Control of transient, resurgent, and persistent current by open-channel block by Na channel beta4 in cultured cerebellar granule neurons.
Voltage-gated Na channels in several classes of neurons, including cells of the cerebellum, are subject to an open-channel block and unblock by an endogenous protein. The Na(V)beta4 (Scn4b) subunit is a candidate blocking protein because a free peptide from its cytoplasmic tail, the beta4 peptide, can block open Na channels and induce resurgent current as channels unblock upon repolarization. In heterologous expression systems, however, Na(V)beta4 fails to produce resurgent current. We therefore tested the necessity of this subunit in generating resurgent current, as well as its influence on Na channel gating and action potential firing, by studying cultured cerebellar granule neurons treated with siRNA targeted against Scn4b. Knockdown of Scn4b, confirmed with quantitative RT-PCR, led to five electrophysiological phenotypes: a loss of resurgent current, a reduction of persistent current, a hyperpolarized half-inactivation voltage of transient current, a higher rheobase, and a decrease in repetitive firing. All disruptions of Na currents and firing were rescued by the beta4 peptide. The simplest interpretation is that Na(V)beta4 itself blocks Na channels of granule cells, making this subunit the first blocking protein that is responsible for resurgent current. The results also demonstrate that a known open-channel blocking peptide not only permits a rapid recovery from nonconducting states upon repolarization from positive voltages but also increases Na channel availability at negative potentials by antagonizing fast inactivation. Thus, Na(V)beta4 expression determines multiple aspects of Na channel gating, thereby regulating excitability in cultured cerebellar granule cells.


Two minimal Tat translocases in Bacillus.
Activity of the Tat machinery for protein transport across the inner membrane of Escherichia coli and the chloroplast thylakoidal membrane requires the presence of three membrane proteins: TatA, TatB and TatC. Here, we show that the Tat machinery of the Gram-positive bacterium Bacillus subtilis is very different because it contains at least two minimal Tat translocases, each composed of one specific TatA and one specific TatC component. A third, TatB-like component is apparently not required. This implies that TatA proteins of B. subtilis perform the functions of both TatA and TatB of E. coli and thylakoids. Notably, the two B. subtilis translocases named TatAdCd and TatAyCy both function as individual, substrate-specific translocases for the twin-arginine preproteins PhoD and YwbN, respectively. Importantly, these minimal TatAC translocases of B. subtilis are representative for the Tat machinery of the vast majority of Gram-positive bacteria, Streptomycetes being the only known exception with TatABC translocases.


Terahertz emission from ultrafast ionizing air in symmetry-broken laser fields.
A transient photocurrent model is developed to explain coherent terahertz emission from air irradiated by a symmetry-broken laser field composed of the fundamental and its second harmonic laser pulses. When the total laser field is asymmetric across individual optical cycles, a nonvanishing electron current surge can arise during optical field ionization of air, emitting a terahertz electromagnetic pulse. Terahertz power scalability is also investigated, and with optical pump energy of tens of millijoules per pulse, peak terahertz field strengths in excess of 150 kV/cm are routinely produced.


Freshmen weight and body composition change determinants: A scoping review.
Scoping review of freshmen weight and body composition change determinants in order to inform practice, policymaking, and research that improve health and well-being. Methods: A systematic search of the literature identified potential sources. Duplicates were removed before a title and abstract review. A full-text review was then conducted on the remaining sources. Retained sources were then reviewed systematically before synthesis. Results: Eighty-five sources were synthesized. Variation in study design, measurement, reporting, and analyses of determinants, in particular dietary characteristics and physical activity, complicate comparisons. Dietary characteristics and physical activity appear to influence freshmen weight and body composition changes, while evidence indicates alcohol consumption is associated with deleterious weight and body composition changes. Conclusions: Design, measurement, analyses, and reporting can be improved considerably to better examine relationship between body composition changes and determinants in order to provide insight into, and inform, interventions and policies to benefit students' health and well-being.


Combined transabdominal-transanal surgical approach for iatrogenic rectovaginal fistula: two case reports.
Rectovaginal fistula (RVF) is a type of anastomotic leakage that may occur after low anterior resection for rectal cancer. The repair of RVF can be challenging because of the scar tissue stenosis and incomplete obstruction. Two patients presented in our department with vaginal faecal discharge almost 7 months after the radical resection of rectal cancer. On vaginal examination, titanium nails related to the rectal surgery were found in the vaginal wall. The patients were diagnosed with RVF. Considering that RVF positions in the patients were high and might adhere to the pelvic tissue, a combined transabdominal-transanal resection and vaginal repair surgery was performed. About 3 months after surgery, both patients underwent colonic closure surgery, with consequent good recovery. A combined transabdominal-transanal approach may provide distinct advantages in surgical repair of difficult cases of RVF.


[Trematodes of birds (Aves) from the Middle Volga region. 2. Orders Plagiorchiida, Renicolida, Strigeida, and Schistosomatida].
Data on trematodes of Plagiorchiida, Renicolida, Strigeida, and Schistosomatida, parasitizing in birds of the Middle Volga region are given. Proceedings of the different authors are supplemented by results of our research. Two species of trematodes (Brachylaima mesostoma and Mosesia amplavaginata) are specified for birds of the middle Volga region for the first time. New hosts were revealed for 2 trematode species: the great tit for Plagiorchis laricola and the common chaffinch for M. amplavaginata. The following characteristics for each parasite are given: it's systematic position, the spectrum of hosts, localization, collecting site, biology, the degree of host specificity and geographical range. The morphological description and original figures of 11 species of trematodes are presented.


Data and sample sharing as an enabler for large-scale biomarker research and development: The EPND perspective.
Biomarker discovery, development, and validation are reliant on large-scale analyses of high-quality samples and data. Currently, significant quantities of data and samples have been generated by European studies on Alzheimer's disease (AD) and other neurodegenerative diseases (NDD), representing a valuable resource for developing biomarkers to support early detection of disease, treatment monitoring, and patient stratification. However, discovery of, access to, and sharing of data and samples from AD and NDD research are hindered both by silos that limit collaboration, and by the array of complex requirements for secure, legal, and ethical sharing. In this Perspective article, we examine key challenges currently hampering large-scale biomarker research, and outline how the European Platform for Neurodegenerative Diseases (EPND) plans to address them. The first such challenge is a fragmented landscape filled with technical barriers that make it difficult to discover and access high-quality samples and data in one location. A second challenge is related to the complex array of legal and ethical requirements that must be navigated by researchers when sharing data and samples, to ensure compliance with data protection regulations and research ethics. Another challenge is the lack of broad-scale collaboration and opportunities to facilitate partnerships between data and sample contributors and researchers, in addition to a lack of regulatory engagement early in the research process to enable validation of potential biomarkers. A further challenge facing projects is the need to remain sustainable beyond initial funding periods, ensuring data and samples are shared and reused, thereby driving further research and innovation. In addressing these challenges, EPND will enable an environment of faster and more disruptive research on diagnostics and disease-modifying therapies for Alzheimer's disease and other neurodegenerative diseases.


Rheumatic fever and smooth muscle antibodies.
Rheumatic fever is a nonsuppurative sequelae to Group A streptococcal pharyngitis. Valvulitis resulting from cardiac involvement in this disease, is the main cause of morbidity in rheumatic fever. The study of rheumatic fever sera on cryostat sections of a variety of tissues using indirect immunofluorescence is described. A 95% incidence of smooth muscle antibody was found in 236 rheumatic fever sera, from 55 patients. Sera from hospital patients with a variety of diseases had an overall incidence of 12% for smooth muscle antibodies. The pattern and distribution of tissue staining strongly suggests that the smooth muscle antibodies described in rheumatic fever sera have antiactin specificity. This finding raises many questions concerning the immunopathogenesis and immunogenetics of rheumatic fever, and the pathogenesis of a number of diseases in which streptococci have not previously been considered.


HLA-A*0201 restricted CD8+ T-lymphocyte responses to malaria: identification of new Plasmodium falciparum epitopes by IFN-gamma ELISPOT.
The role of antigen specific CD8+ T-lymphocytes in mediating protection against sporozoite-induced malaria has been well established in murine models. In humans, indirect evidence has accumulated suggesting a similar protective role for antigen-specific CD8+ T-lymphocytes. Nevertheless, the low frequency of circulating specific cells together with the lack of sensitive methods to quantify them has hampered the direct assessment of their function. Using a combination of short-term cell culture and IFN-gamma ELISPOT, we studied CD8+ T-lymphocyte responses to a panel of HLA-A*0201 binding peptides. In addition to confirming the response to already described epitopes, we also identified five new CD8+ T-lymphocyte epitopes. These epitopes are presented in pre-erythrocytic stages gene products of Plasmodium falciparum 7G8 strain and correspond to the following protein segments: circumsporozoite (CS) 64-72, 104-113, 299-308 and 403-411; liver stage antigen (LSA-1) repeat region; sporozoite surface protein 2 or thrombospondin related anonymous protein (SSP2/TRAP) 78-88 and 504-513. Four of these peptides are conserved amongst all published sequences of P. falciparum strains. We conclude that the modified IFN-gamma ELISPOT assay is a sensitive technique to monitor antigen-specific CD8+ T-lymphocyte responses in human malaria which may help in the improvement and assessment of the efficacy of malaria subunit vaccines.


Human Cytomegalovirus Inhibits the PARsylation Activity of Tankyrase--A Potential Strategy for Suppression of the Wnt Pathway.
Human cytomegalovirus (HCMV) was reported to downregulate the Wnt/β-catenin pathway. Induction of Axin1, the negative regulator of the Wnt pathway, has been reported as an important mechanism for inhibition of β-catenin. Since Tankyrase (TNKS) negatively regulates Axin1, we investigated the effect of HCMV on TNKS expression and poly-ADP ribose polymerase (PARsylation) activity, during virus replication. Starting at 24 h post infection, HCMV stabilized the expression of TNKS and reduced its PARsylation activity, resulting in accumulation of Axin1 and reduction in its PARsylation as well. General PARsylation was not changed in HCMV-infected cells, suggesting specific inhibition of TNKS PARsylation. Similarly, treatment with XAV939, a chemical inhibitor of TNKS' activity, resulted in the accumulation of TNKS in both non-infected and HCMV-infected cell lines. Reduction of TNKS activity or knockdown of TNKS was beneficial for HCMV, evidenced by its improved growth in fibroblasts. Our results suggest that HCMV modulates the activity of TNKS to induce Axin1, resulting in inhibition of the β-catenin pathway. Since HCMV replication is facilitated by TNKS knockdown or inhibition of its activity, TNKS may serve as an important virus target for control of a variety of cellular processes.


First description of Vibrio harveyi as the causative agent of morbidity and mortality in farmed flathead grey mullet Mugil cephalus.
Flathead grey mullet Mugil cephalus is an important species in the aquaculture industry in the Mediterranean basin and throughout the world. During the last 10 yr, M. cephalus breeding stocks, larvae, and juveniles cultured in Eilat (Israel) have shown neurological signs such as uncoordinated circular swimming, while also presenting oral hemorrhages. Death follows days after the onset of the clinical signs, and mortality rates may reach 80% in some cases, causing high economical losses. Bacteriology isolations from different organs, including the brain, and a Koch's postulate experiment, confirmed Vibrio harveyi as the causative agent. Histological analyses showed the presence of the bacterium in different organs. However, in the brain, the bacterium was observed only within blood vessels and meninges. In some samples, mild to severe brain tissue damage was seen. In order to understand the virulence and lethality of V. harveyi, a median lethal dose was calculated, and the result was 106 colony-forming units fish-1. To the best of our knowledge, this is the first report that describes V. harveyi isolated from the brain of M. cephalus and validates it as an etiological agent causing neurological signs in this fish species.


Cloning of the mspA gene encoding a porin from Mycobacterium smegmatis.
Porins form channels in the mycolic acid layer of mycobacteria and thereby control access of hydrophilic molecules to the cell. We purified a 100 kDa protein from Mycobacterium smegmatis and demonstrated its channel-forming activity by reconstitution in planar lipid bilayers. The mspA gene encodes a mature protein of 184 amino acids and an N-terminal signal sequence. MALDI mass spectrometry of the purified porin revealed a mass of 19 406 Da, in agreement with the predicted mass of mature MspA. Dissociation of the porin by boiling in 80% dimethyl sulphoxide yielded the MspA monomer, which did not form channels any more. Escherichia coli cells expressing the mspA gene produced the MspA monomer and a 100 kDa protein, which had the same channel-forming activity as whole-cell extracts of M. smegmatis with organic solvents. These proteins were specifically detected by a polyclonal antiserum that was raised to purified MspA of M. smegmatis. These results demonstrate that the mspA gene encodes a protein of M. smegmatis, which assembles to an extremely stable oligomer with high channel-forming activity. Database searches did not reveal significant similarities to any other known protein. Southern blots showed that the chromosomes of fast-growing mycobacterial species contain homologous sequences to mspA, whereas no hybridization could be detected with DNA from slow growing mycobacteria. These results suggest that MspA is the prototype of a new class of channel-forming proteins.


Influence of rhenium speciation on the stability and activity of Re/Pd bimetal catalysts used for perchlorate reduction.
Recent work demonstrates reduction of aqueous perchlorate by hydrogen at ambient temperatures and pressures using a novel rhenium-palladium bimetal catalyst immobilized on activated carbon (Re/Pd-AC). This study examines the influence of Re speciation on catalyst activity and stability. Rates of perchlorate reduction are linearly dependent on Re content from 0-6 wt %, but no further increases are observed at higher Re contents. Surface-immobilized Re shows varying stability and speciation both in oxic versus H(2)-reducing environments and as a function of Re content. In oxic solutions, Re immobilization is dictated by sorption of the Re(VII) precursor, perrhenate (ReO(4)(-)), to activated carbon via electrostatic interactions. Under H(2)-reducing conditions, Re immobilization is significantly improved and leaching is minimized by ReO(4)(-) reduction to more reduced species on the catalyst surface. X-ray photoelectron spectroscopy shows two different Re binding energy states under H(2)-reducing conditions that correspond most closely to Re(V)/Re(IV) and Re(I) reference standards, respectively. The distribution of the two redox states varies with Re content, with the latter predominating at lower Re contents where catalyst activity is more strongly dependent on Re content. Results demonstrate that both lower Re contents and the maintenance of H(2)-reducing conditions are key elements in stabilizing the active Re surface species that are needed for sustained catalytic perchlorate treatment.


Recent estimates of the out-of-pocket expenditure on health care in Australia.
Objective The aims of this study were to estimate the average annual out-of-pocket (OOP) expenditure on health care by households in Australia in 2015-16, and to compare this with the estimate for 2009-10. Methods Data from the most recent Household Expenditure Survey (HES) conducted by the Australian Bureau of Statistics were used. Various statistical methods were used to estimate the annual OOP expenditures at the household and national levels. Results The average annual OOP expenditure was A$4290 per household, representing 5.8% of the amount spent on all goods and services. Private health insurance (PHI) premiums, although not a direct expenditure on health care, were 40.6% of the total OOP expenses. Of the remaining 59.4%, nearly half was spent on doctors and other health professionals, and approximately one-third was spent on medicines. Dental treatments and specialist consultations were the most expensive, whereas visits to general practitioners incurred the least OOP expenditure. Households with PHI (58.6%) spent fourfold more on health care than those not insured. Compared with the 2009-10 survey, the biggest increases were in the cost of PHI (50.7%) and copayments to specialists (34.8%) and other health professionals (42.0%). Conclusions OOP expenditure on health care as a proportion of the total household expenditure on all goods and services has increased by more than 25% between 2009-10 and 2015-16. What is known about the topic? Australian households incur OOP expenses for health care in Australia for a wide range of goods and services, such as copayments to doctors and other health professionals beyond the Medicare rebates, the cost of medicines and other pharmaceutical goods not covered entirely by the Pharmaceutical Benefits Scheme and PHI premiums. Although other estimates of OOP expenditure are available in official reports of the Australian Institute of Health and Welfare, they are based on administrative records rather than consumer reports, and cannot be disaggregated by item or the characteristics of households. What does this paper add? This paper provides detailed information on OOP expenditure on health care as reported by a probability sample of households interviewed for the HES conducted by the ABS during 2015-16. These estimates of OOP expenditure, based on consumer reports, add a further dimension to the information available from administrative records only. What are the implications for practitioners? Practitioners should take account of the effect of increasing copayments for their services, especially on patients belonging to the lower socioeconomic categories. Increasing copayments may lead to people foregoing medical care. Health planners and politicians should note the steady upward drift in OOP expenses and factor these into their policies for future funding of health care.


Duplication 6q24 leads to 6qter in an infant from a balanced paternal translocation.
Duplication of 6q24 leads to 6qter was identified by GTG banding in an infant girl whose father was a balanced translocation carrier 46,XY,t(3;6)(p26 leads to q2402). At birth and at 4 mo she had proportionate short stature, microcephaly, asymmetric micrognathia, bow-shaped upper vermilion, long upper lip, submucous cleft palate, antimongoloid slant of palpebral fissures, telecanthus, prominent eyes, short neck with anterior and lateral webbing, short sternum, overlapping toes, wrist contractures, and hypertonicity. Later she was noted to have psychomotor retardation. Eleven previously published cases and our patient suggest that duplication of 6q (involving at least 6q25 leads to 6qter) produces a highly characteristic syndrome.


Case report: differential pressure impressions for complete dentures.
A method of producing a differential pressure, secondary impression of a lower arch with a sharp bony ridge is described. The use of this technique allows the clinician to dictate where the load is to be borne by the mucosa.


Ten years of knowledge representation for health care (2009-2018): Topics, trends, and challenges.
In the last ten years, the international workshop on knowledge representation for health care (KR4HC) has hosted outstanding contributions of the artificial intelligence in medicine community pertaining to the formalization and representation of medical knowledge for supporting clinical care. Contributions regarding modeling languages, technologies and methodologies to produce these models, their incorporation into medical decision support systems, and practical applications in concrete medical settings have been the main contributions and the basis to define the evolution of this field across Europe and worldwide. Carry out a review of the papers accepted in KR4HC in the 2009-2018 decade, analyze and characterize the topics and trends within this field, and identify challenges for the evolution of the area in the near future. We reviewed the title, the abstract, and the keywords of the 112 papers that were accepted to the workshop, identified the medical and technological topics involved in these works, provided a classification of these papers in medical and technological perspectives and obtained the timeline of these topics in order to determine interest growths and declines. The experience of the authors in the field and the evidences after the review were the basis to propose a list of challenges of knowledge representation in health care for the future. The most generic knowledge representation methods are ontologies (31%), semantic web related formalisms (26%), decision tables and rules (19%), logic (14%), and probabilistic models (10%). From a medical informatics perspective, knowledge is mainly represented as computer interpretable clinical guidelines (43%), medical domain ontologies (26%), and electronic health care records (22%). Within the knowledge lifecycle, contributions are found in knowledge generation (38%), knowledge specification (24%), exception detection and management (12%), knowledge enactment (8%), temporal knowledge and reasoning (7%), and knowledge sharing and maintenance (7%). The clinical emphasis of knowledge is mainly related to clinical treatments (27%), diagnosis (13%), clinical quality indicators (13%), and guideline integration for multimorbid patients (12%). According to the level of development of the works presented, we distinguished four maturity levels: formal (22%), implementation (52%), testing (13%), and deployment (2%) levels. Some papers described technologies for specific clinical issues or diseases, mainly cancer (22%) and diseases of the circulatory system (20%). Chronicity and comorbidity were present in 10% and 8% of the papers, respectively. KR4HC is a stable community, still active after ten years. A persistent focus has been knowledge representation, with an emphasis on semantic-web ontologies and on clinical-guideline based decision-support. Among others, two topics receive growing attention: integration of computer-interpretable guideline knowledge for the management of multimorbidity patients, and patient empowerment and patient-centric care.


Tracing the fate of wastewater viruses reveals catchment-scale virome diversity and connectivity.
The discharge of wastewater-derived viruses in aquatic environments impacts catchment-scale virome composition. To explore this, we used viromic analysis of RNA and DNA virus-like particles to holistically track virus communities entering and leaving wastewater treatment plants and the connecting river catchment system and estuary. We reconstructed >40 000 partial viral genomes into 10 149 species-level groups, dominated by dsDNA and (+)ssRNA bacteriophages (Caudoviricetes and Leviviricetes) and a small number of genomes that could pose a risk to human health. We found substantial viral diversity and geographically distinct virus communities associated with different wastewater treatment plants. River and estuarine water bodies harboured more diverse viral communities in downstream locations, influenced by tidal movement and proximity to wastewater treatment plants. Shellfish and beach sand were enriched in viral communities when compared with the surrounding water, acting as entrapment matrices for virus particles. Extensive phylogenetic analyses of environmental-derived and reference sequences showed the presence of human-associated sapovirus GII in all sample types, multiple rotavirus A strains in wastewater and a diverse set of picorna-like viruses associated with shellfish. We conclude that wastewater-derived viral genetic material is commonly deposited in the environment and can be traced throughout the freshwater-marine continuum of the river catchment, where it is influenced by local geography, weather events and tidal effects. Our data illustrate the utility of viromic analyses for wastewater- and environment-based ecology and epidemiology, and we present a conceptual model for the circulation of all types of viruses in a freshwater catchment.


Activation of intestinal intraepithelial T lymphocytes in calves infected with Cryptosporidium parvum.
The objective of this study was to identify disease-related changes in lymphocyte populations within ileal mucosae of calves with cryptosporidiosis. Groups of five neonatal calves were orally infected at 3 days of age with 10(8) oocysts and maintained in enteric-pathogen-free conditions until clinical disease was established or until the animals had recovered from disease. Age-matched uninfected calves were used for comparison. Ileal mucosal lymphocytes were collected, quantitated, and phenotyped to determine whether changes in lymphocyte composition occurred in infected animals. We observed significantly larger numbers of intraepithelial CD8+ T lymphocytes in ileal mucosae from acutely infected calves compared with those from control animals. In addition, a proportion of intraepithelial CD4+ T cells from acutely infected calves coexpressed CD25, whereas there was an absence of coexpressed CD25 on CD4+ T cells from control calves. Ex vivo reverse transcriptase PCR of RNA from intraepithelial lymphocytes from control calves showed a cytokine expression pattern consisting of tumor necrosis factor alpha (TNF-alpha) and gamma interferon (IFN-gamma), while intraepithelial lymphocytes from calves with cryptosporidiosis expressed IFN-gamma but not TNF-alpha. Together, the results indicate that changes occur in the ileal intraepithelial lymphocyte population coincidently with Cryptosporidium parvum-induced enteric disease.


Graphitic and oxidised high pressure high temperature (HPHT) nanodiamonds induce differential biological responses in breast cancer cell lines.
Nanodiamonds have demonstrated potential as powerful sensors in biomedicine, however, their translation into routine use requires a comprehensive understanding of their effect on the biological system being interrogated. Under normal fabrication processes, nanodiamonds are produced with a graphitic carbon shell, but are often oxidized in order to modify their surface chemistry for targeting to specific cellular compartments. Here, we assessed the biological impact of this purification process, considering cellular proliferation, uptake, and oxidative stress for graphitic and oxidized nanodiamond surfaces. We show for the first time that oxidized nanodiamonds possess improved biocompatibility compared to graphitic nanodiamonds in breast cancer cell lines, with graphitic nanodiamonds inducing higher levels of oxidative stress despite lower uptake.


Headaches in the accident and emergency department.
Serious illness is not uncommon in patients attending accident and emergency departments with headache. Careful clinical assessment, focusing on characteristics of the headache, associated symptoms and certain examination findings, helps to identify those at risk, and permits the efficient management of everybody else.


Importance of tumor location and histology in familial risk of upper gastrointestinal cancers: a nationwide cohort study.
Familial clustering of upper gastrointestinal (UGI) cancers and the significance of family history has been addressed previously. We aimed to elucidate the familial risk based on the specified tumor location and histology. In the Swedish Family-Cancer Database, we determined the familial risk of UGI cancer patients diagnosed (1958-2015) with esophageal and gastric cancer by tumor location using standardized incidence ratios (SIRs). Risk of esophageal cancer in first-degree relatives (FDRs) of patients with esophageal cancer increased 2.4-fold (SIR 95% CI 2.0-2.8), whereas risk of esophageal cancer in cases with family history of cancer in the middle third of the esophagus increased 3.4-fold (SIR 95% CI 2.1-5.1). Risk of gastric cancer in FDRs increased 1.6-fold (SIR 95% CI 1.5-1.7), occurrence of concordant subsite gastric cancer in the antrum, body, and cardia was 5.5-fold (SIR 95% CI 2.4-11), 4.6-fold (SIR 95% CI 2.6-7.4), and 1.7-fold (SIR 95% CI 1.1-2.5), respectively. Familial risk of concordant histological subtype in esophageal cancer was 4.1-fold for squamous cell carcinoma (SIR 95% CI 3.2-5.2) and 3.6-fold for adenocarcinoma (SIR 95% CI 2.5-5.1). The risk of concordant gastric adenocarcinoma was 1.6-fold for one affected FDR (SIR 95% CI 1.5-1.7), 6.1-fold for two FDRs (SIR 95% CI 4.4-8.4), and 8.6-fold among twins (SIR 95% CI 2.3-22). Family history of cancer in the lower third of the esophagus and stomach cancer in specific locations such as the antrum, body, and cardia can be considered as important predictive evidence for cancer in the same location in relatives. Our findings might guide endoscopy-based surveillance by introducing subgroups of populations with a higher risk for UGI cancer with particular attention to concordance of location of lesions, which could be a reasonable strategy for early detection, and thus help save more lives.


The initiation of pre-ecdysis and ecdysis behaviors in larval Manduca sexta: the roles of the brain, terminal ganglion and eclosion hormone.
Each larval molt of Manduca sexta culminates in the sequential performance of pre-ecdysis (cuticle loosening) and ecdysis (cuticle shedding) behaviors. Both behaviors are thought to be triggered by the release of a peptide, eclosion hormone (EH), from brain neurons whose axons extend the length of the nervous system. EH bioactivity appears in the hemolymph at the onset of pre-ecdysis behavior, and EH injection can trigger pre-ecdysis and ecdysis behaviors prematurely. The present study examined the effects of removing or disconnecting portions of the central nervous system prior to the time of EH release on the initiation of pre-ecdysis and ecdysis behaviors at the final larval molt. We found that the initiation of pre-ecdysis abdominal compressions at the appropriate time required the terminal abdominal ganglion (AT) but not the brain; the initiation of pre-ecdysis proleg retractions at the appropriate time required neither the AT nor the brain; the initiation of ecdysis at the appropriate time usually required the brain but did not require the AT; and premature pre-ecdysis (but not ecdysis) could be elicited in isolated abdomens by injection of EH. Finally, pre-ecdysis behavior performed by brainless larvae was not associated with the normal elevation of EH bioactivity in the hemolymph or the normal loss of EH immunoreactivity from peripheral neurohemal release sites.


Effect on nutritional markers of a model of aberrant crypt foci induced by azoxymethane and sodium dextran sulfate in Sprague Dawley rats.
Introduction: aberrant crypt foci (ACF) are colon preneoplastic lesions that can be used as a tool to study preventive processes for colorectal cancer (CRC). This model consists of initiation induced by azoxymethane (AOM) and promoted by sodium dextran sulfate (DSS), simulating human colonic carcinogenesis in a rat model. There is no direct information on the effects of this process on nutritional markers. Objective: to determine the effect on nutritional markers after the induction of ACF by AOM/DSS in a rat model. Methods: ACF were induced in 24 four-week-old Sprague Dawley male rats by administration of 2 AOM injections (10 mg/kg) and 7 days of 2% DSS in their drinking water. Body weight gain, food and fluid intake, weight of sacrificial organs, nutritional biochemical profiles, liver and kidney toxicity were evaluated. Cell counts in blood were also performed and histological sections evaluated in specific organs. The model was confirmed with identification and counts of ACF. Half of the rats were sacrificed at the sub-chronic stage and the rest at the chronic stage. Results: at the sub-chronic stage, changes in the liver and colon weight, and in the lymphocyte count were observed. For both stages, histopathological damage was observed in liver, kidney and colon, along with alterations in serum glucose levels. Conclusions: the model for proposed ACF can be used at the sub-chronic stage without the need for observation at the chronic stage. More research is needed to determine the mechanism of the observed effects.


An uncontrolled trial of a present-focused cognitive-behavioral conjoint therapy for posttraumatic stress disorder.
The efficacy of a present-focused version of cognitive-behavioral conjoint therapy for posttraumatic stress disorder (CBCT for PTSD) was examined in a community sample. Seven couples completed pretreatment assessments, including measures of clinician-, self- and partner-rated PTSD symptoms and relationship satisfaction. Six couples completed present-focused CBCT for PTSD and all posttreatment assessments. A seventh couple terminated their relationship prior to completing treatment; therefore, they completed posttreatment symptom measures, but not ratings of relationship satisfaction. Results revealed significant decreases in PTSD symptoms that were associated with medium-to-large effect sizes. Medium effect sizes for changes in relationship satisfaction were found, though were only significant for partners. Results from this pilot study suggest that present-focused CBCT for PTSD may be a promising alternative for individuals who are unwilling to engage in a trauma-focused treatment.


Differential effect of GalNAcalpha-O-bn on intracellular trafficking in enterocytic HT-29 and Caco-2 cells: correlation with the glycosyltransferase expression pattern.
Our previous work has shown that long-term treatment of mucus-secreting HT-29 cells with 1-benzyl-2-acetamido-2-deoxy-alpha-D-galactopyranoside (GalNAcalpha-O-bn), a competitive inhibitor of O-glycosylation, induced several phenotypic changes, in particular a blockade in the secretion of mucins, which are extensively O-glycosylated glycoproteins. Here, we have analyzed the effects of GalNAcalpha-O-bn upon the intracellular trafficking of basolateral and apical membrane glycoproteins at the cellular and biochemical levels in two types of cells, HT-29 G(-) and Caco-2, differentiated into an enterocyte-like phenotype. In HT-29 G(-) cells, but not in Caco-2 cells, DPP-IV and CD44 failed to be targeted to the apical or basolateral membrane, respectively, and accumulated inside intracytoplasmic vesicles together with GalNAcalpha-O-bn metabolites. We observed a strong inhibition of alpha2,3-sialylation of glycoproteins in HT-29 G(-) cells correlated to the high expression of alpha2,3-sialyltransferases ST3Gal I and ST3Gal IV. In these cells, DPP-IV and CD44 lost the sialic acid residue substituting the O-linked core 1 structure Galbeta1-3GalNAc (T-antigen). In contrast, sialylation was not modified in Caco-2 cells, but a decrease of alpha1,2-fucosylation was observed, in correlation with the high expression of alpha1,2-fucosyltransferases Fuc-TI and Fuc-TII. In conclusion, in HT-29 G(-) cells, GalNAcalpha-O-bn induces a specific cellular phenotype, which is morphologically characterized by the formation of numerous intracellular vesicles, in which are accumulated defectively sialylated O-glycosylproteins originally targeted to basolateral or apical membranes, and GalNAcalpha-O-bn metabolites.


Focal Hepatic Glycogenosis in a Patient With Uncontrolled Diabetes Mellitus Type 1.
Hepatomegaly and elevated liver enzymes in patients with diabetes are commonly associated with fatty liver disease. However, physicians often forget about another intrinsic substance that can cause a similar clinical picture-glycogen. Liver stores approximately one third of the total body glycogen and is responsible for blood glucose homeostasis. Excessive hepatocellular glycogen accumulation occurs not only in congenital glycogen storage diseases, but also in acquired conditions associated with hyperglycemic-hyperinsulinemic states such as uncontrolled diabetes mellitus, high-dose corticosteroid use, and dumping syndrome. All reported cases of acquired abnormal glycogen deposition described a diffuse form of hepatic glycogenosis with the entire liver involved in the accumulating process. To our knowledge, this is the first reported case of abnormal focal glycogen deposition in a patient with diabetes mellitus type 1 with imaging and pathologic correlation. Awareness of the imaging appearance of focal glycogen deposition can help to distinguish it from other pathologic conditions.


Odorant receptor gene expression changes during the parr-smolt transformation in Atlantic salmon.
The ability of salmon to home accurately to their natal stream to spawn has long intrigued biologists and has important consequences for the maintenance of population structure in these species. It is known that olfaction is crucial to homing, and that the transition from the freshwater to the marine environment (the parr-smolt transformation; PST) is a period of increased olfactory sensitivity and learning, resulting in a permanent memory of natal site odours that is retained, at least in part, in peripheral sensory neurones. These odours are then used as cues by sexually maturing fish on their homeward migration. We used quantitative polymerase chain reaction techniques to demonstrate transient increases in expression of odorant receptor transcripts (of up to fifty-fold over pre-PST levels) coincident with PST. Both olfactory (SORB) and vomeronasal receptors (SVRA and SVRC) are involved, which suggests that the fish learn both environmental odours and semiochemicals (pheromones). Receptor expression varies between families and changes over time indicating both genetic differences in odour stimuli and multiple periods of olfactory sensitivity. We suggest that changes in OR gene expression may have a role in homing behaviour and thus the maintenance of population structure in Atlantic salmon.


[Luminescence laparoscopy in abdominal surgery].
Luminescent laparoscopy was performed in 51 cases for the assessment of the intestinal blood circulation. Sixteen examinations were performed in acute cholecystitis. Pseudo-negative result has been obtained in 4.5% of the 67 observations. The traditional technique of laparoscopy used with the same purpose in 113 examinations gave errors in 48.7%.


Life events and depression in women: a structural equation model.
A non-recursive structural equation model designed to examine the reciprocal relationship between life event reports and depressive symptoms in women is presented. The model was fitted to data from a 2-year longitudinal study of life events and depression in a sample of women with 5-year-old children. The findings suggest that life event reports may be influenced by mental state; however, even when this tendency is taken into account, there is still a substantial relationship between life events and depressive symptoms in women. This indicates that it is unlikely that the life events/depression correlation can be explained simply by a tendency for depressed women to over-report life events.


The relationship of lumbar multifidus muscle morphology to previous, current, and future low back pain: a 9-year population-based prospective cohort study.
Population based prospective cohort study. We explored the cross-sectional relationships between lumbar multifidus (LM) intramuscular adipose tissue (IMAT) infiltration and low back pain (LBP) at 3 successive time points and investigated the role of IMAT in predicting the occurrence of LBP after 5 and 9 years. Although LBP is a major source of disease burden, the biological determinants of LBP are poorly understood. Participants were 40-year-old adults randomly sampled from a Danish population and followed up at 45 and 49 years of age. At each time point, participants underwent magnetic resonance imaging and reported ever having had LBP, LBP in the previous year, nontrivial LBP in the previous year, or a history of pain radiating into the legs. Pixel intensity and frequencies from T1-weighted magnetic resonance images identified the greatest proportion of LM IMAT at the L4 and L5 spinal levels. IMAT infiltration was categorized as normal/mild, moderate, or severe based on tertile divisions. Associations were explored with crude and adjusted odds ratios (aORs) from logistic regression models. Model covariates included sex, body mass index, and occupational and leisure time physical activity. A total of 401 participants were enrolled, with 331 (83%) and 286 (71%) participants followed up at 5 and 9 years, respectively. The cross-sectional analyses demonstrated that at the age of 40 years, participants with severe IMAT infiltration demonstrated increased odds of ever experiencing LBP (aOR [95% confidence interval, 95% CI] = 3.16 [1.45-6.89]), nontrivial LBP (aOR [95% CI] = 2.82 [1.36-5.81]), LBP in the past year (aOR [95% CI] = 1.95 [1.07-3.53]), and leg pain (aOR [95% CI] = 2.08 [1.19-3.62]). There were no consistent cross-sectional associations between LBP/leg pain and LM IMAT at 45 or 49 years of age and LM IMAT did not predict future LBP or leg pain. The relationship between LM IMAT and LBP/leg pain is inconsistent and may be modified by age. N/A.


[Characterization of B1-cells during experimental leukomogenesis.]
Bovine leukemia causes a significant polyclonal expansion of CD5+, IgM+ B lymphocytes, known as persistent lymphocytosis (PL), in approximately 30% of infected cattle. However, it is not yet clear what happens to this subpopulation of B cells in the early period of infection of animals. Quantitative characterization of IgM+ and CD5+ B cells during the immune response, which can provide important information on the mechanisms of lymphocyte priming in BLV infection. The experiment used BLV-negative calves of black-motley breed at the age of 8 months (n = 11). Animals (n = 8) were intravenously injected with blood of a BLV-positive cow. Control calves (n = 3) were injected with saline. Studies were performed before and after infection on days 5, 7, 14, 21, 28 and 65 of the immune response. The determination of the number of B-lymphocytes in the blood was carried out by the method of immunoperoxidase staining based on monoclonal antibodies to IgM, CD5. As a result of the studies, it was found that the level of CD5+ B cells increases on the 14th day of the primary immune response, characterized by polyclonal proliferation of CD5+ B cells, which are the primary target for BLV. Our research data confirm that in the lymphocytes of experimentally infected cattle, surface aggregation of IgM and CD5 molecules on B-lymphocytes is absent. It is known that the wave-like nature of IgM synthesis, which was shown in previous studies, depends on a subpopulation of B1 cells. After 7 days of the immune response, IgM+ and CD5+ cells do not correlate, which shows their functional difference. The increase in CD5+ cells is probably not associated with B cells, but with T cells differentiating under the influence of the virus. A subset of B1 cells is the primary target of cattle leukemia virus. The 65th day of the immune response is characterized by the expansion of IgM+ B cells, a decrease in the number of CD5+ cells and a uniform distribution of receptors around the perimeter of the cells.


Autophagy Modulators and Neuroinflammation.
Neuroinflammation plays a critical role in the development and progression of various neurological disorders. Therefore, various studies have focused on the development of neuroinflammation inhibitors as potential therapeutic tools. Recently, the involvement of autophagy in the regulation of neuroinflammation has drawn substantial scientific interest, and a growing number of studies support the role of impaired autophagy in the pathogenesis of common neurodegenerative disorders. The purpose of this article is to review recent research on the role of autophagy in controlling neuroinflammation. We focus on studies employing both mammalian cells and animal models to evaluate the ability of different autophagic modulators to regulate neuroinflammation. We have mostly reviewed recent studies reporting anti-neuroinflammatory properties of autophagy. We also briefly discussed a few studies showing that autophagy modulators activate neuroinflammation in certain conditions. Recent studies report neuroprotective as well as anti-neuroinflammatory effects of autophagic modulators. We discuss the possible underlying mechanisms of action of these drugs and their potential limitations as therapeutic agents against neurological disorders. Autophagy activators are promising compounds for the treatment of neurological disorders involving neuroinflammation.


Interprofessional Shared Decision-Making in the ICU: A Systematic Review and Recommendations From an Expert Panel.
There is growing recognition that high-quality care for patients and families in the ICU requires exemplary interprofessional collaboration and communication. One important aspect is how the ICU team makes complex decisions. However, no recommendations have been published on interprofessional shared decision-making. The aim of this project is to use systematic review and normative analysis by experts to examine existing evidence regarding interprofessional shared decision-making, describe its principles and provide ICU clinicians with recommendations regarding its implementation. We conducted a systematic review using MEDLINE, Cumulative Index to Nursing and Allied Health Literature, and Cochrane databases and used normative analyses to formulate recommendations regarding interprofessional shared decision-making. Three authors screened titles and abstracts in duplicate. Four papers assessing the effect of interprofessional shared decision-making on quality of care were identified, suggesting that interprofessional shared decision-making is associated with improved processes and outcomes. Five recommendations, largely based on expert opinion, were developed: 1) interprofessional shared decision-making is a collaborative process among clinicians that allows for shared decisions regarding important treatment questions; 2) clinicians should consider engaging in interprofessional shared decision-making to promote the most appropriate and balanced decisions; 3) clinicians and hospitals should implement strategies to foster an ICU climate oriented toward interprofessional shared decision-making; 4) clinicians implementing interprofessional shared decision-making should consider incorporating a structured approach; and 5) further studies are needed to evaluate and improve the quality of interprofessional shared decision-making in ICUs. Clinicians should consider an interprofessional shared decision-making model that allows for the exchange of information, deliberation, and joint attainment of important treatment decisions.


Indirect genetic effects from competition in the clonal herb Sedum album (Crassulaceae).
Recent years have seen increasing interest in indirect genetic effects, i.e. influences on the phenotype that depend on the genotype of other conspecific individuals; however, the empirical evidence for such effects is still limited, especially in wild plant species. The present study of the clonal herb Sedum album assessed direct and indirect genetic effects on performance-related traits in a 4-year experiment with clonally replicated genotypes, grown in pairs and differing in anthocyanin pigmentation to allow separation of individuals during data collection. In agreement with the existence of indirect genetic effects, the experimentally-paired plants not only expressed their own genotype but were also affected by the genotype of their pair mate. The effect of neighbour genotype explained up to one-fourth of the variation in performance and most likely resulted from competition, imposed by the close physical contact between paired individuals and the limiting conditions used in the garden environment. Indirect genetic effects from competition have the potential to enhance the efficacy of group-level selection relative to individual selection, given the nutrient-poor and spatially-confined substrate available to plants of S. album in the natural habitat.


Role of Monovalent Ions in the NKCC1 Inhibition Mechanism Revealed through Molecular Simulations.
The secondary active Na-K-Cl cotransporter 1 (NKCC1) promotes electroneutral uptake of two chloride ions, one sodium ion and one potassium ion. NKCC1 regulates Cl- homeostasis, thus being implicated in transepithelial water transport and in neuronal excitability. Aberrant NKCC1 transport is linked to a variety of human diseases. The loop diuretic drugs bumetanide, furosemide, azosemide and ethacrynic acid target NKCC1, but are characterized by poor selectivity leading to severe side effects. Despite its therapeutic importance, the molecular details of the NKCC1 inhibition mechanism remain unclear. Using all-atom simulations, we predict a putative binding mode of these drugs to the zebrafish (z) and human (h) NKCC1 orthologs. Although differing in their specific interactions with NKCC1 and/or monovalent ions, all drugs can fit within the same cavity and engage in hydrophobic interactions with M304/M382 in z/hNKCC1, a proposed ion gating residue demonstrated to be key for bumetanide binding. Consistent with experimental evidence, all drugs take advantage of the K+/Na+ ions, which plastically respond to their binding. This study not only provides atomic-level insights useful for drug discovery campaigns of more selective/potent NKCC1 inhibitors aimed to tackle diseases related to deregulated Cl- homeostasis, but it also supplies a paradigmatic example of the key importance of dynamical effects when drug binding is mediated by monovalent ions.


The role of pars plana vitrectomy in the management of fungal endogenous endophthalmitis.
To identify the causative microorganism of fungal endogenous endophthalmitis in our tertiary referral uveitis center and review the therapeutic role of pars plana vitrectomy in patients with fungal endogenous endophthalmitis. Seven eyes of six cases were identified as fungal endogenous endophthalmitis through positive cultures of ocular fluids and clinical presentations. The final anatomical and functional results were evaluated. Four women (66.7%) and two men (33.3%) underwent vitrectomy. Control of infection was achieved early on in all cases. Candida (71.4%) and Aspergillus (28.6%) species were identified as causative fungi in patients with fungal endogenous endophthalmitis. Two patients were reoperated due to reinfection and retinal detachment, respectively. Visual acuity improved in six eyes (85.7%) and worsened in one eye (14.3%). At the final examination, the retina was flat in all cases. No eye developed phthisis bulbi. Candida species are the most common causative organisms of fungal endogenous endophthalmitis in this study. Pars plana vitrectomy in fungal endogenous endophthalmitis may enhance the treatment of infection by removing fungal elements in the vitreous and aid in diagnosis. Vitrectomy may also be an important tool in the management of vision-threatening post-infectious sequelae such as retinal detachment and reinfections.


E5 and E6/E7 of high-risk HPVs cooperate to enhance cancer progression through EMT initiation.
It is estimated that 10-20% of human carcinogenesis is linked to virus infection including papillomaviruses (HPVs). Moreover, since metastatic cancer disease is a major cause of morbidity and mortality in cancer patients, the role of onco-viruses in cancer progression to a metastatic form is of particular interest. Recent studies reported that E5 and E6/E7 onco-proteins of high-risk HPVs could enhance cancer progression via the initiation of the epithelial-mesenchymal transition (EMT) event. Herein, we discuss the association between E5 as well as E6/E7 of high-risk HPV and cancer progression.


Scaling factors to relate drug metabolic clearance in hepatic microsomes, isolated hepatocytes, and the intact liver: studies with induced livers involving diazepam.
Microsomal protein recovery and hepatocellularity have been determined and investigated as scaling factors for interrelating clearance by hepatic microsomes, freshly isolated hepatocytes and whole liver from untreated (UT) rats and rats treated with either the cytochrome P450 inducer phenobarbital (PB) or dexamethasone (DEX). Hepatocellularity in UT rats (1.1 x 10(8) hepatocytes/g liver) was not significantly different after either PB or DEX induction (1.1 and 1.3 x 10(8) hepatocytes/g liver, respectively). However the microsomal protein recovery index, which provides a scaling factor that is inversely related to the efficiency of the microsomal preparation procedure, was 47 mg/g liver in both PB and DEX microsomes and differs from UT rats (60 mg/g liver). These contrasting findings are consistent with the interlaboratory trends in the literature, indicating that, although hepatocellularity estimates are in good accord, microsomal recovery can vary 2-fold; this has implications for scaling. The oxidation of diazepam to its three primary metabolites was measured in PB and DEX microsomes and hepatocytes and the scaling factors were applied to these data and previously reported UT data. Marked changes in kinetics occur on induction resulting in a shift in the major pathway. In particular, 3-hydroxylation is induced over 20-fold by DEX. Diazepam CL(int) was determined in vivo after administration of a bolus dose into the hepatic portal vein of UT, PB, and DEX rats; values of 127, 191, and 323 ml/min/SRW (where SRW is a standard rat weight of 250 g), respectively, were obtained. Using these scaling factors, the hepatocyte predictions of CL(int) were excellent (99, 144, and 297 ml/min/SRW for UT, PB, and DEX, respectively), whereas only the DEX prediction (248 ml/min/SRW) was accurate for the microsomal system, with a substantial underprediction for UT and PB (46 and 68 ml/min/SRW, respectively). Evidence is presented for product inhibition, resulting from accumulation of primary metabolites within the microsomal preparation, as the mechanism responsible for this underprediction. These results illustrate that the scaling factor approach is applicable to induced livers in which both cytochrome P450 complement and zonal distribution are altered. These data, together with our previous studies, demonstrate that CL(int) in cells (2.4-297 ml/min/SRW), microsomes (2.7-248 ml/min/SRW), and in vivo (1.5-323 ml/min/SRW) are related in a linear fashion and hence inherently both in vitro systems are of equal value in predicting in vivo CL(int).


Molecular epidemiology and drug sensitivity pattern of Mycobacterium tuberculosis strains isolated from pulmonary tuberculosis patients in and around Ambo Town, Central Ethiopia.
Tuberculosis (TB) is caused by M. tuberculosis complex and remains a major global public health problem. The epidemic remains a threat to sub-Saharan Africa, including Ethiopia, with further emergence of drug resistant TB. We investigated the drug sensitivity pattern and molecular epidemiology of mycobacterial strains isolated from pulmonary TB patients in and around Ambo town in Oromia Region, Central Ethiopia. A cross-sectional study was conducted involving 105 consecutive new smear positive pulmonary TB patients diagnosed at Ambo Hospital and surrounding Health Centers between May 2014 and March 2015 upon informed consent. Sputum samples were cultured on Löwenstein-Jensen (LJ) media using standard techniques to isolate mycobacteria. Region of difference 9 (RD9)-based polymerase chain reaction (PCR) and spoligotyping was employed for the identification of the isolates at species and strain levels. The spoligotype patterns were entered into the SITVIT database to determine Octal and SIT (Spoligotyping International Typing) numbers for each strain. The sensitivity of the isolates to isoniazid (INH), rifampicin (RIF), ethambutol (ETB) and streptomycin (STM) was evaluated on LJ-medium with the indirect proportion method. Cultures were positive in 86/105 (82%) of newly diagnosed smear positive pulmonary TB cases. All of the 86 isolates were confirmed as M. tuberculosis. The majority (76.7%) of them were clustered into seven groups while the rest (23.3%) appeared unique. The most predominant Spoligotypes were SIT53 and SIT149, consisting of 24.4% and 20.9% of the isolates, respectively. Assigning of the isolates to family using SPOTCLUST software revealed that 45.3% of the isolates belonged to T1, 23.3% to T3 and 13% to CAS family. The majority (76.7%) of the M. tuberculosis isolates were susceptible to all the four drugs. Any resistance to any one of the four drugs was detected in 23.3% of the isolates. The highest proportion of any resistance was observed against isoniazid (9.3%) and ethambutol (7%). There was only a single case (1.2%) of multidrug resistant/rifampicin resistant (MDR/RR) TB. The majority of the isolates were clustered suggesting on-going active transmission in the study area. Mono resistance is relatively prevalent while the magnitude of MDR/RR-TB was found to be lower than in previous studies.


Three-dimensional distributions of elements in biological samples by energy-filtered electron tomography.
By combining electron tomography with energy-filtered electron microscopy, we have shown the feasibility of determining the three-dimensional distributions of phosphorus in biological specimens. Thin sections of the nematode, Caenorhabditis elegans were prepared by high-pressure freezing, freeze-substitution and plastic embedding. Images were recorded at energy losses above and below the phosphorus L2,3 edge using a post-column imaging filter operating at a beam energy of 120 keV. The unstained specimens exhibited minimal contrast in bright-field images. After it was determined that the specimen was sufficiently thin to allow two-window ratio imaging of phosphorus, pairs of pre-edge and post-edge images were acquired in series over a tilt range of +/-55 degrees at 5 degrees increments for two orthogonal tilt axes. The projected phosphorus distributions were aligned using the pre-edge images that contained inelastic contrast from colloidal gold particles deposited on the specimen surface. A reconstruction and surface rendering of the phosphorus distribution clearly revealed features 15-20 nm in diameter, which were identified as ribosomes distributed along the stacked membranes of endoplasmic reticulum and in the cytoplasm. The sensitivity of the technique was estimated at < 35 phosphorus atoms per voxel based on the known total ribosomal phosphorus content of approximately 7000 atoms. Although a high electron dose of approximately 10(7)e/nm2 was required to record two-axis tilt series, specimens were sufficiently stable to allow image alignment and tomographic reconstruction.


Management of uterine hyperstimulation with concomitant use of oxytocin and terbutaline.
The purpose of this study was to evaluate the efficacy of the concomitant use of subcutaneous terbutaline and oxytocin for the management of uterine hyperstimulation. Patients in active labor receiving intravenous oxytocin who developed uterine hyperstimulation were randomly assigned to receive either oxytocin discontinuation or administration of subcutaneous terbutaline while maintaining the oxytocin infusion. Time to resolution of hyperstimulation was the primary end point. Twenty-nine patients were enrolled. The most frequent types of hyperstimulation were tachysystole (26%) and mixed patterns (26%). Resolution time was significantly shorter in the combination therapy versus control ( P = 0.002). Persistence of hyperstimulation 15 minutes after intervention was seen in 53% of the women in the control group versus 0% of the women in the study group. No significant differences were noted in the Apgar scores, rates of cesarean sections, or chorioamnionitis. In the management of uterine hyperstimulation, subcutaneous terbutaline without discontinuation of oxytocin is more effective than the traditional approach of oxytocin discontinuation.


A comparison between the effects of cold exposure in vivo and of noradrenaline in vitro on the metabolism of the brown fat of new-born rabbits.
1. Exposure of new-born rabbits to the cold leads to an increase in the incorporation of [(14)C]glucose into the glycerol of brown-fat triglyceride, but has no effect on [(14)C]glucose incorporation into triglyceride of white fat or liver. The effect of cold exposure on brown-fat triglyceride is abolished by cutting the cervical sympathetic nerve. 2. Brown fat incorporates very little [(14)C]glucose into triglyceride fatty acids, either in vivo or in vitro. 3. Noradrenaline added to incubations of brown fat from new-born rabbits stimulates O(2) consumption, CO(2) output and incorporation of glucose into triglyceride glycerol. The effects of noradrenaline in vitro are therefore consistent with the hypothesis that noradrenaline mediates the response of the brown fat of new-born rabbits to cold exposure. 4. Glycerokinase is present in the brown fat of new-born rabbits, but its activity is much less than that of the glycerokinase in the brown fat of adult rats. 5. Insulin has no effect on O(2) consumption, CO(2) output or glucose uptake in brown fat of new-born rabbits. 6. It is concluded that the thermogenic response of new-born rabbits to cold exposure is accompanied by a selective acceleration of the triglyceride cycle in brown fat. However, resynthesis of triglyceride would not account for more than 1% of the O(2) consumed in vitro by new-born rabbit brown fat in the presence of noradrenaline if respiration remains coupled to phosphorylation.


Extracting data from graphs: A case-study on animal research with implications for meta-analyses.
Systematic reviews with meta-analyses are powerful tools that can answer research questions based on data from published studies. Ideally, all relevant data is directly available in the text or tables, but often it is only presented in graphs. In those cases, the data can be extracted from graphs, but this potentially introduces errors. Here, we investigate to what extent the extracted outcome and error values differ from the original data and if these differences could affect the results of a meta-analysis. Six extractors extracted 36 outcome values and corresponding errors from 22 articles. Differences between extractors were compared using overall concordance correlation coefficients (OCCC), differences between the original and extracted data were compared using concordance correlation coefficients (CCC). To test the possible influence on meta-analyses, random-effects meta-analyses on mean difference comparing original and extracted data were performed. The OCCCs and CCCs were high for both outcome values and errors, CCCs were >0.99 for the outcome and >0.92 for errors. The meta-analyses showed that the overall effect on outcome was very small (median: 0.025, interquartile range: 0.016-0.046). Therefore, data extraction from graphs is a good method to harvest data if it is not provided in the text or tables, and the original authors cannot provide the data.


Osteoprotegerin deficiency attenuates strontium-mediated inhibition of osteoclastogenesis and bone resorption.
Strontium (Sr) exerts an anabolic and antiresorptive effect on bone, but the mechanism remains unknown. Osteoprotegerin (OPG) expressed by osteoblasts plays an important role in regulating bone homeostasis by inhibiting osteoclastogenesis and bone resorption. This study aims at evaluating the role of OPG in Sr-mediated inhibition of osteoclastogenesis and bone resorption. Six-week-old Opg knockout (KO) male mice and their wild-type (WT) littermates were treated orally with vehicle (Veh) or Sr compound (4 mmol/kg) daily for 8 weeks. Bone mass and microstructure in the lumbar spine (L(4)) and proximal tibia were analyzed with micro-computed tomography (µCT). Bone remodeling was evaluated with serum biochemical analysis and static and dynamic bone histomorphometry. Osteoclast differentiation potential and gene expression were analyzed in bone marrow cells. The findings demonstrate that Sr compound treatment results in greater bone volume and trabecular number than Veh treatment in WT mice. The anabolic response of trabecular bone to Sr treatment is attenuated in KO mice. Although Sr treatment significantly decreases in vitro osteoclastogenesis and bone resorption in WT mice, these effects are attenuated in KO mice. Furthermore, Sr treatment profoundly increases Opg gene expression in the tibias and OPG protein levels in the sera of WT mice. This study concludes that the inhibition of osteoclastogenesis and bone resorption is possibly associated with OPG upregulation by Sr treatment.


Differential regulation of insulin-like growth factor binding protein secretion from human decidual cells by IGF-I, insulin, and relaxin.
Several growth hormone-independent 25-31,000 kD insulin-like growth factor binding proteins (IGF-BPs) have been identified in plasma, extravascular fluids, and various cell-conditioned media. Cultured human decidual cells release three IGF-BPs with 24,000, 30,000, and 34,000 Mr. Using ligand blot analysis and an RIA for the 30,000-Mr form (IGF-BP-1), we examined the effects of IGF-I (10-1,000 ng/ml), insulin (10-10,000 ng/ml), and relaxin (10-250 ng/ml) on decidual cell IGF-BP release after 120 h of hormone exposure. IGF-I inhibited release of both IGF-BP-1 and the 24,000 Mr form. Inhibition of IGF-BP-1 release was noted after 48 h of treatment and was progressive throughout the subsequent 120 h. Insulin stimulated a fourfold increase in release of the 24,000-Mr protein while inhibiting IGF-BP-1 release comparable to IGF-I, alpha-IR3, a monoclonal antibody to the IGF-I receptor, blocked approximately 33% of the IGF-I response but had no effect on insulin-mediated IGF-BP-1 inhibition. Relaxin stimulated a 2.4-fold increase in release of the 24,000-Mr form and a 16-fold increase in the 30,000-Mr protein after 120 h. Stimulation of the 30,000-Mr protein was inhibited by the addition of cycloheximide (50 micrograms/ml). Both IGF-I and insulin also blocked the relaxin-mediated increase in IGF-BP-1. These studies suggest that three structurally related proteins differentially regulate IGF-BP secretion possibly via activation of distinct receptor subtypes.


Protein synthesis and neurotrophin-dependent structural plasticity of single dendritic spines.
Long-term potentiation (LTP) at glutamatergic synapses is considered to underlie learning and memory and is associated with the enlargement of dendritic spines. Because the consolidation of memory and LTP require protein synthesis, it is important to clarify how protein synthesis affects spine enlargement. In rat brain slices, the repetitive pairing of postsynaptic spikes and two-photon uncaging of glutamate at single spines (a spike-timing protocol) produced both immediate and gradual phases of spine enlargement in CA1 pyramidal neurons. The gradual enlargement was strongly dependent on protein synthesis and brain-derived neurotrophic factor (BDNF) action, often associated with spine twitching, and was induced specifically at the spines that were immediately enlarged by the synaptic stimulation. Thus, this spike-timing protocol is an efficient trigger for BDNF secretion and induces protein synthesis-dependent long-term enlargement at the level of single spines.


Microbiota, diet, and the gut-brain axis in multiple sclerosis and stroke.
Intestinal microbiota can influence the phenotype and function of immune cell responses through the dissemination of bacterial antigens or metabolites. Diet is one of the major forces shaping the microbiota composition and metabolism, contributing to host homeostasis and disease susceptibility. Currently, nutrition is a complementary and alternative approach to the management of metabolic and neurological diseases and cancer. However, the knowledge of the exact mechanism of action of diet and microbiota on the gut-brain communication is only developing in recent years. Here, we reviewed the current knowledge on the effect of diet and microbiota on the gut-brain axis in patients with two different central nervous system diseases, multiple sclerosis and stroke. We have also highlighted the open questions in the field that we believe are important to address to gain a deeper understanding of the mechanisms by which diet can directly or indirectly affect the host via the microbiota. We think this will open up new approaches to the treatment, diagnosis, and monitoring of various diseases.


Plain-language summaries: An essential component to promote knowledge translation.
In this era of evidence-based practice, scholarly work such as peer-reviewed scientific publications plays a vital role in policy and decision-making at an individual, organisation and country-level. Alongside being considered an essential means of communicating scholarly work, scientific publications also investigate the specific domains that lack well-established literature and thereby inform scientists and researchers to thrive for the betterment of the publics' well-being. Thus, the main purpose of articulating the scientific, scholarly work should be to make it understandable and accessible to everyone, including the lay audience. However, oftentimes, researchers overlook the lay summaries while publishing the research findings.


Alcohol intake and cigarette smoking and risk of a contralateral breast cancer: The Women's Environmental Cancer and Radiation Epidemiology Study.
Women with primary breast cancer are at increased risk of developing second primary breast cancer. Few studies have evaluated risk factors for the development of asynchronous contralateral breast cancer in women with breast cancer. In the Women's Environmental Cancer and Radiation Epidemiology Study (1985-2001), the roles of alcohol and smoking were examined in 708 women with asynchronous contralateral breast cancer (cases) compared with 1,399 women with unilateral breast cancer (controls). Cases and controls aged less than 55 years at first breast cancer diagnosis were identified from 5 population-based cancer registries in the United States and Denmark. Controls were matched to cases on birth year, diagnosis year, registry region, and race and countermatched on radiation treatment. Risk factor information was collected by telephone interview. Rate ratios and 95% confidence intervals were estimated by using conditional logistic regression. Ever regular drinking was associated with an increased risk of asynchronous contralateral breast cancer (rate ratio = 1.3, 95% confidence interval: 1.0, 1.6), and the risk increased with increasing duration (P = 0.03). Smoking was not related to asynchronous contralateral breast cancer. In this, the largest study of asynchronous contralateral breast cancer to date, alcohol is a risk factor for the disease, as it is for a first primary breast cancer.


A mutation in the expansin-like A2 gene enhances resistance to necrotrophic fungi and hypersensitivity to abiotic stress in Arabidopsis thaliana.
Expansins are cell wall loosening agents, known for their endogenous function in cell wall extensibility. The Arabidopsis expansin-like A2 (EXLA2) gene was identified by its down-regulation in response to infection by the necrotrophic pathogen Botrytis cinerea, and by the reduced susceptibility of an exla2 mutant to the same pathogen. The exla2 mutant was equally susceptible to Pseudomonas syringae pv. tomato, but was more resistant to the necrotrophic fungus Alternaria brassicicola, when compared with the wild-type or with transgenic, ectopic EXLA2-overexpressing lines. The exla2 mutants also enhanced tolerance to the phytoprostane-A1 . This suggests that the absence or down-regulation of EXLA2 leads to increased resistance to B. cinerea in a CORONATINE INSENSITIVE 1 (COI1)-dependent manner, and this down-regulation can be achieved by phytoprostane-A1 treatment. EXLA2 is induced significantly by salinity and cold, and by the exogenous application of abscisic acid. The exla2 mutant also showed hypersensitivity towards increased salt and cold, and this hypersensitivity required a functional abscisic acid pathway. The differential temporal expression of EXLA2 and the phenotypes in transgenic plants with altered expression of EXLA2 indicate that plant cell wall structure is an important player during Arabidopsis developmental stages. Our results indicate that EXLA2 appears to be important in response to various biotic and abiotic stresses, particularly in the pathogenesis of necrotrophic pathogens and in the tolerance to abiotic stress.


WatershedCounting3D: a new method for segmenting and counting punctate structures from confocal image data.
Current research in cell biology frequently uses light microscopy to study intracellular organelles. To segment and count organelles, most investigators have used a global thresholding method, which relies on homogeneous background intensity values within a cell. Because this is not always the case, we developed WatershedCounting3D, a program that uses a modified watershed algorithm to more accurately identify intracellular structures from confocal image data, even in the presence of an inhomogeneous background. We give examples of segmenting and counting endoplasmic reticulum exit sites and the Golgi apparatus.


Alginate Microsphere Encapsulation of Drug-Loaded Nanoparticles: A Novel Strategy for Intraperitoneal Drug Delivery.
Alginate hydrogels have been broadly investigated for use in medical applications due to their biocompatibility and the possibility to encapsulate cells, proteins, and drugs. In the treatment of peritoneal metastasis, rapid drug clearance from the peritoneal cavity is a major challenge. Aiming to delay drug absorption and reduce toxic side effects, cabazitaxel (CAB)-loaded poly(alkyl cyanoacrylate) (PACA) nanoparticles were encapsulated in alginate microspheres. The PACAlg alginate microspheres were synthesized by electrostatic droplet generation and the physicochemical properties, stability, drug release kinetics, and mesothelial cytotoxicity were analyzed before biodistribution and therapeutic efficacy were studied in mice. The 450 µm microspheres were stable at in vivo conditions for at least 21 days after intraperitoneal implantation in mice, and distributed evenly throughout the peritoneal cavity without aggregation or adhesion. The nanoparticles were stably retained in the alginate microspheres, and nanoparticle toxicity to mesothelial cells was reduced, while the therapeutic efficacy of free CAB was maintained or improved in vivo. Altogether, this work presents the alginate encapsulation of drug-loaded nanoparticles as a promising novel strategy for the treatment of peritoneal metastasis that can improve the therapeutic ratio between toxicity and therapeutic efficacy.


Naturally occurring plasmids in Acinetobacter calcoaceticus: pAV2, a plasmid which influences the fertility of the sex factor pAV1.
Acinetobacter calcoaceticus strain EBF65/65 harbours a cryptic plasmid, pAV2, which has been shown by electrophoretic separation on agarose gels to have a molecular mass of approximately 13.5 megadaltons (Md). Transfer of the previously described sex factor pAV1 (Hinchliffe & Vivian, 1980 a,b) from the hospital strainJC17 into strains possessing pAV2 occurs only at a low frequency, whereas transfer to similar strains lacking pAV2 occurs at a much higher frequency. In EBF65/65, pAV1 may be present in strains possessing or lacking pAV2; pAV1 strains lacking pAV2 correspond to strains previously described as pAV1a (Hinchliffe & Vivian, 1980b) whereas pAV1 strains which also possess pAV2 correspond to pAB1b strains. The genetic evidence presented here is consistent with the hypothesis that pAV2 specifies a host restriction and modification system that is active against pAV1. Physical evidence from agarose gel electrophoresis indicates that pAV1 corresponds to a band of approximately 85 Md in strain JC17. The corresponding band in strains of EBF65/65 is difficult to distinguish because of the presence of a further cryptic plasmid band of approximately 88 Md, designated pAV3. A small cryptic plasmid of approximately 6 Md, designated pAV4, is reported for EBF65/65.


Animal Health Ireland: providing national leadership and coordination of non-regulatory animal health issues in Ireland.
Livestock production plays an important role in the Irish economy. Regulatory animal health issues are the responsibility of government, but until recently there has been no national coordination of non-regulatory animal health issues. This gap has recently been filled with the establishment of Animal Health Ireland (AHI), a not-for-profit, partnership-based organisation providing national leadership and coordination of non-regulatory animal health issues in Ireland. Animal Health Ireland provides benefits to livestock producers and processors by providing the knowledge, education and coordination required to establish effective control strategies, both on-farm and nationally. This paper presents a brief overview of the context for AHI, and of its establishment and initial activities. Non-regulatory animal health issues have been prioritised. A series of work programmes (each focusing on a high-priority issue) have been established. Partnership is critical to success, both for AHI as an organisation and for effective farm-level transfer of knowledge. This model for national leadership and coordination of non-regulatory animal health issues may be of relevance elsewhere.


Worldwide trends in asthma mortality.
Statistics for deaths from asthma yield widely variable mortality rates between countries. Validation studies show significant inaccuracies in certification of cause of death in older subjects, and probable underreporting of asthma deaths in some countries, explaining some of the international differences noted. The introduction of the ninth revision of the World Health Organisation International Classification of Diseases brought a step increase in reported asthma mortality rates in 1979, but the last decade has seen a gradual increase in mortality rates in many countries, especially in young people, not explained by this change in code. Diagnostic transfer does not adequately explain these increases. Individuals at risk of death from asthma are more likely to be non-caucasian, non-compliant, and young, with a history of previous life-threatening episodes, hospital admissions, and precipitous attacks. Studies of circumstances of death have emphasized overreliance on beta-agonists and underuse of corticosteroids as two primary deficiencies in management, but there remains debate about the causality of the association between increased asthma mortality and increased usage of beta-agonists. The gradual increase in asthma mortality seen in New Zealand over the last 40 years, with episodic increases in the rising baseline, together with similar but more gradual changes in other countries, raises concerns about whether current treatment practices may adversely affect asthma severity and mortality. Further studies are required to validate asthma mortality statistics, and to establish causation of deaths.


Video-assisted anal fistula treatment: a high volume unit initial experience.
Introduction Perianal fistulas are a common problem. Video-assisted anal fistula treatment is a new technique for the management of this difficult condition. We describe our initial experience with the technique to facilitate the treatment of established perianal fistulas. Methods We reviewed a prospectively maintained database relating to consecutive patients undergoing video-assisted anal fistula treatment in a single unit. Results Seventy-eight consecutive patients had their perianal fistulas treated with video-assistance from November 2014 to June 2016. Complete follow-up data were available in 74 patients, with median follow-up of 14 months (interquartile range 7-19 months). There were no complications and all patients were treated as day cases. Most patients had recurrent disease, with 57 (77%) having had previous fistula surgery. At follow-up, 60 (81%) patients reported themselves 'cured' (asymptomatic) including 5 patients with Crohn's disease and one who had undergone 10 previous surgical procedures. Logistical stepwise regression did not demonstrate any statistically significant factors that may have been considered to affect outcome (age, gender, diabetes, previous I&D, Crohn's disease, smoking, type of fistula). Conclusions Our data have shown that video-assisted anal fistula treatment is safe and effective in the management of perianal fistulas in our patients and this suggests it may be applied to all patients regardless of comorbidity, underlying pathology or type of fistula.


Impact of social standing on injury prevention in a World Health Organization Safe Community--intervention outcome by household employment contract.
Although social inequality in health has been an argument for community-based injury prevention programmes, intervention outcomes with regard to differences in social standing have not been analysed. The objective of this study was to investigate rates of injuries treated in health-care among members of households at different levels of labour market integration before and after the implementation of a WHO Safe Community programme. A quasi-experimental design was used with pre- and post-implementation data collection covering the total populations <65 years of age during one year in the programme implementation municipality (population 41 000) and in a control municipality (population 26 000). Changes in injury rates were studied using prospective registration of all acute care episodes with regard to social standing in both areas during the study periods. Male members of households categorized as not vocationally active displayed the highest pre-intervention injury rates. Also after the intervention, males in households classified as not vocationally active displayed notably elevated injury rates in both the control and study areas. Households in the study area in which the significant member was employed showed a post-intervention decrease in injury rate among both men (P < 0.001) and women (P < 0.01). No statistically significant change was observed in households in which the significant member was self-employed or not vocationally active. In the control area, only an aggregate-level decrease (P < 0.05) among members of households in which the significant member was employed was observed. The study displayed areas for improvement in the civic network-based WHO Safe Community model. Even though members of non-vocationally active households, in particular men, were at higher pre-intervention injury risk, they were not affected by the interventions. This fact has to be addressed when planning future community-based injury prevention programmes.


Efficacy of Oral Furosemide Test for Primary Aldosteronism Diagnosis.
Confirmatory tests represent a fundamental step in primary aldosteronism (PA) diagnosis, but they are laborious and often require a hospital environment due to the risks involved. To evaluate the efficacy of oral furosemide as a new confirmatory test for PA diagnosis. We prospectively evaluated the diagnostic performance of 80 mg of oral furosemide in 64 patients with PA and 22 with primary hypertension (controls). Direct renin concentration (DRC) was measured before, and 2 hours and 3 hours after the oral furosemide. In addition, the oral furosemide test was compared with 2 other confirmatory tests: the furosemide upright test (FUT) and saline infusion test (SIT) or captopril challenge test (CCT) in all patients with PA. The cut-off of 7.6 µU/mL for DRC at 2 hours after oral furosemide had a sensitivity of 92%, specificity of 82%, and accuracy of 90% for PA diagnosis. In 5 out of 6 controls with low-renin hypertension, which might represent a PA spectrum, renin remained suppressed. Excluding these 6 controls with low-renin hypertension, the DRC cut-off of 10 µU/mL at 2 hours after oral furosemide had a sensitivity of 95.3%, specificity of 93.7% and accuracy of 95% for PA diagnosis. DRC after 3 hours of oral furosemide did not improve diagnostic performance. Using the cut-off of 10 µU/mL, the oral furosemide test and the FUT were concordant in 62 out of 64 (97%) patients with PA. Only 4 out of 64 cases with PA (6.4%) ended the oral furosemide test with potassium <3.5 mEq/L. Hypotension was not evidenced in any patient with PA during the test. The oral furosemide test was safe, well-tolerated and represents an effective strategy for PA investigation.


[Emetic and antiemetic properties of regulatory peptides].
Emetic and antiemetic properties of opioid peptides, substance "P", beta-lipotropin, and ACTH1-39 have been investigated in experiments on cats. It was shown that morphine, enkephalin, beta-endorphin and DADLE caused vomiting in animals, which was blocked by naloxone. Substance "P", gamma- and des-tyr-gamma-endorphin manifested antiemetic properties similar to those of naloxone. Selective antagonists of delta-opioid receptors ICI 154, 129 blocked emetic action of delta-agonist DADLE but did not prevent vomiting caused by mu-agonist morphine. It is suggested that the vomiting mechanisms of endogenous opioid peptides involve stimulation of mu- and delta-opioid receptors in the chemoreceptor trigger zone of the vomiting centre.


Long-term results of transanal excision after neoadjuvant chemoradiation for T2 and T3 adenocarcinomas of the rectum.
Traditionally, selected early distal rectal cancers have been considered for treatment by transanal excision (TAE) with acceptable oncologic results. With the frequent use of neoadjuvant chemoradiation (NCR) for the treatment of locally advanced rectal cancer, there is growing interest in the application of TAE for such lesions. We report our experience of TAE for T2 and T3 rectal cancers following NCR. Between July 1994 and August 2006, 44 patients were identified as having undergone full-thickness TAE of pretreatment ultrasound-staged T2 and T3 rectal cancers that were treated with NCR. Fifteen patients were deemed medically unfit for radical resection, and 29 would have required abdominoperineal resection but were opposed to colostomy. Our patient population consisted of 26 men and 18 women, with a median age of 69 (range, 43-89) and a median follow up of 64 months (6-153). Thirty-one patients had a clinical complete response (cCR) to NCR of which 19 (61%) had a pathologic CR (pCR). Seven (16%) of 44 patients sustained disease recurrence of which two were local only, two local and systemic, and three systemic only. Only four (9%) patients had died of disease at current follow up. Overall 5-year survival rates for T2/T3N0 and T2/T3N1 patients were 84% and 81%, respectively. Five patients underwent radical resection immediately following TAE for either positive margins or residual cancer. There was minimal morbidity with no perioperative mortality associated with TAE. TAE of T2 and T3 rectal cancers following NCR is a safe alternative to radical resection in a highly select group of patients for which recurrence and survival rates comparable to radical resection can be achieved. This study supports ongoing efforts to assess this approach in prospective, multi-center trials.


Voluntary and involuntary maternal separation in guinea pig pups with mothers required to forage.
Lactating guinea pigs required to leave a nest box to obtain food were observed to be separated from their pups almost 10% of the time on Days 13-15 postpartum. The duration of separation was greater in the dark than in the light, and was not different for mothers/litters exposed to low (LFD) versus high (HFD) foraging demand. Subsequently, LFD and HFD pups as well as pups reared under standard laboratory conditions (SLC) all exhibited vocalization and plasma cortisol responses to involuntary maternal separation. However, several effects of rearing were also observed. HFD pups vocalized less during separation and had higher resting cortisol levels than did SLC pups; LFD and HFD males spent less time in contact with the mother in a test cage than did SLC males. These effects are discussed in terms of the importance of the physical environment in shaping infant-mother interactions and the infant's perception of control.


Studies of Coumarin Derivatives for Constitutive Androstane Receptor (CAR) Activation.
Constitutive androstane receptor (CAR) activation has found to ameliorate diabetes in animal models. However, no CAR agonists are available clinically. Therefore, a safe and effective CAR activator would be an alternative option. In this study, sixty courmarin derivatives either synthesized or purified from Artemisia capillaris were screened for CAR activation activity. Chemical modifications were on position 5,6,7,8 with mono-, di-, tri-, or tetra-substitutions. Among all the compounds subjected for in vitro CAR activation screening, 6,7-diprenoxycoumarin was the most effective and was selected for further preclinical studies. Chemical modification on the 6 position and unsaturated chains were generally beneficial. Electron-withdrawn groups as well as long unsaturated chains were hazardous to the activity. Mechanism of action studies showed that CAR activation of 6,7-diprenoxycoumarin might be through the inhibition of EGFR signaling and upregulating PP2Ac methylation. To sum up, modification mimicking natural occurring coumarins shed light on CAR studies and the established screening system provides a rapid method for the discovery and development of CAR activators. In addition, one CAR activator, scoparone, did showed anti-diabetes effect in db/db mice without elevation of insulin levels.


TADB 3.0: an updated database of bacterial toxin-antitoxin loci and associated mobile genetic elements.
TADB 3.0 (https://bioinfo-mml.sjtu.edu.cn/TADB3/) is an updated database that provides comprehensive information on bacterial types I to VIII toxin-antitoxin (TA) loci. Compared with the previous version, three major improvements are introduced: First, with the aid of text mining and manual curation, it records the details of 536 TA loci with experimental support, including 102, 403, 8, 14, 1, 1, 3 and 4 TA loci of types I to VIII, respectively; Second, by leveraging the upgraded TA prediction tool TAfinder 2.0 with a stringent strategy, TADB 3.0 collects 211 697 putative types I to VIII TA loci predicted in 34 789 completely sequenced prokaryotic genomes, providing researchers with a large-scale dataset for further follow-up analysis and characterization; Third, based on their genomic locations, relationships of 69 019 TA loci and 60 898 mobile genetic elements (MGEs) are visualized by interactive networks accessible through the user-friendly web page. With the recent updates, TADB 3.0 may provide improved in silico support for comprehending the biological roles of TA pairs in prokaryotes and their functional associations with MGEs.


The veterinary medicine industry in Britain in the twentieth century.
Economic historians have focused research effort on accounting for the growth and significance of Britain's pharmaceutical industry, but little effort has so far been directed at the veterinary medicine industry, which formed an important part of the wider sector. This article addresses that gap. Factors responsible for that sector's relative insignificance until the 1950s included a general tendency to slaughter rather than to treat sick animals, the absence of advanced medicines until the innovation of sulpha drugs and antibiotics, and difficult relations with the wider pharmaceutical industry. Thereafter output of veterinary medicines increased dramatically, arising from an exponential growth in the demand for intensively farmed poultry meat. Since the 1980s a decline in the use of drugs in agriculture has caused the industry to concentrate on the health needs of domestic animals rather than those of livestock.


Acceleration of bone development and regeneration through the Wnt/β-catenin signaling pathway in mice heterozygously deficient for GSK-3β.
Glycogen synthase kinase (GSK)-3β plays an important role in osteoblastogenesis by regulating the Wnt/β-catenin signaling pathway. Therefore, we investigated whether GSK-3β deficiency affects bone development and regeneration using mice heterozygously deficient for GSK-3β (GSK-3β(+/-)). The amounts of β-catenin, c-Myc, cyclin D1, and runt-related transcription factor-2 (Runx2) in the bone marrow cells of GSK-3β(+/-) mice were significantly increased compared with those of wild-type mice, indicating that Wnt/β-catenin signals were enhanced in GSK-3β(+/-) mice. Microcomputed tomography of the distal femoral metaphyses demonstrated that the volumes of both the cortical and trabecular bones were increased in GSK-3β(+/-) mice compared with those in wild-type mice. Subsequently, to investigate the effect of GSK-3β deficiency on bone regeneration, we established a partial bone defect in the femur and observed new bone at 14 days after surgery. The volume and mineral density of the new bone were significantly higher in GSK-3β(+/-) mice than those in wild-type mice. These results suggest that bone formation and regeneration in vivo are accelerated by inhibition of GSK-3β, probably through activation of the Wnt/β-catenin signaling pathway.


Individualized prognostic signature for pancreatic carcinoma validated by integrating immune-related gene pairs (IRGPs).
Increasingly attention is being given to immune molecules in pancreatic cancer. The purpose of this study was to understand the potential clinical application of immune-regulated genes (IRGs) in the stratification of prognosis and to facilitate the development of personalized prognostic information for pancreatic cancer patients. We systematically used public data to comprehensively analyze immune-regulated gene pair (IRGP) expression profiles and clinical data. In our study, IRGP signature was identified to predict the overall survival (OS) of pancreatic cancer patients. We suggested that immune genes are enriched in different risk groups. In the high-risk group, M1 macrophages and resting NK cells were significantly enriched, while the percentages of naïve B cells, resting dendritic cells, CD8 T cells and regulatory T cells (Tregs) were significantly higher in the low-risk group, and we verified these results with immunohistochemical experiments. Gene ontology (GO) analysis confirmed that the IRGP index (IRGPI) signature genes in the cohort were mostly party to sensory perception of a chemical stimulus and the adaptive immune response. The identification of these pathways provides a basis for studying the molecular mechanisms of IRGPI signaling to predict the prognosis of pancreatic cancer. Our study effectively constructed a robust IRGP signature with prognostic value for pancreatic cancer, presenting a conceivable method for deciding on a preoperative treatment.


Constructing perfect cubic Ag-Cu alloyed nanoclusters through selective elimination of phosphine ligands.
The aspiration of chemists has always been to design and achieve control over nanoparticle morphology at the atomic level. Here, we report a synthesis strategy and crystal structure of a perfect cubic Ag-Cu alloyed nanocluster, [Ag55Cu8I12(S-C6H32,4(CH3)2)24][(PPh4)] (Ag55Cu8I12 for short). The structure of this cluster was determined by single-crystal X-ray diffraction (SCXRD) and further validated by X-ray photoelectron spectroscopy (XPS), inductively coupled plasma (ICP), Energy-dispersive X-ray spectroscopy (EDX), thermogravimetric analysis (TGA), and 1H and 31P nuclear magnetic resonance (NMR). The surface deviation of the cube was measured to be 0.291 Å, making it the flattest known cube to date.