoxin (CACB complex) is a convulsant heterodimeric neurotoxic phospholipase A(2) (PLA(2)). The role of phospholipid hydrolysis in its epileptogenic properties remains unresolved. We, thus, studied the effect of manoalide (MLD), a PLA(2) inhibitor, on the toxin catalytic activity and its central and peripheral toxicity. Incubation of crotoxin with MLD fully and irreversibly inactivated its enzymatic activity. Interestingly, crotoxin also lost its central neurotoxicity after intracerebroventricular injection and peripheral toxicity after intravenous administration. MLD-treated crotoxin prevented the high affinity binding of [125I]-radiolabeled crotoxin on rat cortex synaptic plasma membranes. Further analysis of MLD-treated crotoxin by non-denaturing PAGE and surface plasmon resonance indicated that the crotoxin complex was dissociated after MLD treatment. Although the loss of MLD-treated crotoxin peripheral neurotoxicity could not be attributed to this dissociation, the presence of free CA subunit might explain the observed competition in binding experiments. In conclusion, the dissociation of the crotoxin complex by MLD, as demonstrated in this study, did not permit to specify the role of the enzymatic activity in crotoxin epileptogenic properties. Other approaches would be required to resolve this question.


Online self-study of chest X-rays shows no difference between blocked and mixed practice.
Chest radiograph interpretation is a complex skill and learners may benefit from deliberate instructional design modalities, such as mixed practice. Proposed benefits of mixed over blocked practice include the elimination of cueing and the highlighting of contrasting features. However, current evidence for the superiority of mixed practice is conflicting. This study compares mixed versus blocked practice, after the initial teaching of concepts, among medical students using online self-study chest X-ray (CXR) modules. Two online CXR modules were developed that cover identical content but differ in the organisation of practice images. Blocked modules provided practice CXRs after each category, whereas mixed modules randomly ordered practice radiographs after all categories had been taught. Medical students in Years 1-3 were randomised to either module and were tested on 20 new CXRs immediately after completion and at 2 weeks. The primary outcome was immediate diagnostic accuracy. Secondary outcomes included diagnostic accuracy at 2 weeks, time per module and reported module difficulty. A total of 58 medical students participated (32 in the blocked and 26 in the mixed module). Level of training and previous CXR experience were similar across the groups. Totals of 1160 and 1120 answers were evaluated for immediate and 2 week post-test scores, respectively. There were no significant differences in mean diagnostic accuracy between the blocked (mean score: 11.7/20) and mixed (mean score: 11.0/20) practice groups on immediate testing (t = 0.83, d.f. = 56, p = 0.41) or at 2 weeks (mean score: 11.2/20 versus 10.9/20; t = 0.518, d.f. = 54, p = 0.61). Post-test scores showed no correlation with training level (R = 0.23, p = 0.09) or completion time (R = -0.09, p = 0.5). Reported module difficulty was similar between the mixed (3.22/5) and blocked (3.19/5) groups. On multivariable linear regression controlling for completion time, training level and CXR experience, between-group differences remained non-significant. Performance after mixed practice was similar to that after blocked practice. Results may reflect similarities between modules in teaching, which emphasised contrast learning, greater effect of initial teaching rather than practice, or absence of tutor-led instruction. Alternatively, results may reflect the higher cognitive load in mixed practice imposed by contrasting multiple diagnoses.


Amplitude-based height-reconstruction techniques for synthetic aperture ladar systems.
We examine the performance of amplitude-based height-estimation techniques for use with airborne synthetic aperture ladar (SAL) sensors in generating three-dimensional reconstructions of ground targets. Such techniques lend themselves to implementation more readily than phase-based techniques and are also more tolerant to phase instabilities that might be associated with SAL systems. For pairwise amplitude-comparison monopulse processing, we present analyses of the expected height sensitivity and bias of SAL systems in terms of the system parameters. We verify this analysis with simulations, and we also provide an overview of other SAL phenomena that affect height-estimation accuracy. We then propose an array-based joint-processing approach that can be applied instead of pairwise monopulse processing. We show that the joint-processing approach represents the maximum-likelihood estimator for obtaining the target height, and we demonstrate that the proposed approach significantly reduces bias-induced errors.


Identification of novel genes in the oral pathogen Campylobacter rectus.
A poorly described bacterium, Campylobacter rectus, has been implicated as an etiological agent of periodontal disease. The aim of this study was to use a comparative genomics approach to identify genes that contribute to the lifestyle of C. rectus as an oral pathogen. Suppressive subtractive hybridization was used to identify genes encoded by C. rectus ATCC 33238, but not present in the genome of a related Campylobacter species, Campylobacter jejuni ATCC 11168. Suppressive subtractive hybridization identified 154 unique DNA sequences from the C. rectus genome. Ninety-two of the 154 clones were classified as C. rectus-specific, as they did not show significant sequence homology to genes identified in any strain of C. jejuni (blast E-value >1E-3). blast analysis predicted that the 92 C. rectus-specific gene fragments play a role in a variety of biological processes including signal transduction mechanisms (histidine kinase, response regulators, diguanylate cyclases, chemotaxis receptor) and potentially virulence (S-layer RTX and cysteine desulfhydrase). Further analysis of the C. rectus-specific clones showed that 10 genes had Campylobacter homologues that were only found in species that commonly reside within the oral cavity of humans and 10 other fragments shared homology only with non-campylobacter organisms. These data provide the first substantial insights into the genomic content of C. rectus, a significant oral pathogen. The genes identified in this study are a valuable resource for initiating new research on the virulence of C. rectus during periodontitis.


The acute effects of daily nicotine intake on heart rate--a toxicokinetic and toxicodynamic modelling study.
Joint physiologically-based toxicokinetic and toxicodynamic (PBTK/TD) modelling was applied to simulate concentration-time profiles of nicotine, a well-known stimulant, in the human body following single and repeated dosing. Both kinetic and dynamic models were first calibrated by using in vivo literature data for the Caucasian population. The models were then used to estimate the blood and liver concentrations of nicotine in terms of the Area Under Curve (AUC) and the peak concentration (Cmax) for selected exposure scenarios based on inhalation (cigarette smoking), oral intake (nicotine lozenges) and dermal absorption (nicotine patches). The model simulations indicated that whereas frequent cigarette smoking gives rise to high AUC and Cmax in blood, the use of nicotine-rich dermal patches leads to high AUC and Cmax in the liver. Venous blood concentrations were used to estimate one of the most common acute effects, mean heart rate, both at rest and during exercise. These estimations showed that cigarette smoking causes a high peak heart rate, whereas dermal absorption causes a high mean heart rate over 48h. This study illustrates the potential of using PBTK/TD modelling in the safety assessment of nicotine-containing products.


Green light induces transcription of the phycoerythrin operon in the cyanobacterium Calothrix 7601.
Phycobilisomes, the major light-harvesting complexes of cyanobacteria are multimolecular structures made up of chromophoric proteins called phycobiliproteins and non chromophoric linker polypeptides. We report here the isolation and nucleotide sequence of the genes, cpeA and cpeB, which in Calothrix PCC 7601 encode the alpha and beta subunits of phycoerythrin, one of the major phycobiliproteins. In Calothrix PCC 7601, modulation of the polypeptide composition of the phycobilisomes occurs in response to changes of the light wavelength, a phenomenon known as complementary chromatic adaptation. Under green illumination, cells synthesize phycoerythrin and its two specifically associated linker polypeptides (LR35 and LR36), while under red illumination none of these proteins are detected. Using specific probes, a single transcript (1450 nucleotide long) corresponding to the cpe genes was detected but only in green-light-grown cells, establishing the occurrence of transcriptional regulation for the expression of this operon in response to light wavelength changes. The size of this transcript excludes the possibility that the phycoerythrin-associated LR35 and LR36 could be cotranscribed with the cpeA and cpeB genes.


[Non-VKA oral anticoagulants: an update for the clinical biologists].
Non-VKA oral anticoagulants (NOACs), thanks to their ease of use and their similar or superior safety/efficacy profiles versus warfarin, have now widely reached the lucrative market of anticoagulation. However, while the marketing authorization holders always claim, in a quite unclear way that no monitoring is required, accumulative evidence and cases of major bleeding have been described in the literature and reported by spontaneous reporting systems at the regulator's level. These compounds are usually given at fixed doses without routine coagulation monitoring. However, new data suggests that an assessment of the response at the individual level could improve the benefit-risk ratio of, at least dabigatran. Therefore, in certain patient populations, i.e. acute or chronic renal impairment or multiple drug interactions, measurement of drug exposure may be useful to ensure an optimal treatment response. More specific circumstances such as patients experiencing a haemorrhagic or thromboembolic event during the treatment duration, patients who require urgent surgery or an invasive procedure, or patient with a suspected overdose could benefit from such a measurement. This article aims at providing guidance on how to best estimate the intensity of anticoagulation using laboratory assays in daily practice.


[Diagnostic problems in dizziness or vertigo (author's transl)].
Different causes of dizziness or vertigo can only be recognized by thorough anamnestic explorations. Following a classification in vestibular and nonvestibular causes for vertigo, a further differentiation is possible by defining different characteristic qualities of the symptoms involved. In addition to the classical vestibular forms of vertigo seen, dizziness currently results from drug overdosages, hypertension, polyneuropathy and--less commonly, but equally important--brief epileptic seizures. Psychosomatic and neurotic symptoms may also lead to unsteady gait, dizziness or vertigo, all of which are distinguished only with difficulty by the patient.


Designing Molecular Building Blocks for the Self-assembly of Complex Porous Networks.
The creation of molecular or colloidal building blocks which can self-assemble into complex, ordered porous structures has been long sought-after, and so are the guiding principles behind this creation. The pursuit of this goal has led to the creation of novel classes of materials like metal organic frameworks (MOFs) and covalent organic frameworks (COFs). In theory, a tremendous number of structures can be formed by these materials due to the variety of geometries available to their building blocks. However, most realized crystal structures tend to be simple or homoporous and typically assemble from building blocks with high degrees of symmetry. Building blocks with low degrees of symmetry suitable for assembly into the more complex structures tend to assemble into polymorphous or disordered structures instead. In this work, we use Monte Carlo simulations of patchy vertex-like building blocks to show how the addition of chemical specificity via orthogonally reacting functional sites can allow vertex-like building blocks with even asymmetric geometries to self-assemble into ordered crystallites of various complex structures. In addition to demonstrating the utility of such a strategy in creating ordered, heteroporous structures, we also demonstrate that it can be used as a means for tuning specific features of the crystal structure, accomplishing such aims as the control of relative pore sizes. We also discuss heuristics for properly designing molecules so that they can assemble into target structures.


Doctor Recommendation Model Based on Ontology Characteristics and Disease Text Mining Perspective.
Patients can access medical services such as disease diagnosis online, medical treatment guidance, and medication guidance that are provided by doctors from all over the country at home. Due to the complexity of scenarios applying medical services online and the necessity of professionalism of knowledge, the traditional recommendation methods in the medical field are confronting with problems such as low computational efficiency and poor effectiveness. At the same time, patients consulting online come from all sides, and most of them suffer from nonacute or malignant diseases, and hence, there may be offline medical treatment. Therefore, this paper proposes an online prediagnosis doctor recommendation model by integrating ontology characteristics and disease text. Particularly, this recommendation model takes full consideration of geographical location of patients. The recommendation model takes the real consultation data from online as the research object, fully testifying its effectiveness. Specifically, this model would make recommendation to patients on department and doctors based on patients' information of symptoms, diagnosis, and geographical location, as well as doctor's specialty and their department. Utilizing crawler technique, five hospital departments were selected from the online medical service platform. The names of the departments were in accordance with the standardized department names used in real hospitals (e.g., endocrinology, dermatology, gynemetrics, pediatrics, and neurology). As a result, a dataset consisting of 20000 consultation questions by patients was built. Through the application of Python and MySQL algorithms, replacing semantic dictionary retrieval or word frequency statistics, word vectors were utilized to measure similarity between patients' prediagnosis and doctors' specialty, forming a recommendation framework on medical departments or doctors based on the above-obtained sentence similarity measurement and providing recommendation advices on intentional departments and doctors. In the online medical field, compared with the traditional recommendation method, the model proposed in the paper is of higher recommendation accuracy and feasibility in terms of department and doctor recommendation effectiveness. The proposed online prediagnosis doctor recommendation model integrates ontology characteristics and disease text mining. The model gives a relatively more accurate recommendation advice based on ontology characteristics such as patients' description texts and doctors' specialties. Furthermore, the model also gives full consideration on patients' location factors. As a result, the proposed online prediagnosis doctor recommendation model would improve patients' online consultation experience and offline treatment convenience, enriching the value of online prediagnosis data.


Clusterin and Its Role in Insulin Resistance and the Cardiometabolic Syndrome.
The cardiometabolic syndrome involves a clustering of metabolic and cardiovascular factors which increase the risk of patients developing both Type 2 Diabetes Mellitus and cardio/cerebrovascular disease. Although the mechanistic underpinnings of this link remain uncertain, key factors include insulin resistance, excess visceral adiposity, atherogenic dyslipidemia, and endothelial dysfunction. Of these, a state of resistance to insulin action in overweight/obese patients appears to be central to the pathophysiologic process. Given the increasing prevalence of obesity-related Type 2 Diabetes, coupled with the fact that cardiovascular disease is the number one cause of mortality in this patient population, a more thorough understanding of the cardiometabolic syndrome and potential options to mitigate its risk is imperative. Inherent in the pathogenesis of insulin resistance is an underlying state of chronic inflammation, at least partly in response to excess adiposity. Within obese adipose tissue, an immunomodulatory shift occurs, involving a preponderance of pro-inflammatory immune cells and cytokines/adipokines, along with antigen presentation by adipocytes. Therefore, various adipokines differentially expressed by obese adipocytes may have a significant effect on cardiometabolism. Clusterin is a molecular chaperone that is widely produced by many tissues throughout the body, but is also preferentially overexpressed by obese compared lean adipocytes and relates strongly to multiple components of the cardiometabolic syndrome. Herein, we summarize the known and potential roles of circulating and adipocyte-specific clusterin in cardiometabolism and discuss potential further investigations to determine if clusterin is a viable target to attenuate both metabolic and cardiovascular disease.


Iron, Zinc and Phytic Acid Retention of Biofortified, Low Phytic Acid, and Conventional Bean Varieties When Preparing Common Household Recipes.
Biofortification is an effective method to improve the nutritional content of crops and nutritional intake. Breeding for higher micronutrient mineral content in beans is correlated with an increase in phytic acid, a main inhibitor of mineral absorption in humans. Low phytic acid (lpa) beans have a 90% lower phytic acid content compared to conventional beans. This is the first study to investigate mineral and total phytic acid retention after preparing common household recipes from conventional, biofortified and lpa beans. Mineral retention was determined for two conventional, three biofortified and two lpa bean genotypes. Treatments included soaking, boiling (boiled beans) and refrying (bean paste). The average true retention of iron after boiling was 77.2-91.3%; for zinc 41.2-84.0%; and for phytic acid 49.9-85.9%. Soaking led to a significant decrease in zinc and total phytic acid after boiling and refrying, whereas for iron no significant differences were found. lpa beans did not exhibit a consistent pattern of difference in iron and phytic acid retention compared to the other groups of beans. However, lpa beans had a significantly lower retention of zinc compared to conventional and biofortified varieties (p < 0.05). More research is needed to understand the underlying factors responsible for the differences in retention between the groups of beans, especially the low retention of zinc. Combining the lpa and biofortification traits could further improve the nutritional benefits of biofortified beans, by decreasing the phytic acid:iron and zinc ratio in beans.


Longitudinal brain white matter alterations in minimal hepatic encephalopathy before and after liver transplantation.
Cerebral edema is the common pathogenic mechanism for cognitive impairment in minimal hepatic encephalopathy. Whether complete reversibility of brain edema, cognitive deficits, and their associated imaging can be achieved after liver transplantation remains an open question. To characterize white matter integrity before and after liver transplantation in patients with minimal hepatic encephalopathy, multiple diffusivity indices acquired via diffusion tensor imaging was applied. Twenty-eight patients and thirty age- and sex-matched healthy volunteers were included. Multiple diffusivity indices were obtained from diffusion tensor images, including mean diffusivity, fractional anisotropy, axial diffusivity and radial diffusivity. The assessment was repeated 6-12 month after transplantation. Differences in white matter integrity between groups, as well as longitudinal changes, were evaluated using tract-based spatial statistical analysis. Correlation analyses were performed to identify first scan before transplantation and interval changes among the neuropsychiatric tests, clinical laboratory tests, and diffusion tensor imaging indices. After transplantation, decreased water diffusivity without fractional anisotropy change indicating reversible cerebral edema was found in the left anterior cingulate, claustrum, postcentral gyrus, and right corpus callosum. However, a progressive decrease in fractional anisotropy and an increase in radial diffusivity suggesting demyelination were noted in temporal lobe. Improved pre-transplantation albumin levels and interval changes were associated with better recoveries of diffusion tensor imaging indices. Improvements in interval diffusion tensor imaging indices in the right postcentral gyrus were correlated with visuospatial function score correction. In conclusion, longitudinal voxel-wise analysis of multiple diffusion tensor imaging indices demonstrated different white matter changes in minimal hepatic encephalopathy patients. Transplantation improved extracellular cerebral edema and the results of associated cognition tests. However, white matter demyelination may advance in temporal lobe.


Phosphorylation of adenovirus DNA-binding protein.
Evidence is presented here which indicates that the adenovirus DNA-binding protein (DBP) is phosphorylated at a tyrosine residue early in infection. This was suggested by the discovery that a proportion of the label in 32P-labelled DBP was resistant to alkali, and was substantiated by acid hydrolysis of DBP immunoprecipitates and by immunoblotting with a monoclonal antibody against phosphotyrosine. Treatment of [35S]methionine-labelled DBPs with chymotrypsin produced fragments of apparent Mr 45K and 39K whereas digestion of 32P-labelled DBP resulted in fragments of 45K and 26K. Consideration of the distribution of 32P label and its alkali stability in these fragments suggested that chymotrypsin cleaved populations of DBP at different sites depending on their phosphorylation states. The conservation, in all of the seven adenovirus serotypes sequenced, of a tyrosine residue (at amino acid 195 in adenovirus type 2) together with its surrounding residues, suggests that phosphorylation/dephosphorylation at this tyrosine residue may be important in various functions ascribed to the DBP.


Rehabilitation of executive functioning in patients with frontal lobe brain damage with goal management training.
Executive functioning deficits due to brain disease affecting frontal lobe functions cause significant real-life disability, yet solid evidence in support of executive functioning interventions is lacking. Goal Management Training (GMT), an executive functioning intervention that draws upon theories concerning goal processing and sustained attention, has received empirical support in studies of patients with traumatic brain injury, normal aging, and case studies. GMT promotes a mindful approach to complex real-life tasks that pose problems for patients with executive functioning deficits, with a main goal of periodically stopping ongoing behavior to monitor and adjust goals. In this controlled trial, an expanded version of GMT was compared to an alternative intervention, Brain Health Workshop that was matched to GMT on non-specific characteristics that can affect intervention outcome. Participants included 19 individuals in the chronic phase of recovery from brain disease (predominantly stroke) affecting frontal lobe function. Outcome data indicated specific effects of GMT on the Sustained Attention to Response Task as well as the Tower Test, a visuospatial problem-solving measure that reflected far transfer of training effects. There were no significant effects on self-report questionnaires, likely owing to the complexity of these measures in this heterogeneous patient sample. Overall, these data support the efficacy of GMT in the rehabilitation of executive functioning deficits.


Estrous cycle plasticity in the hyperpolarization-activated current ih is mediated by circulating 17β-estradiol in preoptic area kisspeptin neurons.
Circulating gonadal steroid hormones are thought to modulate a wide range of brain functions. However, the effects of steroid fluctuations through the ovarian cycle on the intrinsic properties of neurons are not well understood. We examined here whether gonadal steroids modulated the excitability of kisspeptin neurons located in the rostral periventricular region of the third ventricle (RP3V) of female mice. These cells are strongly implicated in sensing the high levels of circulating estradiol on proestrus to activate gonadotropin-releasing hormone (GnRH) neurons that, in turn, trigger ovulation. Electrophysiological studies were undertaken in brain slices from ovariectomized (OVX), diestrous, and proestrous kisspeptin-GFP mice. RP3V kisspeptin neurons exhibited marked changes in the hyperpolarization-evoked depolarizing sag and rebound firing across these groups. The hyperpolarization-activated current Ih was identified to be responsible for the depolarizing sag and was increased across OVX → diestrous → proestrous mice. Experiments in OVX mice given estradiol replacement identified an estradiol-dependent increase in Ih within RP3V kisspeptin neurons. Ih in these cells was found to contribute to their subthreshold membrane properties and the dynamics of rebound firing following hyperpolarizing stimuli in an estrous cycle-dependent manner. Only a minor role was found for Ih in modulating the spontaneous burst firing of RP3V kisspeptin neurons. These observations identify Ih as an ionic current that is regulated in a cyclical manner by circulating estradiol within the female brain, and suggest that such plasticity in the intrinsic properties of RP3V kisspeptin neurons may contribute to the generation of the preovulatory GnRH surge.


A synthetic approach to kingianin A based on biosynthetic speculation.
A synthetic approach towards the structurally complex dimer, kingianin A is reported. The strategy involved a cascade of complexity generating reactions, inspired through biosynthetic speculation. A concise protecting group free synthesis of the proposed monomeric precursor pre-kingianin A has been achieved using a tandem Stille cross-coupling reaction and electrocyclisation process. However, preliminary studies of the key dimerisation reaction have been conducted, which indicate that the process is not spontaneous, raising questions as to the origin of this complex natural product.


Depletion of Intense Fields.
The interaction of charged particles and photons with intense electromagnetic fields gives rise to multiphoton Compton and Breit-Wheeler processes. These are usually described in the framework of the external field approximation, where the electromagnetic field is assumed to have infinite energy. However, the multiphoton nature of these processes implies the absorption of a significant number of photons, which scales as the external field amplitude cubed. As a result, the interaction of a highly charged electron bunch with an intense laser pulse can lead to significant depletion of the laser pulse energy, thus rendering the external field approximation invalid. We provide relevant estimates for this depletion and find it to become important in the interaction between fields of amplitude a_{0}∼10^{3} and electron bunches with charges of the order of 10 nC.


Nonstarch polysaccharides modulate bacterial microbiota, pathways for butyrate production, and abundance of pathogenic Escherichia coli in the pig gastrointestinal tract.
The impact of nonstarch polysaccharides (NSP) differing in their functional properties on intestinal bacterial community composition, prevalence of butyrate production pathway genes, and occurrence of Escherichia coli virulence factors was studied for eight ileum-cannulated growing pigs by use of terminal restriction fragment length polymorphism (TRFLP) and quantitative PCR. A cornstarch- and casein-based diet was supplemented with low-viscosity, low-fermentability cellulose (CEL), with high-viscosity, low-fermentability carboxymethylcellulose (CMC), with low-viscosity, high-fermentability oat beta-glucan (LG), and with high-viscosity, high-fermentability oat beta-glucan (HG). Only minor effects of NSP fractions on the ileal bacterial community were observed, but NSP clearly changed the digestion in the small intestine. Compared to what was observed for CMC, more fermentable substrate was transferred into the large intestine with CEL, LG, and HG, resulting in higher levels of postileal dry-matter disappearance. Linear discriminant analysis of NSP and TRFLP profiles and 16S rRNA gene copy numbers for major bacterial groups revealed that CMC resulted in a distinctive bacterial community in comparison to the other NSP, which was characterized by higher gene copy numbers for total bacteria, Bacteroides-Prevotella-Porphyromonas, Clostridium cluster XIVa, and Enterobacteriaceae and increased prevalences of E. coli virulence factors in feces. The numbers of butyryl-coenzyme A (CoA) CoA transferase gene copies were higher than those of butyrate kinase gene copies in feces, and these quantities were affected by NSP. The present results suggest that the NSP fractions clearly and distinctly affected the taxonomic composition and metabolic features of the fecal microbiota. However, the effects were more linked to the individual NSP and to their effect on nutrient flow into the large intestine than to their shared functional properties.


In Vitro and In Vivo Antibacterial Activity, Toxicity and Resistance Analysis of Pleuromutilin Derivative Z33 against Methicillin-Resistant Staphylococcus aureus.
The novel pleuromutilin derivative, which showed excellent in vitro antibacterial activity against MRSA, 22-(2-(2-(4-((4-(4-nitrophenyl)piperazin-1-yl)methyl)-1H-1,2,3-triazol-1-yl)acetamido)phenyl)thioacety-l-yl-22-deoxypleuromutilin (Z33), was synthesized and characterized in our previous work. In this study, the preliminary pharmacodynamics and safety of Z33 were further evaluated. In in vitro antibacterial activity assays, Z33 was found to be a potent bactericidal antibiotic against MRSA that induced dose-dependent growth inhibition and long-term post-antibiotic effect (PAE). The drug-resistance test demonstrated that Z33 possessed a narrow mutant selection window and lower propensities to select resistance than that of tiamulin. Cytochrome P450 (CYP450) inhibition assay determined that the inhibitory effect of Z33 was similar to that of tiamulin against the activity of CYP3A4, and was lower than that of tiamulin on the activity of CYP2E1. Toxicity determination showed that both Z33 and tiamulin displayed low cytotoxicity of RAW264.7 cells. Furthermore, Z33 was found to be a high-security compound with a 50% lethal dose (LD50) above 5000 mg/kg in the acute oral toxicity test in mice. In an in vivo antibacterial activity test, Z33 displayed better therapeutic effectiveness than tiamulin in the neutropenic mouse thigh infection model. In summary, Z33 was worthy of further development as a highly effective and safe antibiotic agent against MRSA infection.


Total variance, an estimator of long-term frequency stability [standards].
Total variance is a statistical tool developed for improved estimates of frequency stability at averaging times up to one-half the test duration. As a descriptive statistic, total variance performs an exact decomposition of the sample variance of the frequency residuals into components associated with increasing averaging times. As an estimator of Allan variance, total variance has greater equivalent degrees of freedom and lesser mean square error than the standard unbiased estimator has.


Multicoil resonance-based parallel array for smart wireless power delivery.
This paper presents a novel resonance-based multicoil structure as a smart power surface to wirelessly power up apparatus like mobile, animal headstage, implanted devices, etc. The proposed powering system is based on a 4-coil resonance-based inductive link, the resonance coil of which is formed by an array of several paralleled coils as a smart power transmitter. The power transmitter employs simple circuit connections and includes only one power driver circuit per multicoil resonance-based array, which enables higher power transfer efficiency and power delivery to the load. The power transmitted by the driver circuit is proportional to the load seen by the individual coil in the array. Thus, the transmitted power scales with respect to the load of the electric/electronic system to power up, and does not divide equally over every parallel coils that form the array. Instead, only the loaded coils of the parallel array transmit significant part of total transmitted power to the receiver. Such adaptive behavior enables superior power, size and cost efficiency then other solutions since it does not need to use complex detection circuitry to find the location of the load. The performance of the proposed structure is verified by measurement results. Natural load detection and covering 4 times bigger area than conventional topologies with a power transfer efficiency of 55% are the novelties of presented paper.


A new Epstein-Barr virus negative Burkitt's lymphoma derived cell-line. I. Analysis of cell surface markers and abnormal expression of HLA antigens.
CHEV, a new Epstein-Barr virus negative Burkitt's lymphoma derived cell line has been studied. Karyotype analyses demonstrated the t (8; 14) characteristic translocation. Cell surface characterisation of this line showed the presence of mu and chi immunoglobulin chains and beta 2-microglobulin and the absence of the complement receptor. We are unable to detect HLA--A, B alloantigens on the cell surface which was in contrast with an apparently normal expression of these antigens in the cytoplasm.


Dimerisation of myomesin: implications for the structure of the sarcomeric M-band.
The sarcomeric M-band is thought to provide a link between the thick and the elastic filament systems. So far, relatively little is known about its structural components and their three-dimensional organisation. Myomesin seems to be an essential component of the M-band, since it is expressed in all types of vertebrate striated muscle fibres investigated and can be found in its mature localisation pattern as soon as the first myofibrils are assembled. Previous work has shown that the N-terminal and central part of myomesin harbour binding sites for myosin, titin and muscle creatine kinase. Intrigued by the highly conserved domain layout of the C-terminal half, we screened for new interaction partners by yeast two-hybrid analysis. This revealed a strong interaction of myomesin with itself. This finding was confirmed by several biochemical assays. Our data suggest that myomesin can form antiparallel dimers via a binding site residing in its C-terminal domain 13. We suggest that, similar to alpha-actinin in the Z-disc, the myomesin dimers cross-link the contractile filaments in the M-band. The new and the already previously identified myomesin interaction sites are integrated into the first three-dimensional model of the sarcomeric M-band on a molecular basis.


Increasing the uptake of long-acting and permanent methods of family planning: A qualitative study with village midwives in East Java and Nusa Tenggara Barat Provinces, Indonesia.
this analysis aims to explore midwives' insights into the provision of long-acting and permanent methods of contraception (LAPMs) in the selected areas of East Java and Nusa Tenggara Barat (NTB) Provinces, Indonesia. a qualitative study using in-depth interviews was conducted with 12 village midwives from 12 villages, to explore their perceptions and experiences in delivering family planning services. the study was carried out in May-June 2013, as part of the baseline assessment in the Improving Contraceptive Method Mix (ICMM) study. We interviewed 12 village midwives working in 12 villages in six study districts: Tuban, Kediri, and Lumajang Districts in East Java Province; and Lombok Barat, Lombok Timur, and Sumbawa Districts in NTB Province. an interview guideline was used in all interviews. It covered several topics, such as community perceptions of LAPMs, availability of contraception and related equipment, availability of human resources, and midwives' efforts to improve LAPM coverage. All interviews were recorded and transcribed. Content and thematic analyses were carried out by grouping and coding the information based on the identified themes and topics. according to village midwives interviewed in this study, community-level acceptance of LAPMs has increased over time; however, some still prefer using short-acting methods for a long period. The reasons include lack of awareness about the benefits and side effects of LAPMs, fear of surgical procedures, rumored consequences (for example, that LAPMs would limit women's ability to perform hard physical labor), and religious beliefs. There were several challenges reported by village midwives in delivering LAPM services, such as confusion about midwives' eligibility to provide LAPM services, lack of Contraceptive Technology Update (CTU) and counseling trainings, and shortage of supporting equipment (such as exam tables and IUD and implant insertion kits). There were several strategies implemented by village midwives to improve LAPM use, including strengthening the counseling services, accompanying clients to higher health facilities to obtain LAPM services, and providing services for groups of clients. All village midwives emphasized the importance of strengthening collaboration among stakeholders to increase the uptake of LAPM services. as midwives are the main family planning providers in Indonesia, efforts to address their challenges is essential. Enabling a supportive policy environment, strengthening promotional activities, increasing the number of training programs designed for village midwives-in addition to enhancing inter-sectoral collaboration-are some recommendations to improve LAPM uptake in study areas.


Prevalence of and factors influencing smoking among medical and non-medical students in Tbilisi, Georgia.
Smoking is a serious problem that has a devastating impact on health. The objective of this study was to describe the prevalence of and factors influencing smoking among medical and non-medical students in Tbilisi, Georgia, as well as to determine whether medical education has an impact on smoking. A cross-sectional study was carried out at Tbilisi State Medical University and Tbilisi State University, both of which are located in Tbilisi, Georgia. A total of 400 4th-year students (200 students at each university) were asked to complete standardized questionnaires. Of the sample as a whole, 48.75% were identified as smokers and 51.25% were identified as nonsmokers. The mean age was 20.24 years among smokers and 20.26 years among nonsmokers. Of the medical students, 49.5% were smokers, as were 48.0% of the non-medical students. The male-to-female ratio in the study population was 0.9:1.1. Smoking was found to have a strong relationship with gender, males accounting for 65% of all smokers. Of the smokers, 56.9% stated that they would like to quit smoking (for health or financial reasons). Of the medical students, 59.5% expressed a willingness to quit smoking, as did 54.2% of the non-medical students. There is a need to improve smoking education for undergraduate students. Special attention should be given to the inclusion of anti-smoking education in undergraduate curricula, as well as to the implementation of smoking prevention campaigns at institutions of higher education. However, such measures will be effective only if tobacco control policies are strictly enforced on the national level as well.


Pacific Islanders--migration and health.
Native Hawaiians and peoples from American Samoa, Guam and the Trust Territories of the Pacific Islands are all recipients of US subsidized health care. Categorized as Pacific Islanders they are a heterogeneous group with differences in biology, cultural adaptation to varied ecological settings, historical influences resulting from colonialism and present-day political factionalism. Yet, westernization on home islands and migration to Hawaii and the western United States have created similarities in disease patterns among these culturally diverse peoples. They have high rates of the chronic diseases of civilization: cardiovascular disease, diabetes mellitus and hypertension. Obesity, associated with these ailments, has become a major health problem among Pacific Islanders and may be attributed to changes in local food production and consumption in conjunction with sedentarization. Culturally and linguistically distinct from the American mainstream, these people as migrants or residents are marginal within the US social structure and find if difficult to obtain adequate medical treatment.


Continuous treatment with cisplatin induces the oocyte death of primordial follicles without activation.
Chemotherapy directly or indirectly affects organs in a short-term or continuous manner. Endocrine organs are especially sensitive to cancer treatment, leading to concerns among patients regarding their quality of life afterward. Side effects to the ovary include damage to the ovarian reserve, resulting in follicle loss, endocrine hormone deficiency, and infertility. It has been previously demonstrated that continuous treatment with 2 mg/kg cisplatin for 15 days can activate primordial follicles, suggesting that the response in the oocytes of primordial follicles was dependent on cisplatin concentration and administration frequency. However, our results demonstrate that continuous treatment with 2 mg/kg cisplatin for 15 days leads to the same consequence as with the continuous treatment of 5 mg/kg cisplatin: the death of oocytes in primordial follicles without indication of activation. Moreover, animals co-injected with melatonin and cisplatin did not display any significant differences from those treated with cisplatin only contrary to the known results. 6-hydroxymelatonin, a metabolite of melatonin, could not prevent follicle destruction, implying that melatonin does not confer the protection of ovarian follicles, either directly or indirectly. Altogether, our data support that fertoprotectants against cisplatin must target molecules that control cell death pathways in the oocytes of primordial follicles.


Hypoglycaemia following upper gastrointestinal surgery: case report and review of the literature.
Hyperinsulinemic hypoglycemia is relatively recently recognized in persons undergoing bariatric surgery although knowledge and experience with this condition may not be commensurate with the number of such procedures being performed globally. This paper presents a novel case as an example of how such patients may present and how they may be investigated. A 69-year-old man was assessed 3 months post-fundoplication surgery for postprandial hypoglycaemia with neuroglycopenia that became progressively severe. A 72-h fast failed to show hypoglycaemia. During a clinic visit, the patient became confused and had a low plasma glucose, high plasma insulin, and high plasma C-peptide; symptoms were relieved with glucose. No tumours were visualized on CT, MRI, or endoscopic ultrasound. A total body Indium111-octreotide scan was negative. Selective arterial calcium stimulation showed a high insulin gradient in the splenic and superior mesenteric arteries, suggesting diffuse pancreatic beta cell hyperplasia. The patient declined pancreatic resection and recurrent symptomatic hypoglycaemia was successfully prevented with low dose octreotide. Although increasingly recognized following bariatric surgery, this is the first reported development of NIPHS (non-insulinoma pancreatogenous hypoglycemia syndrome) following fundoplication surgery, as well as the first documented use of octreotide in post-operative NIPHS. Medical management may be an alternative to surgery for patients with this rare condition.


CD20dim-positive T-cell large granular lymphocytic leukemia in a patient with concurrent hairy cell leukemia and plasma cell myeloma.
We report a CD20dim- positive T-cell large granular lymphocytic (T-LGL) leukemia in a patient with concurrent hairy cell leukemia and plasma cell myeloma. This patient was first diagnosed with T-LGL leukemia with dim CD20 expression, which by itself was a rare entity. He received no treatment for T-LGL leukemia. The patient later developed a hairy cell leukemia, which went into complete clinical remission after one cycle of 2-CdA. Five years later, he was diagnosed with a third malignancy, plasma cell myeloma. Complex cytogenetic aberrancies were present at the time when plasma cell myeloma was diagnosed. This is the first report, to the best of our knowledge, in the English literature with the aforementioned three distinct hematopoietic malignancies in one patient.


Microfluidic heart on a chip for higher throughput pharmacological studies.
We present the design of a higher throughput "heart on a chip" which utilizes a semi-automated fabrication technique to process sub millimeter sized thin film cantilevers of soft elastomers. Anisotropic cardiac microtissues which recapitulate the laminar architecture of the heart ventricle are engineered on these cantilevers. Deflection of these cantilevers, termed Muscular Thin Films (MTFs), during muscle contraction allows calculation of diastolic and systolic stresses generated by the engineered tissues. We also present the design of a reusable one channel fluidic microdevice completely built out of autoclavable materials which incorporates various features required for an optical cardiac contractility assay: metallic base which fits on a heating element for temperature control, transparent top for recording cantilever deformation and embedded electrodes for electrical field stimulation of the tissue. We employ the microdevice to test the positive inotropic effect of isoproterenol on cardiac contractility at dosages ranging from 1 nM to 100 μM. The higher throughput fluidic heart on a chip has applications in testing of cardiac tissues built from rare or expensive cell sources and for integration with other organ mimics. These advances will help alleviate translational barriers for commercial adoption of these technologies by improving the throughput and reproducibility of readout, standardization of the platform and scalability of manufacture.


Cold Inducible RNA Binding Protein Is Involved in Chronic Hypoxia Induced Neuron Apoptosis by Down-Regulating HIF-1α Expression and Regulated By microRNA-23a.
Background: Neuron apoptosis mediated by hypoxia inducible factor 1α (HIF-1α) in hippocampus is one of the most important factors accounting for the chronic hypobaric hypoxia induced cognitive impairment. As a neuroprotective molecule that is up-regulated in response to various environmental stress, CIRBP was reported to crosstalk with HIF-1α under cellular stress. However, its function under chronic hypobaric hypoxia remains unknown. Objective: In this study, we tried to identify the role of CIRBP in HIF-1α mediated neuron apoptosis under chronic hypobaric hypoxia and find a possible method to maintain its potential neuroprotective in long-term high altitude environmental exposure. Methods: We established a chronic hypobaric hypoxia rat model as well as a tissue culture model where SH-SY5Y cells were exposed to 1% hypoxia. Based on these models, we measured the expressions of HIF-1α and CIRBP under hypoxia exposure and examined the apoptosis of neurons by TUNEL immunofluorescence staining and western blot analysis of apoptosis related proteins. In addition, by establishing HIF-1α shRNA and pEGFP-CIRBP plasmid transfected cells, we confirmed the role of HIF-1α in chronic hypoxia induced neuron apoptosis and identified the influence of CIRBP over-expression upon HIF-1α and neuron apoptosis in the process of exposure. Furthermore, we measured the expression of the reported hypoxia related miRNAs in both models and the influence of miRNAs' over-expression/knock-down upon CIRBP in the process of HIF-1α mediated neuron apoptosis. Results: HIF-1α expression as well as neuron apoptosis was significantly elevated by chronic hypobaric hypoxia both in vivo and in vitro. CIRBP was induced in the early stage of exposure (3d/7d); however as the exposure was prolonged (21d), CIRBP level of the hypoxia group became significantly lower than that of control. In addition, HIF-1α knockdown significantly decreased neuron apoptosis under hypoxia, suggesting HIF-1α may be pro-apoptotic in the process of exposure. CIRBP over-expression significantly suppressed HIF-1α up-regulation in hypoxia and inhibited HIF-1α mediated neuron apoptosis. Interestingly, miR-23a was also induced by hypoxia exposure and showed the same changing tendency with CIRBP (increasing in 3d/7d, decreasing in 21d). In addition, over-expressing miR-23a up-regulated CIRBP, down-regulated HIF-1α and attenuated neuron apoptosis. Conclusion: Cold inducible RNA binding protein is involved in chronic hypoxia induced neuron apoptosis by down-regulating HIF-1α expression, and MiR-23a may be an important tool to maintain CIRBP level and function.


Correlation and prediction of gene expression level from amino acid and dipeptide composition of its protein.
A large number of papers have been published on analysis of microarray data with particular emphasis on normalization of data, detection of differentially expressed genes, clustering of genes and regulatory network. On other hand there are only few studies on relation between expression level and composition of nucleotide/protein sequence, using expression data. There is a need to understand why particular genes/proteins express more in particular conditions. In this study, we analyze 3468 genes of Saccharomyces cerevisiae obtained from Holstege et al., (1998) to understand the relationship between expression level and amino acid composition. We compute the correlation between expression of a gene and amino acid composition of its protein. It was observed that some residues (like Ala, Gly, Arg and Val) have significant positive correlation (r > 0.20) and some other residues (Like Asp, Leu, Asn and Ser) have negative correlation (r < -0.15) with the expression of genes. A significant negative correlation (r = -0.18) was also found between length and gene expression. These observations indicate the relationship between percent composition and gene expression level. Thus, attempts have been made to develop a Support Vector Machine (SVM) based method for predicting the expression level of genes from its protein sequence. In this method the SVM is trained with proteins whose gene expression data is known in a given condition. Then trained SVM is used to predict the gene expression of other proteins of the same organism in the same condition. A correlation coefficient r = 0.70 was obtained between predicted and experimentally determined expression of genes, which improves from r = 0.70 to 0.72 when dipeptide composition was used instead of residue composition. The method was evaluated using 5-fold cross validation test. We also demonstrate that amino acid composition information along with gene expression data can be used for improving the function classification of proteins. There is a correlation between gene expression and amino acid composition that can be used to predict the expression level of genes up to a certain extent. A web server based on the above strategy has been developed for calculating the correlation between amino acid composition and gene expression and prediction of expression level http://kiwi.postech.ac.kr/raghava/lgepred/. This server will allow users to study the evolution from expression data.


Spleen hypoplasia leads to abnormal stress hematopoiesis in mice with loss of Pbx homeoproteins in splenic mesenchyme.
The spleen plays critical roles in immunity and also provides a permissive microenvironment for hematopoiesis. Previous studies have reported that the TALE-class homeodomain transcription factor Pbx1 is essential in hematopoietic stem and progenitor cells (HSPCs) for stem cell maintenance and progenitor expansion. However, the role of Pbx1 in the hematopoietic niche has not been investigated. Here we explored the effects that genetic perturbation of the splenic mesenchymal niche has on hematopoiesis upon loss of members of the Pbx family of homeoproteins. Splenic mesenchyme-specific inactivation of Pbx1 (SKO) on a Pbx2- or Pbx3-deficient genetic background (DKO) resulted in abnormal development of the spleen, which is dysmorphic and severely hypoplastic. This phenotype, in turn, affected the number of HSPCs in the fetal and adult spleen at steady state, as well as markedly impairing the kinetics of hematopoietic regeneration in adult mice after sub-lethal and lethal myelosuppressive irradiation. Spleens of mice with compound Pyx deficiency 8 days following sublethal irradiation displayed significant downregulation of multiple cytokine-encoding genes, including KitL/SCF, Cxcl12/SDF-1, IL-3, IL-4, GM-CSF/Csf2 IL-10, and Igf-1, compared with controls. KitL/SCF and Cxcl12/SDF-1 were recently shown to play key roles in the splenic niche in response to various haematopoietic stresses such as myeloablation, blood loss, or pregnancy. Our results demonstrate that, in addition to their intrinsic roles in HSPCs, non-cell autonomous functions of Pbx factors within the splenic niche contribute to the regulation of hematopoiesis, at least in part via the control of KitL/SCF and Cxcl12/SDF-1. Furthermore, our study establishes that abnormal spleen development and hypoplasia have deleterious effects on the efficiency of hematopoietic recovery after bone marrow injury.


The metabolic consequences of altered glucose transporter expression in transgenic mice.
Glucose transporters are a family of membrane proteins which mediate glucose uptake across the cell membrane. The facilitative glucose transporter proteins are products of unique genes and are expressed in a tissue-specific manner. They are very similar structurally, containing 12 putative membrane spanning domains. Functionally they vary in their affinity for glucose and sensitivity to hormones such as insulin. Glucose homeostasis depends mainly on controlled changes in glucose transport in insulin-responsive tissues such as skeletal muscle and adipose cells where both glucose transporter 1 and glucose transporter 4 are expressed. Glucose transporter 4 is the major glucose transporter in these tissues and translocates from an intracellular vesicle to the cell membrane in response to insulin. Alterations of the level of expression of these glucose transporters should result in changes in insulin sensitivity and modification of whole-body metabolism. To test these hypotheses transgenic mouse models have been generated which overexpress glucose transporters in specific tissues or in the whole body. Glucose transporter 1 and glucose transporter 4 have been overexpressed specifically in skeletal muscle and glucose transporter 4 specifically in adipose tissue. Mice have also been made which overexpress glucose transporter 4 in the whole body. Using homologous recombination technology to disrupt the glucose transporter 4 gene, a "knockout" mouse has been created which expresses no glucose transporter 4. The metabolic consequences of these genetic manipulations on the level of expression of glucose transporters in the mouse are reviewed. The future applications of transgenic mouse technology in creating models which mimic human diseases are also discussed.


Optimizing rating scale category effectiveness.
Rating scales are employed as a means of extracting more information out of an item than would be obtained from a mere "yes/no", "right/wrong" or other dichotomy. But does this additional information increase measurement accuracy and precision? Eight guidelines are suggested to aid the analyst in optimizing the manner in which rating scales categories cooperate in order to improve the utility of the resultant measures. Though these guidelines are presented within the context of Rasch analysis, they reflect aspects of rating scale functioning which impact all methods of analysis. The guidelines feature rating-scale-based data such as category frequency, ordering, rating-to-measure inferential coherence, and the quality of the scale from measurement and statistical perspectives. The manner in which the guidelines prompt recategorization or reconceptualization of the rating scale is indicated. Utilization of the guidelines is illustrated through their application to two published data sets.


Self-associated molecular patterns mediate cancer immune evasion by engaging Siglecs on T cells.
First-generation immune checkpoint inhibitors, including anti-CTLA-4 and anti-programmed death 1 (anti-PD-1) antibodies, have led to major clinical progress, yet resistance frequently leads to treatment failure. Thus, new targets acting on T cells are needed. CD33-related sialic acid-binding immunoglobulin-like lectins (Siglecs) are pattern-recognition immune receptors binding to a range of sialoglycan ligands, which appear to function as self-associated molecular patterns (SAMPs) that suppress autoimmune responses. Siglecs are expressed at very low levels on normal T cells, and these receptors were not until recently considered as interesting targets on T cells for cancer immunotherapy. Here, we show an upregulation of Siglecs, including Siglec-9, on tumor-infiltrating T cells from non-small cell lung cancer (NSCLC), colorectal, and ovarian cancer patients. Siglec-9-expressing T cells coexpressed several inhibitory receptors, including PD-1. Targeting of the sialoglycan-SAMP/Siglec pathway in vitro and in vivo resulted in increased anticancer immunity. T cell expression of Siglec-9 in NSCLC patients correlated with reduced survival, and Siglec-9 polymorphisms showed association with the risk of developing lung and colorectal cancer. Our data identify the sialoglycan-SAMP/Siglec pathway as a potential target for improving T cell activation for immunotherapy.


Intrinsic functional connectivity in social anxiety disorder with and without comorbid attention deficit hyperactivity disorder.
Neuroimaging research about social anxiety disorder (SAD) points to hyperactivity in the fear circuit and altered connectivity between the fear circuit and the intrinsic connectivity networks that modulate it. We investigated intrinsic functional connectivity changes in SAD patients by taking into consideration the commonly overlooked comorbidity of attention deficit hyperactivity disorder (ADHD). We compared intrinsic functional connectivity alterations in 16 patients with pure SAD, 18 patients with SAD and comorbid ADHD and 21 healthy controls using seed-to-voxel functional connectivity analyses. Hypoconnectivity of the right fusiform gyrus with the left lingual gyrus was the unique difference between whole SAD group and healthy controls, while in the pure SAD group the fusiform gyrus displayed hypoconnectivity with the posterior default mode network (DMN) regions. In contrast, ADHD comorbidity was associated with hyperconnectivities of the salience network (SN) with the fusiform cortex and the posterior DMN regions, and hyperconnectivities of the posterior DMN with visual, somatosensory and motor cortices. The dichotomic dissociation of the SAD related functional connectivity changes into hypoconnectivities in the pure SAD group vs hyperconnectivities in the SAD-ADHD group leads also to the question, whether ADHD treatment can be considered an alternative for selected SAD cases.


A Bayesian Approach to Predict Food Fraud Type and Point of Adulteration.
Primary and secondary food processing had been identified as areas vulnerable to fraud. Besides the food processing area, other stages within the food supply chain are also vulnerable to fraud. This study aims to develop a Bayesian network (BN) model to predict food fraud type and point of adulteration i.e., the occurrence of fraudulent activity. The BN model was developed using GeNie Modeler (BayesFusion, LLC) based on 715 notifications (1979-2018) from Food Adulteration Incidents Registry (FAIR) database. Types of food fraud were linked to six explanatory variables such as food categories, year, adulterants (chemicals, ingredients, non-food, microbiological, physical, and others), reporting country, point of adulteration, and point of detection. The BN model was validated using 80 notifications from 2019 to determine the predictive accuracy of food fraud type and point of adulteration. Mislabelling (20.7%), artificial enhancement (17.2%), and substitution (16.4%) were the most commonly reported types of fraud. Beverages (21.4%), dairy (14.3%), and meat (14.0%) received the highest fraud notifications. Adulterants such as chemicals (21.7%) (e.g., formaldehyde, methanol, bleaching agent) and cheaper, expired or rotten ingredients (13.7%) were often used to adulterate food. Manufacturing (63.9%) was identified as the main point of adulteration followed by the retailer (13.4%) and distribution (9.9%).


The intermediate filament vimentin regulates chondrogenesis of adult human bone marrow-derived multipotent progenitor cells.
Cytoskeletal proteins play important regulatory roles in a variety of cellular processes, including proliferation, migration, and differentiation. However, whereas actin and tubulin have established roles regulating developmental chondrogenesis, there is no evidence supporting a function for the intermediate filament vimentin in embryonic cartilage formation. We hypothesized that vimentin may regulate the chondrogenic differentiation of adult multipotent progenitor cells (MPCs), such as those involved in cartilage formation during bone fracture repair. As our model of adult progenitor cell chondrogenesis, we employed high-density pellet cultures of human bone marrow-derived MPCs. siRNA-mediated knockdown of vimentin mRNA and protein triggered a reduction in the extent of MPC cartilage formation, as evidenced by depressed accumulation of mRNAs for the cartilage-specific marker genes aggrecan and collagen type II, as well as reduced levels of Alcian blue-stainable proteoglycan and collagen II protein in the extracellular matrix. Moreover, mRNA and protein levels for the chondro-regulatory transcription factors SOX5, SOX6, and SOX9 were diminished by vimentin knockdown. Depleted cellular vimentin also induced a drastic reduction in PKA phosphorylation levels but did not affect the phosphorylation of multiple other chondro-regulatory kinases and transcription factors, including ERK1/2, p38, Smad2, and Smad1/5/8. Importantly, siRNA-mediated knockdown of PKA C-alpha mRNA and protein mimicked the reduction in chondrogenesis caused by diminished cellular vimentin. Finally, overexpression of vimentin in MPCs significantly enhanced the activity of a transfected collagen II promoter-luciferase reporter gene. In conclusion, we describe a novel role for the intermediate filament vimentin as a positive regulator of adult human bone marrow-derived MPC chondrogenesis.


Unsupervised texture segmentation using multichannel decomposition and hidden Markov models.
In this paper, we describe an automatic unsupervised texture segmentation scheme using hidden Markov models (HMMs). First, the feature map of the image is formed using Laws' micromasks and directional macromasks. Each pixel in the feature map is represented by a sequence of 4-D feature vectors. The feature sequences belonging to the same texture are modeled as an HMM. Thus, if there are M different textures present in an image, there are M distinct HMMs to be found and trained. Consequently, the unsupervised texture segmentation problem becomes an HMM-based problem, where the appropriate number of HMMs, the associated model parameters, and the discrimination among the HMMs become the foci of our scheme. A two-stage segmentation procedure is used. First, coarse segmentation is used to obtain the approximate number of HMMs and their associated model parameters. Then, fine segmentation is used to accurately estimate the number of HMMs and the model parameters. In these two stages, the critical task of merging the similar HMMs is accomplished by comparing the discrimination information (DI) between the two HMMs against a threshold computed from the distribution of all DI's. A postprocessing stage of multiscale majority filtering is used to further enhance the segmented result. The proposed scheme is highly suitable for pipeline/parallel implementation. Detailed experimental results are reported. These results indicate that the present scheme compares favorably with respect to other successful schemes reported in the literature.


Establishment of the mammalian membrane block to polyspermy: evidence for calcium-dependent and -independent regulation.
One crucial result of egg activation is the establishment of blocks on the zona pellucida and the egg plasma membrane to prevent fertilization by additional sperm. The mechanism(s) by which a mammalian egg regulates the establishment of the membrane block to polyspermy is largely unknown. Since Ca(2+) signaling regulates several egg activation events, this study investigates how sperm-induced Ca(2+) transients affect the membrane block to polyspermy, building on our previous work (Biology of Reproduction 67:1342). We demonstrate that mouse eggs that experience only one sperm-induced Ca(2+) transient establish a membrane block that is less effective, than in eggs that experience normal sperm-induced Ca(2+) transients but that is more effective than in eggs with completely suppressed [Ca(2+)](cyt) increases. Sperm-induced increases in [Ca(2+)](cyt) regulate the timing of membrane block establishment, as this block is established more slowly in eggs that experience one or no sperm-induced Ca(2+) transients. Finally, our studies produce the intriguing discovery that there is also a Ca(2+)-independent event that is associated with fertilization in the pathway leading to membrane block establishment. Taken together, these data indicate that Ca(2+) plays a role in facilitating membrane block establishment by regulating the timing with which this change in egg membrane function occurs, and also that the membrane block differs from other post-fertilization egg activation responses as Ca(2+) is not the only stimulus. The membrane block to polyspermy in mammalian eggs is likely to be the culmination of multiple post-fertilization events that together modify the egg membrane's receptivity to sperm.


Deciphering SARS-CoV-2 Virologic and Immunologic Features.
Severe acute respiratory syndrome coronavirus (SARS-CoV)-2 and its associated pathology, COVID-19, have been of particular concerns these last months due to the worldwide burden they represent. The number of cases requiring intensive care being the critical point in this epidemic, a better understanding of the pathophysiology leading to these severe cases is urgently needed. Tissue lesions can be caused by the pathogen or can be driven by an overwhelmed immune response. Focusing on SARS-CoV-2, we and others have observed that this virus can trigger indeed an immune response that can be dysregulated in severe patients and leading to further injury to multiple organs. The purpose of the review is to bring to light the current knowledge about SARS-CoV-2 virologic and immunologic features. Thus, we address virus biology, life cycle, tropism for many organs and how ultimately it will affect several host biological and physiological functions, notably the immune response. Given that therapeutic avenues are now highly warranted, we also discuss the immunotherapies available to manage the infection and the clinical outcomes.


The reliability and predictive accuracy of the falls risk for older people in the community assessment (FROP-Com) tool.
the Falls Risk for Older People in the Community assessment (FROP-Com) tool was designed for use in targeted multi-factorial falls prevention programmes. It fills the gap between the short screening tools and the longer assessment tools, e.g. the physiological profile assessment (PPA). The aim of this study was to determine the reliability and predictive accuracy of the FROP-Com. the intra-rater and inter-rater reliability studies were performed with 20 participants each. The prospective study was performed with 344 community-dwelling older people presenting to an emergency department after a fall and being discharged directly home. Following a home-based assessment, including the FROP-Com, Timed Up and Go (TUG) and functional Reach (FR), participants were monitored for falls for 12 months. the intra-class correlation coefficient (ICC) for intra-rater reliability and inter-rater reliability for the FROP-Com was 0.93 (95% CI 0.84-0.97) and 0.81 (95% CI 0.59-0.92) respectively. The AUC for the FROP-Com was 0.68 (95% CI 0.63-0.74). At the cut-off 18/19, sensitivity was 71.3% (95% CI 64.4-78.3) and specificity was 56.1% (95% CI 48.9-63.4). The AUC for the TUG was 0.63 (95% CI 0.57-0.69) and for the FR was 0.60 (95% CI 0.54-0.66). the FROP-Com demonstrated good reliability and a moderate capacity to predict falls.


Improved designs for dose escalation studies using pharmacokinetic measurements.
We describe a method for incorporating pharmacokinetic (PK) data into dose escalation clinical trial designs. Doing so can improve the efficiency and accuracy of these studies. The method proposed uses a parametric dose response function that models the probability of response in each person with two effects: the dose of drug administered and an ancillary pharmacokinetic measurements. After treatment and observation of each subject (or group of subjects) for response, one calculates the dose to be administered to the next individual (or group) to yield the target probability of response from the current best estimate of the dose-response curve. This procedure is a variant of the continual reassessment method (CRM). Statistical simulations employing a logistic dose-response model (that is, we model the logit of the response probability as a linear combination of predictors), dose of drug, and the area under the time-concentration curve (AUC) demonstrate that the addition of pharmacokinetic information to the CRM is a practical and useful way to improve both dose-response modelling and the design of dose escalation studies.


Effects of the number of genome segments on primary and systemic infections with a multipartite plant RNA virus.
Multipartite plant viruses were discovered because of discrepancies between the observed dose response and predictions of the independent-action hypothesis (IAH) model. Theory suggests that the number of genome segments predicts the shape of the dose-response curve, but a rigorous test of this hypothesis has not been reported. Here, Alfalfa mosaic virus (AMV), a tripartite Alfamovirus, and transgenic Nicotianatabacum plants expressing no (wild type), one (P2), or two (P12) viral genome segments were used to test whether the number of genome segments necessary for infection predicts the dose response. The dose-response curve of wild-type plants was steep and congruent with the predicted kinetics of a multipartite virus, confirming previous results. Moreover, for P12 plants, the data support the IAH model, showing that the expression of virus genome segments by the host plant can modulate the infection kinetics of a tripartite virus to those of a monopartite virus. However, the different types of virus particles occurred at different frequencies, with a ratio of 116:45:1 (RNA1 to RNA2 to RNA3), which will affect infection kinetics and required analysis with a more comprehensive infection model. This analysis showed that each type of virus particle has a different probability of invading the host plant, at both the primary- and systemic-infection levels. While the number of genome segments affects the dose response, taking into consideration differences in the infection kinetics of the three types of AMV particles results in a better understanding of the infection process.


Adrenocortical and nociceptive responses to opioid blockade in hypertension-prone men and women.
Attenuated pain sensitivity and exaggerated adrenocortical stress reactivity have been documented in individuals at high risk for hypertension. The endogenous opioid system may play a role in these response alterations. We compared adrenocortical and nociceptive responses to opioid blockade using naltrexone in hypertension-prone men and women. Ninety-nine participants completed two sessions during which a placebo or 50 mg naltrexone was administered using a double-blind, counterbalanced design. Participants rated their pain and completed the McGill Pain Questionnaire (MPQ) after three assessments of the nociceptive flexion reflex and after assessment of nociceptive pain threshold and tolerance. Saliva samples were obtained throughout the sessions. Salivary cortisol levels increased after pain assessment after the ingestion of naltrexone, but not after placebo, with the low-risk group exhibiting an earlier peak of cortisol response. Participants reported greater pain ratings and higher MPQ scores in the naltrexone versus placebo condition, and these effects were more pronounced in women. Pain threshold and tolerance were higher among high-risk men relative to low-risk men. The results are consistent with the inhibitory effects of the endogenous opioids on cortisol response and suggest an altered response timeline among hypertension-prone individuals. The results demonstrate that hypoalgesia may be a marker of hypertension risk in men but not in women.


CRY2 suppresses trophoblast migration and invasion in recurrent spontaneous abortion.
Disruption of circadian rhythms is associated with aberrant trophoblast migration and invasion in recurrent spontaneous abortion (RSA). This study aims to explore the functional role and the mechanisms of cryptochrome 2 (CRY2), a fundamental component of the circadian clock, in regulating trophoblast migration and invasion. Human extravillous trophoblast cell line HTR-8/SVneo was used as a cell model. Cell migration and invasion were examined using wound healing assay and Transwell assay, respectively. The mRNA and protein levels were determined using quantitative real-time polymerase chain reaction and western blot, respectively. Luciferase reporter assay and chromatin immunoprecipitation assay were performed to explore the interaction between c-Myc to the brain and muscle ARNT-like protein 1 (BMAL1) promoter. CRY2 was highly expressed in human villous specimens of RSA. Furthermore, CRY2 overexpression impaired migration and invasion in HTR-8/SVneo cells, whereas CRY2 knockdown yielded the opposite results. Mechanistically, c-Myc bound to the BMAL1 promoter and induced BMAL1 transcription, both of which further activated matrix metalloproteinase 2/9 (MMP2/9) and facilitated migration and invasion in HTR-8/SVneo cells. CRY2 inhibited c-Myc-BMAL1 pathway and impaired migration and invasion of HTR-8/SVneo cells. Collectively, these findings demonstrate that CRY2 suppresses trophoblast migration and invasion via inhibiting c-Myc-BMAL1-MMP2/9 pathway.


Survival results of postoperative coronary angiogram for treatment of perioperative myocardial ischaemia following coronary artery bypass grafting: a single-centre experience.
Although perioperative myocardial ischaemia (PMI) is a well-known complication following coronary artery bypass grafting (CABG), standard strategies for its diagnosis and treatment are so far not defined. In this study, we sought to evaluate the impact on survival of postoperative coronary angiogram for management of patients with PMI after CABG. Overall, 4028 patients underwent isolated CABG in a single-centre institution between January 2006 and September 2013. A total of 168 (4.2%) patients received postoperative coronary angiogram because of diagnosis of PMI. These patients were matched on the basis of gender, age at surgery and date of surgery, with 336 (1:2 ratio) CABG patients without PMI to determine the impact of the PMI management. A total of 476 grafts were examined (263 venous grafts, 196 internal mammary artery grafts and 17 radial artery grafts). Almost three-quarters of the 168 PMI (74.4%) patients underwent postoperative coronary angiogram within 24 h of surgery. Normal postoperative coronary angiogram, graft failure and new native vessels occlusion were observed in 23.2%, 52.4% and 24.4% of patients, respectively. A total of 30 (17.9%) patients underwent surgical revision of grafts, whereas 60 (35.7%) patients were treated with percutaneous coronary intervention. Eighteen (10.7%) PMI patients died during the hospital stay compared with 6 (1.8%) patients in the non-PMI group. Survival rates at 7 years were 62.5% in the PMI group and 81.1% in non-PMI group (P < 0.001). After multivariable adjustment PMI (P < 0.001; hazard ratio 3.17, 95% confidence interval 2.12-4.73) turned out to be an independent predictor of mortality on follow-up. Moreover, further subanalysis revealed that delayed postoperative coronary angiogram (>24 h after surgery) was an independent predictor of poorer mid-term survival (P = 0.008; hazard ratio 3.62, 95% confidence interval 1.41-9.33). PMI after CABG is associated with a significantly poorer survival. A prompt postoperative management must always be considered. Further prospective studies are required to confirm our results.


Don't worry, be happy: Habitat selection of Blanding's Turtles (Emydoidea blandingii) living in a reference condition in Georgian Bay.
Few areas within the Great Lakes basin are currently free from impact of human activities, and it is important to study these reference conditions for comparison with degraded sites in those regions. Here, we use radio telemetry to investigate habitat use, movement, and habitat selection of a population of the endangered (Federally in Canada) Blanding's turtle (Emydoidea blandingii, BLTU) inhabiting a mostly undisturbed archipelago located at the northern shore of Mnidoo gamii (Georgian Bay), Ontario over two active seasons (May to September 2021 and 2022). We found a mean home range of 16.21 ha for females (n = 7) and 15.10 ha for males (n = 7). Of the five habitat classes (Marsh, Open Water, Rock, Peatland, and Forest), females used all except Peatland during the nesting season, and both sexes used all five habitat classes throughout both active seasons in 2021 and 2022. Disproportionate habitat use was detected at the landscape scale but not at the home range scale which was consistent with the hypothesis that adult Blanding's turtles residing in relatively undisturbed sites with abundant habitat types use all habitat types according to their availability. We also observed the use of open, deep water by Blanding's Turtles as travel corridors for nesting and mating. Effective future conservation strategies should prioritize the protection and connectivity of relatively undisturbed wetlands, forests, and rock barrens in this region and use this study as a reference condition to compare BLTU habitat use and movement across disturbance gradients within Georgian Bay.