Matrix and mineral in the sea urchin larval skeleton.
The endoskeletal spicules of sea urchin larvae are composed of calcite, a surrounding extracellular matrix, and small amounts of occluded matrix proteins. The spicules are formed by primary mesenchyme cells (PMCs) in the blastocoel of the embryo, where they adopt stereotypical locations, thereby specifying where spicules will form. PMCs also fuse to form cytoplasmic cords connecting the cell bodies, and it is within the cords that spicules arise. The mineral phase contains 5% Mg as well as Ca, and about 0.1% of the mass is protein. The matrix and mineral form concentric plies, and the composite has different physical properties than those of pure calcite. The calcite diffracts as a single crystal and is composed of well-ordered, but not perfectly ordered, microdomains. There is evidence for adsorption of matrix proteins to specific crystal faces at domain boundaries, which may help regulate crystal growth and texture. Immature spicules contain considerable precipitated amorphous CaCO3, and PMCs also have vesicles that contain amorphous CaCO3. This suggests the hypothesis that the cellular precursor to the spicules is actually amorphous CaCO3 stabilized in the cell by protein. The spicule s enveloped by the PMC cord, but is topologically exterior to the cell. The PMC plasmalemma is tightly applied to the developing spicules, except perhaps at the elongating tip. The characteristics, localization, and possible function of the four identified matrix proteins are discussed. SM50, SM37, and PM27 all primarily enclose the mineral, though small amounts are occluded. SM30 is found in cellular vesicles and is probably the principal occluded protein of the spicule.


Bringing customers into the boardroom.
Misguided marketing strategies have destroyed more shareholder value than shoddy accounting or shady fiscal practices. Yet marketing functions typically reside deep in the organization, far from the executive suite and boardroom, and they are often poorly aligned with corporate strategy. Boards of directors, it would seem, have compelling reasons to monitor their companies' marketing activities. The authors argue that boards lack a clear understanding of how their companies are meeting customers' needs and how their marketing strategies drive (or often fail to drive) top-line growth. To help remedy that problem, they've devised a "marketing dashboard," a series of management reports that could give the board this critical knowledge. The dashboard has three parts, each of which the board should review regularly. The first part tracks the company's main business drivers--those business conditions that, when manipulated or otherwise changed, will directly and predictably affect the company's performance. The second part describes the specific innovations in a pipeline of growth ideas that will allow the company to reach its short- and long-term revenue goals. And the third part provides an overview of the company's marketing skill set so the board can determine not only if the company has enough marketing talent but also if it has the right marketing talent. Unlike isolated measures of marketing performance that are often insufficient, irrelevant, or misleading, the dashboard allows the board to quickly and routinely assess the effectiveness of its company's marketing strategies. Armed with a clear understanding of marketing's role and performance, the board can expose inadequate marketing campaigns, direct management to address the problem, and monitor progress.


Low-grade adipose tissue inflammation in patients with mild-to-moderate chronic obstructive pulmonary disease.
Low-grade systemic inflammation is common in chronic obstructive pulmonary disease (COPD), but its source remains unclear. Adipose tissue is a potent producer of inflammatory mediators and may contribute to systemic inflammation in COPD, possibly via hypoxia. We studied the influence of COPD and exercise-induced oxygen desaturation on adipose tissue inflammation (ATI) and its contribution to systemic inflammation. Subcutaneous adipose tissue biopsies were investigated in 28 clinically stable COPD patients [forced expiratory volume in 1 s: 58 ± 16% predicted; BMI (in kg/m(2)): 24.9 ± 2.9] and 15 age-, sex-, and body composition-matched healthy control subjects. Fat mass was measured with dual-energy X-ray absorptiometry. Patients were prestratified by oxygen desaturation assessed by incremental cycle ergometry. The adipocyte size and adipose tissue expression of 19 inflammatory and hypoxia-related genes were measured, and adipose tissue macrophages (ATMs) were histologically quantified. Systemic inflammatory markers included C-reactive protein (CRP) and a panel of 20 adipokines. COPD patients had comparable fat mass but higher CRP and HOMA-IR than did control subjects. COPD patients and control subjects had comparable adipose tissue gene expression, adipocyte size, ATM infiltration, and systemic adipokine concentrations. Desaturating COPD patients had no different ATI status than did nondesaturating COPD patients. COPD patients with high CRP had significantly greater ATM infiltration than did patients with low CRP, which was independent of BMI and fat mass. In COPD patients, mild-to-moderate COPD, per se, does not enhance ATI or its contribution to systemic inflammation compared with in well-matched healthy control subjects. However, to our knowledge, our study provides a first indication for a possible role of ATMs in the systemic inflammatory response in COPD that requires additional investigation. This trial was registered at www.trialregister.nl as NTR1402.


Bacteria isolated from parasitic nematodes--a potential novel vector of pathogens?
Bacterial pathogens are ubiquitous in soil and water - concurrently so are free-living helminths that feed on bacteria. These helminths fall into two categories; the non-parasitic and the parasitic. The former have been the focus of previous work, finding that bacterial pathogens inside helminths are conferred survival advantages over and above bacteria alone in the environment, and that accidental ingestion of non-parasitic helminths can cause systemic infection in vertebrate hosts. Here, we determine the potential for bacteria to be associated with parasitic helminths. After culturing helminths from fecal samples obtained from livestock the external bacteria were removed. Two-hundred parasitic helminths from three different species were homogenised and the bacteria that were internal to the helminths were isolated and cultured. Eleven different bacterial isolates were found; of which eight were indentified. The bacteria identified included known human and cattle pathogens. We concluded that bacteria of livestock can be isolated in parasitic helminths and that this suggests a mechanism by which bacteria, pathogenic or otherwise, can be transmitted between individuals. The potential for helminths to play a role as pathogen vectors poses a potential livestock and human health risk. Further work is required to assess the epidemiological impact of this finding.


Rare congenital anomalies of the internal carotid artery: anatomic and radiologic aspects of three cases and review of the literature.
Congenital anomaly of the internal carotid artery (ICA) is a rare entity. It is usually discovered incidentally by color doppler carotid sonography, angiography, computerized tomography (CT), or magnetic resonance imaging of the head and neck region taken for some other reasons. The aim of this study was to detect congenital ICA anomalies, to delineate existing collateral vessels and to find out its incidence. 1847 patients' CT angiography images of the head and neck region taken between May 2013 and February 2018 were retrospectively evaluated for ICA anomalies. We detected three cases (0.16%) with unilateral agenesis of ICA, bilateral agenesis of ICA and bilateral hypoplasia of ICA, respectively. Most patients are asymptomatic because of collateral cerebral circulation supplied by the communicating arteries of the circle of Willis, intercavernous anastomosis, communicating arteries from the external carotid artery, and by persistent embryologic arteries to the carotid artery territory. Recognition of ICA anomalies has important implications during planned carotid or transsphenoidal surgery, in thromboembolic disease, and in the follow-up and detection of associated cerebral aneurysms.


Role of transforming growth factor Beta in human cancer.
Transforming growth factor beta (TGF-beta) is a ubiquitous and essential regulator of cellular and physiologic processes including proliferation, differentiation, migration, cell survival, angiogenesis, and immunosurveillance. Alterations in the TGF-beta signaling pathway, including mutation or deletion of members of the signaling pathway and resistance to TGF-beta-mediated inhibition of proliferation are frequently observed in human cancers. Although these alterations define a tumor suppressor role for the TGF-beta pathway in human cancer, TGF-beta also mediates tumor-promoting effects, either through differential effects on tumor and stromal cells or through a fundamental alteration in the TGF-beta responsiveness of the tumor cells themselves. TGF-beta and members of the TGF-beta signaling pathway are being evaluated as prognostic or predictive markers for cancer patients. Ongoing advances in understanding the TGF-beta signaling pathway will enable targeting of this pathway for the chemoprevention and treatment of human cancers.


Supercomputer analysis of sedimentary basins.
Geological processes of fluid transport and chemical reaction in sedimentary basins have formed many of the earth's energy and mineral resources. These processes can be analyzed on natural time and distance scales with the use of supercomputers. Numerical experiments are presented that give insights to the factors controlling subsurface pressures, temperatures, and reactions; the origin of ores; and the distribution and quality of hydrocarbon reservoirs. The results show that numerical analysis combined with stratigraphic, sea level, and plate tectonic histories provides a powerful tool for studying the evolution of sedimentary basins over geologic time.


[Between New Responsibility and Daily Routines - The Role of the VERAH in GP Care: A Qualitative Secondary Data Case Study of the Introduction of Software-Based Case Management].
The concept of Care Assistant in General Practice (VERAH) was developed in order to integrate non-medical staff more strongly into primary care and thus to meet the increasing demand for care and the simultaneous shortage of medical staff. VERAHs are increasingly responsible for software-supported case management and are thus confronted with new tasks. The aim of this study was therefore to explore the role of the VERAH in primary care practices. The present study is a qualitative secondary data analysis; the data collection took place within the projects VESPEERA and TelePraCMan. Twenty individual interviews and two focus group meetings were conducted with a total of 30 physicians, VERAHs and medical assistants from primary care. The data were analysed qualitatively according to Emerson. Contextual and socio-demographic data were collected with an accompanying questionnaire. The VERAHs of all primary care practices from which interview partners participated performed tasks within software-supported case management. Concerning the role of the VERAH, three themes were identified in the interviews: a) concrete tasks of the VERAH in software-supported case management within the practice team, b) relevance of software-supported case management within the activities of the VERAHS and c) relationship between VERAHs and patients. Taking over tasks in software-supported case management can contribute to strengthening and expanding the role of the VERAH. In the future, more attention should be paid to a clear description of the new role, and the conditions of the VERAHs' task fulfilment should be considered.


Anaphylaxis to iodinated contrast material: nonallergic hypersensitivity or IgE-mediated allergy?
Contrast material is generally well tolerated although approximately 1% of patients who receive low-osmolar nonionic contrast material will develop anaphylaxis symptoms. Because most anaphylactic reactions are mild and nonallergic, clinically mimicking immunoglobulin E (IgE)-mediated allergy, diagnostic skin testing has been discussed controversially in the past and prophylactic pretreatment drug regimens are recommended instead. In the past 6 years, all patients with contrast material-induced anaphylaxis have been subjected to allergologic diagnostic procedures to clearly differentiate allergic and nonallergic anaphylaxis. Thus the purpose of our study was to identify and differentiate IgE-mediated allergy and nonallergic contrast material-induced hypersensitivity. Furthermore, the objective of our diagnostic procedures was not only to identify the culprit contrast material but also to find alternative contrast material for future radiologic interventions. We evaluated 96 patients with anaphylaxis symptoms after contrast material application using standardized intradermal skin testing. In patients with positive skin tests, the IgE-mediated allergy was further evaluated with in vitro and challenge tests. In four patients (suffering from anaphylaxis grades 2 and 3) out of the 96 (4.2%), skin tests and basophil activation tests strongly suggested IgE-mediated allergy to the contrast materials iopromide (two patients), iomeprol, and iopentol. In two patients with allergies to iopromide and iomeprol, alternative nonionic monomer contrast materials were tolerated, as identified in controlled challenge tests with iopamidol and iopromide, respectively. The evaluation of patients with contrast material-induced anaphylaxis (at least those with anaphylaxis > or = grade 2) should always include appropriate skin tests ensuring that patients with an IgE-mediated allergy are not missed. Moreover, allergologic testing may identify a contrast material of the group of nonionic monomers that will be tolerated in future radiologic interventions.


Increase in trigeminal ganglion neurons that respond to both calcitonin gene-related peptide and pituitary adenylate cyclase-activating polypeptide in mouse models of chronic migraine and posttraumatic headache.
A large body of animal and human studies indicates that blocking peripheral calcitonin gene-related peptide (CGRP) and pituitary adenylate cyclase-activating polypeptide (PACAP) signaling pathways may prevent migraine episodes and reduce headache frequency. To investigate whether recurring migraine episodes alter the strength of CGRP and PACAP signaling in trigeminal ganglion (TG) neurons, we compared the number of TG neurons that respond to CGRP and to PACAP (CGRP-R and PACAP-R, respectively) under normal and chronic migraine-like conditions. In a mouse model of chronic migraine, repeated nitroglycerin (NTG) administration significantly increased the number of CGRP-R and PACAP-R neurons in TG but not dorsal root ganglia. In TG neurons that express endogenous αCGRP, repeated NTG led to a 7-fold increase in the number of neurons that respond to both CGRP and PACAP (CGRP-R&PACAP-R). Most of these neurons were unmyelinated C-fiber nociceptors. This suggests that a larger fraction of CGRP signaling in TG nociceptors may be mediated through the autocrine mechanism, and the release of endogenous αCGRP can be enhanced by both CGRP and PACAP signaling pathways under chronic migraine condition. The number of CGRP-R&PACAP-R TG neurons was also increased in a mouse model of posttraumatic headache (PTH). Interestingly, low-dose interleukin-2 treatment, which completely reverses chronic migraine-related and PTH-related behaviors in mouse models, also blocked the increase in both CGRP-R and PACAP-R TG neurons. Together, these results suggest that inhibition of both CGRP and PACAP signaling in TG neurons may be more effective in treating chronic migraine and PTH than targeting individual signaling pathways.


Adrenomedullin-(22-52) antagonizes vasodilator responses to CGRP but not adrenomedullin in the cat.
The effects of adrenomedullin (ADM)-(22-52), a putative ADM receptor antagonist, on vasodilator responses to ADM and the structurally related peptide, calcitonin gene-related peptide (CGRP), were investigated in the hindlimb vascular bed of the cat under constant-flow conditions. ADM-(22-52) had no significant effect on hindlimb perfusion pressure when injected in doses up to 120 nmol; after administration of ADM-(22-52), vasodilator responses to ADM were unchanged, whereas vasodilator responses to CGRP were inhibited. The inhibitory effects of ADM-(22-52) on responses to CGRP were selective and reversible and were similar to the inhibitory effects of the CGRP antagonist CGRP-(8-37). Hindlimb vasodilator responses to CGRP and to ADM were increased in duration by the adenosine 3',5'-cyclic monophosphate (cAMP) phosphodiesterase inhibitor rolipram but were not altered by inhibitors of guanosine 3',5'-cyclic monophosphate phosphodiesterase, nitric oxide synthetase, K(+)-ATP channels, the cyclooxygenase pathway, or the adrenergic nervous system. These results demonstrate that ADM-(22-52) is a selective CGRP receptor antagonist in the hindlimb vascular bed of the cat. The present data suggest that vasodilator responses to CGRP and ADM are mediated by different receptors but that these peptides dilate the hindlimb vascular bed of the cat by a similar cAMP-dependent mechanism.


PD-1 Inhibition Minimally Affects Cisplatin-Induced Toxicities in a Murine Model.
Immune checkpoint inhibition used in combination with standard cisplatin-based chemotherapy regimens is currently under evaluation in clinical trials for head and neck squamous cell carcinoma (HNSCC). The impact of anti-PD-1 therapy on cisplatin-induced ototoxicity and nephrotoxicity has not been established. Here we use a murine model of cisplatin-induced hearing loss to investigate the impact of anti-PD-1 immunotherapy on auditory brainstem responses (ABRs), distortion product otoacoustic emissions (DPOAEs), serum creatinine, and hair cell and renal histology. We demonstrate only mild worsening of DPOAEs at 14.4 and 16 kHz as well as a mild increase in serum creatinine. Renal and hair cell histology as well as ABR measures were unchanged by PD-1 inhibition. Thus, our data suggest that the use of PD-1 inhibition in conjunction with cisplatin results in toxicities that are similar to those of cisplatin alone.


Sam68 is absolutely required for Rev function and HIV-1 production.
Sam68 functionally complements for, as well as synergizes with, HIV-1 Rev in Rev response element (RRE)-mediated gene expression and virus production. Furthermore, C-terminal deletion/point mutants of Sam68 (Sam68DeltaC/Sam68-P21) exert a transdominant negative phenotype for Rev function and HIV-1 production. However, the relevance of Sam68 in Rev/RRE function is not well defined. To gain more insight into the mechanism of Sam68 in Rev function, we used an RNAi (RNA interference) strategy to create stable Sam68 knockdown HeLa (SSKH) cells. In SSKH cells, Rev failed to activate both RRE-mediated reporter gene [chloramphenicol acetyltransferase (CAT) and/or gag] expressions. Importantly, reduction of Sam68 expression led to a dramatic inhibition of HIV-1 production. Inhibition of the reporter gene expression and HIV production correlated with the failure to export RRE-containing CAT mRNA and unspliced viral mRNAs to the cytoplasm, confirming that SSKH cells are defective for Rev-mediated RNA export. Taken together, these results suggest that Sam68 is involved in Rev-mediated RNA export and is absolutely required for HIV production.


[Treatment of diabetes mellitus with long-acting biguanides].
The paper discusses of the results of treatment with preparations of phenylethylbiguanide (dibotin, meltrol, dipar, dibophen-retard), of butylbiguanide (silubin-retard, buforming-retard) and of dimethyl-biguanide (glucophage-retard). All these preparations were of prolonged action. The treatment was carried out in 242 patients. The saccharolytic action of the active agent contained in one tablet of each type of biguanide was approximately the same. Biguanides of prolonged action were highly effective in obese patients with diabetes mellitus of moderate severity. The best results were obtained in complex treatment with biguanides of prolonged action together with sulfonylurea preparations of the second generation. There were noted almost no toxic reactions from the use of biguanides up to 2 tablets a day.


Cisplatin chemotherapy for treatment of thyroid carcinoma in dogs: 13 cases.
The medical records of 13 dogs (10 spayed females, one intact male, one castrated male, and one dog of unknown gender) with histopathologically diagnosed thyroid carcinoma that were treated with cisplatin chemotherapy were reviewed. The mean age was 10.6 years. Three of the dogs were beagles. One dog had a c-cell medullary carcinoma, while the remaining 12 had follicular thyroid carcinoma. Eight of 13 dogs had tumors greater than 5 cm in diameter. In all 11 tumors for which information was available in the medical record, the masses were attached to underlying tissue. One dog underwent a complete remission, six had partial remissions, three had stable disease, and three had progressive disease. The mean time between initiation of cisplatin chemotherapy and development of progressive disease was 223.7 days (median, 202 days), with a mean survival time of 191.8 days (median, 98 days).


Apomorphine: modification of its hyperthermic effect in rabbits by p-chlorophenylalanine.
The hyperthermic response of rabbits to apomorphine, a dopaminergic agonist, is abolished by prior treatment with p-chlorophenylalanine. If such 5-hydroxytryptamine (5-HT)-depleted animals are administered a peripherally acting decarboxylase inhibitor plus 5-hydroxytryptophan, central stores of 5-HT are regenerated and the hyperthermic response to apomorphine is restored in part. The effects of apomorphine in rabbits with elevated concentrations of 5-HT are not different from those in control animals. The behavioral effects of apomnorphine appear to be constant in all groups of animals tested. It is suggested that the hyperthermic effects of apomorphine in rabbits require the presence of 5-HT.


Comparison of immunoadsorbents prepared by coupling sperm-whale myoglobin to a variety of insoluble polymers.
Immunoadsorbents that were potentially suitable for the fractionation of antibodies to myoglobin were prepared by coupling myoglobin to aminoethylcellulose, bromoacetylcellulose, CM-cellulose, an ethylene-maleic anhydride co-polymer and Sepharose 4B. A comparison of the properties of the preparations indicated that Sepharose-myoglobin was superior, as an immunoadsorbent, to the other conjugates. Elution of adsorbed antibodies with m-propionic acid, pH2.5, gave yields of precipitable antibody higher than those achieved with other dissociating solvents. Propionic acid eluted 60-100% of the antibody adsorbed by Sepharose-myoglobin, and 60-90% of the eluted protein was precipitable antibody.


Scaling Up Change: A Critical Review and Practical Guide to Harnessing Social Norms for Climate Action.
Anthropogenic carbon emissions have the potential to trigger changes in climate and ecosystems that would be catastrophic for the well-being of humans and other species. Widespread shifts in production and consumption patterns are urgently needed to address climate change. Although transnational agreements and national policy are necessary for a transition to a fully decarbonized global economy, fluctuating political priorities and lobbying by vested interests have slowed these efforts. Against this backdrop, bottom-up pressure from social movements and shifting social norms may offer a complementary path to a more sustainable economy. Furthermore, norm change may be an important component of decarbonization policies by accelerating or strengthening the impacts of other demand-side measures. Individual actions and policy support are social processes-they are intimately linked to expectations about the actions and beliefs of others. Although prevailing social norms often reinforce the status quo and unsustainable development pathways, social dynamics can also create widespread and rapid shifts in cultural values and practices, including increasing pressure on politicians to enact ambitious policy. We synthesize literature on social-norm influence, measurement, and change from the perspectives of psychology, anthropology, sociology, and economics. We discuss the opportunities and challenges for the use of social-norm and social-tipping interventions to promote climate action. Social-norm interventions aimed at addressing climate change or other social dilemmas are promising but no panacea. They require in-depth contextual knowledge, ethical consideration, and situation-specific tailoring and testing to understand whether they can be effectively implemented at scale. Our review aims to provide practitioners with insights and tools to reflect on the promises and pitfalls of such interventions in diverse contexts.


Function of heterologous Mycobacterium tuberculosis InhA, a type 2 fatty acid synthase enzyme involved in extending C20 fatty acids to C60-to-C90 mycolic acids, during de novo lipoic acid synthesis in Saccharomyces cerevisiae.
We describe the physiological function of heterologously expressed Mycobacterium tuberculosis InhA during de novo lipoic acid synthesis in yeast (Saccharomyces cerevisiae) mitochondria. InhA, representing 2-trans-enoyl-acyl carrier protein reductase and the target for the front-line antituberculous drug isoniazid, is involved in the activity of dissociative type 2 fatty acid synthase (FASII) that extends associative type 1 fatty acid synthase (FASI)-derived C(20) fatty acids to form C(60)-to-C(90) mycolic acids. Mycolic acids are major constituents of the protective layer around the pathogen that contribute to virulence and resistance to certain antimicrobials. Unlike FASI, FASII is thought to be incapable of de novo biosynthesis of fatty acids. Here, the genes for InhA (Rv1484) and four similar proteins (Rv0927c, Rv3485c, Rv3530c, and Rv3559c) were expressed in S. cerevisiae etr1Delta cells lacking mitochondrial 2-trans-enoyl-thioester reductase activity. The phenotype of the yeast mutants includes the inability to produce sufficient levels of lipoic acid, form mitochondrial cytochromes, respire, or grow on nonfermentable carbon sources. Yeast etr1Delta cells expressing mitochondrial InhA were able to respire, grow on glycerol, and produce lipoic acid. Commensurate with a role in mitochondrial de novo fatty acid biosynthesis, InhA could accept in vivo much shorter acyl-thioesters (C(4) to C(8)) than was previously thought (>C(12)). Moreover, InhA functioned in the absence of AcpM or protein-protein interactions with its native FASII partners KasA, KasB, FabD, and FabH. None of the four proteins similar to InhA complemented the yeast mutant phenotype. We discuss the implications of our findings with reference to lipoic acid synthesis in M. tuberculosis and the potential use of yeast FASII mutants for investigating the physiological function of drug-targeted pathogen enzymes involved in fatty acid biosynthesis.


Neural activity in frontal cortical cell layers: evidence for columnar sensorimotor processing.
The mammalian frontal cortex (FCx) is at the top of the brain's sensorimotor hierarchy and includes cells in the supragranular Layer 2/3, which integrate convergent sensory information for transmission to infragranular Layer 5 cells to formulate motor system outputs that control behavioral responses. Functional interaction between these two layers of FCx was examined using custom-designed ceramic-based microelectrode arrays (MEAs) that allowed simultaneous recording of firing patterns of FCx neurons in Layer 2/3 and Layer 5 in nonhuman primates performing a simple go/no-go discrimination task. This unique recording arrangement showed differential encoding of task-related sensory events by cells in each layer with Layer 2/3 cells exhibiting larger firing peaks during presentation of go target and no-go target task images, whereas Layer 5 cells showed more activity during reward contingent motor responses in the task. Firing specificity to task-related events was further demonstrated by synchronized firing between pairs of cells in different layers that occupied the same vertically oriented "column" on the MEA. Pairs of cells in different layers recorded at adjacent "noncolumnar" orientations on the MEA did not show synchronized firing during the same task-related events. The results provide required evidence in support of previously suggested task-related sensorimotor processing in the FCx via functionally segregated minicolumns.


Exosomal transfer of miR-126 promotes the anti-tumour response in malignant mesothelioma: Role of miR-126 in cancer-stroma communication.
MiR-126 has been shown to suppress malignant mesothelioma (MM) by targeting cancer-related genes without inducing toxicity or histopathological changes. Exosomes provide the opportunity to deliver therapeutic cargo to cancer stroma. Here, a tumour stromal model composed of endothelial cells (HUVECs), fibroblasts (IMR-90 cells), non-malignant mesothelial cells (Met-5A cells) and MM cells (H28 and MM-B1 cells) was used. The cells were treated with exosomes from HUVECs carrying endogenous (exo-HUVEC) and enriched miR-126 (exo-HUVECmiR-126), and the uptake/turnover of exosomes; miR-126 distribution within the stroma; and effect of miR-126 on cell signalling, angiogenesis and cell proliferation were evaluated. Based on the sensitivity of MM cells to exo-HUVEC miR-126 treatment, miR-126 was distributed differently across stromal cells. The reduced miR-126 content in fibroblasts in favour of endothelial cells reduced angiogenesis and suppressed cell growth in an miR-126-sensitive environment. Conversely, the accumulation of miR-126 in fibroblasts and the reduced level of miR-126 in endothelial cells induced tube formation in an miR-126-resistant environment via VEGF/EGFL7 upregulation and IRS1-mediated cell proliferation. These findings suggest that transfer of miR-126 via HUVEC-derived exosomes represents a novel strategy to inhibit angiogenesis and cell growth in MM.


Adjuvant and Salvage Radiotherapy after Prostatectomy: ASTRO/AUA Guideline Amendment 2018-2019.
The purpose of this amendment is to incorporate newly-published literature into the original ASTRO/AUA Adjuvant and Salvage Radiotherapy after Prostatectomy Guideline and to provide an updated clinical framework for clinicians. The original systematic review yielded 294 studies published between January 1990 and December 2012. In April 2018, the guideline underwent an amendment and incorporated 155 references that were published from January 1990 through December 2017. Two new key questions were added. One on the use of genomic classifiers and the other on the treatment of oligo-metastases with radiation post-radical prostatectomy. A new statement on the use of hormone therapy with salvage radiotherapy after radical prostatectomy was added and long-term data was used to update an existing statement on adjuvant radiotherapy. The balance of the guideline statements were re-affirmed and references were added to the existing literature base. A discussion on the use of genomic classifiers as a risk stratification tool was added to the future research discussion. No relevant data on oligo-metastases was found. Hormone therapy should be offered to patients who have had radical prostatectomy and who are candidates for salvage radiotherapy. The clinician should discuss possible short- and long-term side effects with the patient as well as the potential benefits of preventing recurrence. The decision to use hormone therapy should be made by the patient and a multi-disciplinary team of providers with full consideration of the patient's history, values, preferences, quality of life, and functional status.


Identification and prioritisation of risks in a hospital pharmacy using healthcare failure mode and effect analysis.
The goals of this project included identifying the processes and subprocesses performed in hospital pharmacies, identifying potential adverse events, detecting failure modes and the causes of errors, prioritising the risks identified and designing a map of risks for hospital pharmacies. A task force composed of hospital pharmacy staff was committed to update the diagram of processes and design a map of processes performed in hospital pharmacies. Risks were identified by failure mode and effect analysis annd prioritised according to their risk priority index (RPI) and criticality. A risk map of adverse events was designed based on the diagram of processes and/or primary activities where the prioritised failure modes were most frequent. In total, 99 failure modes associated with 80 adverse events and 129 causes were identified in eight hospital pharmacy areas/subprocesses. The three areas with the highest percentages of failure modes were inpatient pharmaceutical care, pharmacy laboratory and pharmaceutical technology, and medication management. The 25 failure modes (first quartile) with the highest RPI scores (RPI≥20) and the 25 failure modes with the highest frequency and criticality scores were classified as priority. According to their RPI, priority failure modes mostly occurred in the area of inpatient pharmaceutical care (92%). However, according to their criticality, priority failure modes were found to homogeneously occur across all pharmaceutical care areas. As general recommendations pharmacists should assume responsibility and leadership in the implementation of safe medication use practices in healthcare centres.


One-Pot Green Synthesis of Biocompatible Graphene Quantum Dots and Their Cell Uptake Studies.
Graphene-based quantum dots (GQDs) are attractive fluorophores due to their excellent photoluminescence properties, water solubility, low cost, and low toxicity. However, the lack of simple, efficient, and environmental-friendly synthesis methods often limits their biological applications. Herein, we explore a novel, one-pot, green synthesis approach for the fabrication of fluorescent GQDs without involving any harsh reagents. Graphene oxide is used as a precursor, and a 2 h hydrothermal synthesis is carried out with assistance of hydrogen peroxide; no further post purification steps are required. The effects of reaction conditions on the characteristics of GQDs are comprehensively investigated. The as-synthesized GQDs show a high photostability and excellent biocompatibility as revealed by cell viability assays for three different cell lines, namely, macrophages, endothelial cells, and a model cancer cell line. The detailed studies of cellular uptake mechanisms suggest that for all of the three cell lines the major internalization route for GQDs is caveolae-mediated endocytosis followed by clathrin-mediated endocytosis at a less extent. Our results demonstrate the great potential of the as-synthesized GQDs as fluorescent nanoprobes. The study also provides unique insight into the cell-GQDs interactions, which is highly valuable for bioimaging and other related applications such as diagnostics and drug delivery.


The electron-phonon scattering and carrier mobility in monolayer AsSb.
Through first-principles calculations, the electron-phonon scattering of two-dimensional monolayer AsSb is investigated. The Boltzmann transport equation is used to compute the phonon limited carrier mobility. We find that the optical phonon scattering rates are much larger than acoustic ones around the valence band maximum (VBM) and the conduction band minimum (CBM). The phonon-decomposed scattering results show that transverse optical (TO) phonon modes dominate the scattering around VBM, while longitudinal and out-of-plane acoustic modes contribute mostly to the scattering at higher energy. TO phonon modes dominate the scattering for electrons over all energy level, and the electron-phonon matrix element analysis verified the results. Finally, we observed that the largest mean free paths for hot holes and hot electrons are 20 nm and 8 nm, respectively. That is the best range to extract the hot holes and hot electrons. More interestingly, owing to the in-pane net dipole moment of AsSb, the intrinsic electron/hole mobility of AsSb are 38/50 cm2 V-1 s-1, which are less than monolayer arsenene and antimonene.


Phenotypic consequences of red-white colony type variation in Mycobacterium avium.
Mycobacterium avium undergoes reversible morphotypic switching between the virulent transparent colony type and the less virulent opaque colony type. A new morphotypic switch in M. avium, termed red-white, that becomes visible when opaque colonies of clinical isolates are grown on agar media containing Congo red, was recently described. White opaque (WO) variants were found to be more resistant to multiple antibiotics than were red opaque (RO) variants. The present paper reports that transparent derivatives of RO and WO clones retain the differential Congo red binding properties of their opaque parents, indicating that the opaque-transparent switch operates independently of the red-white switch. White transparent variants were more resistant to clarithromycin and rifampin in vitro, and better able to survive within human macrophages, than their red transparent counterparts. Neither red nor white variants were markedly favoured during growth in vitro; however, red variants were better able to spread on soft agar (sliding motility), a potential selective advantage under some environmental circumstances. White-to-red switching was frequently observed in vitro and was accompanied by decreased antibiotic resistance and increased motility. Red-to-white switching has yet to be observed in vitro, indicating that the red morphotype is very stable. Significantly, some widely studied laboratory reference strains of M. avium, including strain 2151 and the genome sequence strain 104, are stable red clones. These strains are intrinsically antibiotic resistant and virulent in animal models, but they may not express genes encoding the elevated levels of antibiotic resistance and intracellular survival observed in white variants.


Hydroxychloroquine decreases human MSC-derived osteoblast differentiation and mineralization in vitro.
We recently showed that patients with primary Sjögren Syndrome (pSS) have significantly higher bone mineral density (BMD) compared to healthy controls. The majority of those patients (69%) was using hydroxychloroquine (HCQ), which may have favourable effects on BMD. To study the direct effects of HCQ on human MSC-derived osteoblast activity. Osteoblasts were cultured from human mesenchymal stromal cells (hMSCs). Cultures were treated with different HCQ doses (control, 1 and 5 µg/ml). Alkaline phosphatase activity and calcium measurements were performed to evaluate osteoblast differentiation and activity, respectively. Detailed microarray analysis was performed in 5 µg/ml HCQ-treated cells and controls followed by qPCR validation. Additional cultures were performed using the cholesterol synthesis inhibitor simvastatin (SIM) to evaluate a potential mechanism of action. We showed that HCQ inhibits both MSC-derived osteoblast differentiation and mineralization in vitro. Microarray analysis and additional PCR validation revealed a highly significant up-regulation of the cholesterol biosynthesis, lysosomal and extracellular matrix pathways in the 5 µg/ml HCQ-treated cells compared to controls. Besides, we demonstrated that 1 µM SIM also decreases MSC-derived osteoblast differentiation and mineralization compared to controls. It appears that the positive effect of HCQ on BMD cannot be explained by a stimulating effect on the MSC-derived osteoblast. The discrepancy between high BMD and decreased MSC-derived osteoblast function due to HCQ treatment might be caused by systemic factors that stimulate bone formation and/or local factors that reduce bone resorption, which is lacking in cell cultures.


Hemodynamic improvement in cardiac resynchronization does not require improvement in left ventricular rotation mechanics: three-dimensional tagged MRI analysis.
Earlier studies have yielded conflicting evidence on whether or not cardiac resynchronization therapy (CRT) improves left ventricular (LV) rotation mechanics. In dogs with left bundle branch block and pacing-induced heart failure (n=7), we studied the effects of CRT on LV rotation mechanics in vivo by 3-dimensional tagged magnetic resonance imaging with a temporal resolution of 14 ms. CRT significantly improved hemodynamic parameters but did not significantly change the LV rotation or rotation rate. LV torsion, defined as LV rotation of each slice with respect to that of the most basal slice, was not significantly changed by CRT. CRT did not significantly change the LV torsion rate. There was no significant circumferential regional heterogeneity (anterior, lateral, inferior, and septal) in LV rotation mechanics in either left bundle branch block with pacing-induced heart failure or CRT, but there was significant apex-to-base regional heterogeneity. CRT acutely improves hemodynamic parameters without improving LV rotation mechanics. There is no significant circumferential regional heterogeneity of LV rotation mechanics in the mechanically dyssynchronous heart. These results suggest that LV rotation mechanics is an index of global LV function, which requires coordination of all regions of the left ventricle, and improvement in LV rotation mechanics appears to be a specific but insensitive index of acute hemodynamic response to CRT.


Uptake and transport of herpes simplex virus in neurites of rat dorsal root ganglia cells in culture.
Attachment and neuritic transport of herpes simplex virus (HSV) type 1 (McIntyre) were studied in a cell culture system with dissociated cells of rat dorsal root ganglia. The two-chamber cell culture system containing a diffusion barrier penetrated by neurites of cultured sensory neurons permitted infection of neurites extending outside the diffusion barrier without exposure of the neuronal cell soma. HSV adsorbed to neuritic extensions and reached the neuronal soma within 1.5 h post-inoculation. Neuritic uptake and transport of HSV were inhibited in the presence of cytochalasin B. Internalization of virus in neurites was preceded by attachment of virus to the neurite plasma membrane. Neurites transported viral nucleocapsids (NC) through the diffusion barrier of the cultures. Destruction of the neuritic extensions before or shortly after peripheral virus inoculation blocked spread of infection to the cell soma. No infection was established when neuritic extensions were exposed to viral NC and NC were then not observed inside the neurite plasma membrane. Virus produced in neurons, when HSV was inoculated into the inner culture chamber containing the neuronal cell bodies, was transported as enveloped virus in cytoplasmic vesicles from the neuronal cell body towards the periphery. Schwann cells were infected by viropexis. Shortly after infection virions were observed in vacuoles of the cytoplasm.


How does human motor cortex regulate vocal pitch in singers?
Vocal pitch is used as an important communicative device by humans, as found in the melodic dimension of both speech and song. Vocal pitch is determined by the degree of tension in the vocal folds of the larynx, which itself is influenced by complex and nonlinear interactions among the laryngeal muscles. The relationship between these muscles and vocal pitch has been described by a mathematical model in the form of a set of 'control rules'. We searched for the biological implementation of these control rules in the larynx motor cortex of the human brain. We scanned choral singers with functional magnetic resonance imaging as they produced discrete pitches at four different levels across their vocal range. While the locations of the larynx motor activations varied across singers, the activation peaks for the four pitch levels were highly consistent within each individual singer. This result was corroborated using multi-voxel pattern analysis, which demonstrated an absence of patterned activations differentiating any pairing of pitch levels. The complex and nonlinear relationships between the multiple laryngeal muscles that control vocal pitch may obscure the neural encoding of vocal pitch in the brain.


Diastereoselective synthesis of potent antimalarial cis-β-lactam agents through a [2 + 2] cycloaddition of chiral imines with a chiral ketene.
The effect of double asymmetric induction for the synthesis of new cis-β-lactams by [2 + 2] cycloaddition reactions of chiral imines with a chiral ketene was investigated. The cycloaddition reaction was found to be totally diastereoselective leading exclusively to the formation of the cis-β-lactam derivatives. The newly synthesized cycloadducts were evaluated for their antimalarial activities against Plasmodium falciparum K14 resistant strain with moderate to excellent IC50 values varying from 8 to 50 μM. Of the fifteen β-lactams tested, four showed IC50 ≤ 11 μM.


Relationship between rice farming and polygenic scores potentially linked to agriculture in China.
Following domestication in the lower Yangtze River valley 9400 years ago, rice farming spread throughout China and changed lifestyle patterns among Neolithic populations. Here, we report evidence that the advent of rice domestication and cultivation may have shaped humans not only culturally but also genetically. Leveraging recent findings from molecular genetics, we construct a number of polygenic scores (PGSs) of behavioural traits and examine their associations with rice cultivation based on a sample of 4101 individuals recently collected from mainland China. A total of nine polygenic traits and genotypes are investigated in this study, including PGSs of height, body mass index, depression, time discounting, reproduction, educational attainment, risk preference, ADH1B rs1229984 and ALDH2 rs671. Two-stage least-squares estimates of the county-level percentage of cultivated land devoted to paddy rice on the PGS of age at first birth (b = -0.029, p = 0.021) and ALDH2 rs671 (b = 0.182, p < 0.001) are both statistically significant and robust to a wide range of potential confounds and alternative explanations. These findings imply that rice farming may influence human evolution in relatively recent human history.


The characterization of atrial natriuretic peptide (ANP) expression by primary cultures of atrial myocytes using an ANP-specific monoclonal antibody and an ANP messenger ribonucleic acid probe.
The biochemical and morphological characteristics of primary neonatal rat atrial myocytes were examined in order to establish a model system for future studies of the biosynthesis and secretion of atrial natriuretic peptide (ANP). Preliminary studies demonstrated that the quantity of immunoactive ANP/microgram protein within rat atria increased as a function of age from 2 ng/micrograms in 19 day prenatal animals to 400 ng/micrograms in the adult. Gel filtration, reversed phase HPLC, and ion exchange HPLC indicated that there were similar quantities of immunoactive ANP in the right and left atria at various ages, and that the major molecular form of the peptide in the heart is chromatographically indistinguishable from ANP(1-126). Cultures of dissociated cells were prepared from pooled left and right atria derived from 1 day postnatal animals. A complete serum-free medium was developed which resulted in the maintenance of high levels of immunoactive ANP in the cultures. As determined by RIA, the cellular content of ANP increased in the cultures as a function of time through 7 days in vitro. The quantity of immunoactive ANP in the cultures increased approximately 2- to 3-fold between days 3 and 7. When the cultures that had been maintained for 7 days were submitted to immunocytochemistry using an ANP-specific monoclonal antibody, distinct colonies of spindle-shaped cells stained positively. In situ hybridization, utilizing an 35S-labeled ANP messenger RNA probe, demonstrated that these colonies of myocytes expressed the ANP message. Using quantitative dot-blot hybridization it was shown that the ANP mRNA level increased approximately 50-fold between days 1 and 7 in culture. These studies indicate that the serum-free culture medium allows continued accumulation of both ANP and the ANP message in culture and will provide a useful model system to characterize factors that regulate the biosynthesis and secretion of this hormone.


In situ chlorpyrifos (CPF) degradation by Acrobeloides maximus: Insights from chromatographic analysis.
The objective of this study was to evaluate the efficiency of nematodes in zooremediation of chlorpyrifos (CPF), an organophosphate pesticide. The nematode population Acrobeloides maximus (A. maximus) was employed for bioremediation, converting CPF into non-toxic residues. Optimal growth conditions for mass production of A. maximus were achieved by maintaining a temperature of 25 °C, pH 8, and supplementing the culture medium with plant nutrients. The nematodes were then immobilized within sodium alginate beads. The efficacy of the degradation process was assessed using various analytical techniques, including UV-Visible spectroscopy, HPTLC, FTIR, and LC-MS, confirming the successful breakdown of CPF. The bioreactor demonstrated a complete degradation efficiency of CPF exceeding 99%. Additionally, LC-MS analysis was conducted to elucidate the degradation pathway based on the formation of intermediates. These results underscore the potential of A. maximus as a sustainable organism for addressing environmental contamination arising from CPF pesticide.


Novel Cr/Si-Slurry Diffusion Coatings for High Temperatures.
Surface enrichment in Al, Si, and Cr can greatly improve high temperature oxidation resistance of many alloys. Al, Si, and Cr coatings are commonly applied via simple slurries or more complex pack cementation processes. Due to the high melting point of Cr, the deposition of Cr-based diffusion coatings by the slurry technique has proved challenging, and to date, Cr has mostly been applied by pack cementation. Here, a novel Cr-Si coating process via the slurry technique is described which has been developed and then demonstrated on two Ni-based superalloys, Rene 80 and Inconel 740H. The addition of Si to the slurry lowers the melting point via a Cr-Si eutectic and enables the formation of a liquid phase during heat treatment. Through this Cr-Si slurry coating process diffusion layers enriched by Cr and Si of about 150 µm were achieved. Oxidation behavior was studied through isothermal exposures at 900 °C for 1000 h in lab air. Uncoated Rene 80 and IN740H both showed formation of a Ti-containing Cr2O3 scale below a thin TiO2 top layer. Underneath the external scale a zone of internally oxidized Al grew over the exposure time and reduced the load-bearing cross-section progressively. In comparison, the Cr/Si-coated samples did not show internal Al oxidation, but a slow-growing Si-rich oxide film underneath the external Cr2O3 scale. This subscale represents an additional oxygen diffusion barrier. Thus, the weight gain during exposure for the coated samples was significantly lower than for the uncoated materials.


Discordance of skin and muscle involvement in dermatomyositis.
The skin manifestations of dermatomyositis infrequently occur without the myositis. A 24-year-old woman presented with concordance of the skin and muscle components of dermatomyositis, followed by a remission of the myositis with a persistence of significant rash for 17 years, finally presenting with a flare of both skin and muscle components together. Clinicians should be alert to the recurrence of an underlying myositis at any time.


From tissue to silicon to plastic: three-dimensional printing in comparative anatomy and physiology.
Comparative anatomy and physiology are disciplines related to structures and mechanisms in three-dimensional (3D) space. For the past centuries, scientific reports in these fields have relied on written descriptions and two-dimensional (2D) illustrations, but in recent years 3D virtual modelling has entered the scene. However, comprehending complex anatomical structures is hampered by reproduction on flat inherently 2D screens. One way to circumvent this problem is in the production of 3D-printed scale models. We have applied computed tomography and magnetic resonance imaging to produce digital models of animal anatomy well suited to be printed on low-cost 3D printers. In this communication, we report how to apply such technology in comparative anatomy and physiology to aid discovery, description, comprehension and communication, and we seek to inspire fellow researchers in these fields to embrace this emerging technology.


Biphasic Release of the Alarmin High Mobility Group Box 1 Protein Early After Trauma Predicts Poor Clinical Outcome.
The causal role of the prototype alarmin high mobility group box 1 protein in systemic inflammation and remote organ injury after trauma and shock is established in animal models but not in humans. Our aim was therefore to determine high mobility group box 1 protein concentration kinetics with high time resolution during the first hours after trauma in individual patients and investigate the association with outcome. Prospective single-center observational study. University hospital Level I trauma center. Convenience recruitment of 136 trauma patients. None. Total plasma high mobility group box 1 protein levels were analyzed with enzyme-linked immunosorbent assay in repeated samples. Relationships between predefined predictor variables and outcome were examined in multivariable linear regression models. Ventilator-free days was used as primary outcome measure. Two distinct high mobility group box 1 protein release phases were identified. An initial exponential decay phase with half-life 26 minutes was not correlated with outcome. In contrast, a second high mobility group box 1 protein wave peaking 3-6 hours after trauma in the most severely injured and physiologically deranged patients was consistently the most important predictor of outcome in our multivariable models, rendering all other predictor variables insignificant except for smaller contributions from age and sex, and of admission base excess for maximal creatinine concentration. High mobility group box 1 protein was released in two consecutive phases. Only the second high mobility group box 1 protein wave was a significant predictor of outcome. Patients with a high high mobility group box 1 protein concentration between 3 and 6 hours after trauma might hypothetically benefit from high mobility group box 1 protein-specific antagonist therapy.


Phosphorus losses through agricultural tile drainage in Nova Scotia, Canada.
Tile drainage water from agricultural fields commonly exceeds environmental guidelines for phosphorus (P) in rivers and streams. The loss of P through artificial drainage is spatially and temporally variable, and is related to local factors. This study characterizes variability in total P (TP) and soluble reactive P (SRP) concentrations in weekly drainage samples from 39 agricultural fields in Nova Scotia, Canada, from April 2002 through December 2003. We examined connections between P concentrations and the factors: (i) soil texture; (ii) discharge flow rate; (iii) soil test P (STP); (iv) manure type; and (v) crop cover. Generally, variability between fields and samples was great, and fields with standard deviations exceeding the mean for TP, SRP, and flow rate were 71, 54, and 79%, respectively. It was evident that poultry and swine manure contributed to high STPs, and to constantly high TP concentrations with high proportions of SRP. Concentrations varied from week to week, and particularly in April, May, October, and November when the greatest TP, SRP, and flow rate averages were measured. Mean TP concentrations exceed the USEPA (1994) TP guideline of 0.10 mg L(-1) at 82% of the fields, and periodically concentrations more than 10 times, and occasionally more than 50 times higher than the guideline were found. The proportion of SRP in TP had a tendency to be higher when TP levels were high in coarse textured soils. In Nova Scotia, dairy manure is most often applied on permanent cover crops, which did not show as much P concentration variability as crop rotations. Daily or hourly observation of short-term increases in P concentrations related to the described factors would help to characterize the changes in P concentrations observed during frequent heavy drainage flow events.


S. Typhimurium sseJ gene decreases the S. Typhi cytotoxicity toward cultured epithelial cells.
Salmonella enterica serovar Typhi and Typhimurium are closely related serovars as indicated by >96% DNA sequence identity between shared genes. Nevertheless, S. Typhi is a strictly human-specific pathogen causing a systemic disease, typhoid fever. In contrast, S. Typhimurium is a broad host range pathogen causing only a self-limited gastroenteritis in immunocompetent humans. We hypothesize that these differences have arisen because some genes are unique to each serovar either gained by horizontal gene transfer or by the loss of gene activity due to mutation, such as pseudogenes. S. Typhi has 5% of genes as pseudogenes, much more than S. Typhimurium which contains 1%. As a consequence, S. Typhi lacks several protein effectors implicated in invasion, proliferation and/or translocation by the type III secretion system that are fully functional proteins in S. Typhimurium. SseJ, one of these effectors, corresponds to an acyltransferase/lipase that participates in SCV biogenesis in human epithelial cell lines and is needed for full virulence of S. Typhimurium. In S. Typhi, sseJ is a pseudogene. Therefore, we suggest that sseJ inactivation in S. Typhi has an important role in the development of the systemic infection. We investigated whether the S. Typhi trans-complemented with the functional sseJ gene from S. Typhimurium (STM) affects the cytotoxicity toward cultured cell lines. It was found that S. Typhi harbouring sseJSTM presents a similar cytotoxicity level and intracellular retention/proliferation of cultured epithelial cells (HT-29 or HEp-2) as wild type S. Typhimurium. These phenotypes are significantly different from wild type S. Typhi Based on our results we conclude that the mutation that inactivate the sseJ gene in S. Typhi resulted in evident changes in the behaviour of bacteria in contact with eukaryotic cells, plausibly contributing to the S. Typhi adaptation to the systemic infection in humans.


Biological treatability of raw and ozonated penicillin formulation effluent.
In the present study, oxidative pre-treatment of pharmaceutical wastewater originating from the formulation of the penicillin Sultamycillin Tosylate Diydrate via ozonation at varying pH and ozone feed rates was investigated. Biological treatability studies were performed with a synthetic wastewater alone and supplemented with raw and ozonated penicillin formulation effluents. The highest COD (34%) and TOC (24%) removal efficiencies were obtained at pH 11.0, whereas the BOD5 value increased from 16 mg l(-1) to 128 mg l(-1) after 40 min of ozonation, corresponding to an applied ozone dose of 1670 mg l(-1) and 33% relative ozone absorption. The studies showed that no degradation of raw penicillin fraction (30% of total COD) occurred, and degradation of the synthetic wastewater being completely treatable without penicillin addition, was inhibited by 7%. Upon 40 min ozonation, the synthetic wastewater could be completely oxidized and at the same time 35% of ozonated penicillin wastewater removal was obtained. Respirometric studies were conducted in parallel and produced results indicating a 22% decrease in the total oxygen consumption rate established for raw penicillin formulation effluent compared to the results obtained from the aerobic batch reactor. No inhibition of the synthetic fraction was observed for the 40 min-ozonated penicillin formulation effluent, biodegradability of the 60 min-ozonated penicillin effluent decreased possibly due to recalcitrant oxidation product accumulation. The modeling study provided experimental support and information on inhibition kinetics in activated sludge model no. 3 (ASM3) by means of respirometric tests for the first time.


Effects of nuclear cross sections at different energies on the radiation hazard from galactic cosmic rays.
The radiation hazard for astronauts from galactic cosmic rays (GCR) is a major obstacle to long-duration human space exploration. Space radiation transport codes have been developed to calculate the radiation environment on missions to the Moon, Mars, and beyond. We have studied how uncertainties in fragmentation cross sections at different energies affect the accuracy of predictions from such radiation transport calculations. We find that, in deep space, cross sections at energies between 0.3 and 0.85 GeV/nucleon have the largest effect in solar maximum GCR environments. At the International Space Station, cross sections at higher energies have the largest effect due to the geomagnetic cutoff.


Orthogonal search-based rule extraction (OSRE) for trained neural networks: a practical and efficient approach.
There is much interest in rule extraction from neural networks and a plethora of different methods have been proposed for this purpose. We discuss the merits of pedagogical and decompositional approaches to rule extraction from trained neural networks, and show that some currently used methods for binary data comply with a theoretical formalism for extraction of Boolean rules from continuously valued logic. This formalism is extended into a generic methodology for rule extraction from smooth decision surfaces fitted to discrete or quantized continuous variables independently of the analytical structure of the underlying model, and in a manner that is efficient even for high input dimensions. This methodology is then tested with Monks' data, for which exact rules are obtained and to Wisconsin's breast cancer data, where a small number of high-order rules are identified whose discriminatory performance can be directly visualized.


Telephone-administered versus live group cognitive behavioral stress management for adults with CFS.
Chronic fatigue syndrome (CFS) symptoms have been shown to be exacerbated by stress and ameliorated by group-based psychosocial interventions such as cognitive behavioral stress management (CBSM). Still, patients may have difficulty attending face-to-face groups. This study compared the effects of a telephone-delivered (T-CBSM) vs a live (L-CBSM) group on perceived stress and symptomology in adults with CFS. Intervention data from 100 patients with CFS (mean age 50years; 90% female) participating in T-CBSM (N=56) or L-CBSM (N=44) in previously conducted randomized clinical trials were obtained. Perceived Stress Scale (PSS) and the Centers for Disease Control and Prevention symptom checklist scores were compared with repeated measures analyses of variance in adjusted and unadjusted analyses. Participants across groups showed no differences in most demographic and illness variables at study entry and had similar session attendance. Both conditions showed significant reductions in PSS scores, with L-CBSM showing a large effect (partial ε2=0.16) and T-CBSM a medium effect (partial ε2=0.095). For CFS symptom frequency and severity scores, L-CBSM reported large effect size improvements (partial ε2=0.19-0.23), while T-CBSM showed no significant changes over time. Two different formats for delivering group-based CBSM-live and telephone-showed reductions in perceived stress among patients with CFS. However, only the live format was associated with physical symptom improvements, with specific effects on post-exertional malaise, chills, fever, and restful sleep. The added value of the live group format is discussed, along with implications for future technology-facilitated group interventions in this population.


On-chip actuation transmitter for enhancing the dynamic response of cell manipulation using a macro-scale pump.
An on-chip actuation transmitter for achieving fast and accurate cell manipulation is proposed. Instead of manipulating cell position by a directly connected macro-scale pump, polydimethylsiloxane deformation is used as a medium to transmit the actuation generated from the pump to control the cell position. This actuation transmitter has three main advantages. First, the dynamic response of cell manipulation is faster than the conventional method with direct flow control based on both the theoretical modeling and experimental results. The cell can be manipulated in a simple harmonic motion up to 130 Hz by the proposed actuation transmitter as opposed to 90 Hz by direct flow control. Second, there is no need to fill the syringe pump with the sample solution because the actuation transmitter physically separates the fluids between the pump and the cell flow, and consequently, only a very small quantity of the sample is required (<1 μl). In addition, such fluid separation makes it easy to keep the experiment platform sterilized because there is no direct fluid exchange between the sample and fluid inside the pump. Third, the fabrication process is simple because of the single-layer design, making it convenient to implement the actuation transmitter in different microfluidic applications. The proposed actuation transmitter is implemented in a lab-on-a-chip system for red blood cell (RBC) evaluation, where the extensibility of red blood cells is evaluated by manipulating the cells through a constriction channel at a constant velocity. The application shows a successful example of implementing the proposed transmitter.


Promoter-specific alterations of APC are a rare cause for mutation-negative familial adenomatous polyposis.
n familial adenomatous polyposis (FAP), 20% of classical and 70% of attenuated/atypical (AFAP) cases remain mutation-negative after routine testing; yet, allelic expression imbalance may suggest an APC alteration. Our aim was to determine the proportion of families attributable to genetic or epigenetic changes in the APC promoter region. We studied 51 unrelated families/cases (26 with classical FAP and 25 with AFAP) with no point mutations in the exons and exon/intron borders and no rearrangements by multiplex ligation-dependent probe amplification (MLPA, P043-B1). Promoter-specific events of APC were addressed by targeted resequencing, MLPA (P043-C1), methylation-specific MLPA, and Sanger sequencing of promoter regions. A novel 132-kb deletion encompassing the APC promoter 1B and upstream sequence occurred in a classical FAP family with allele-specific APC expression. No promoter-specific point mutations or hypermethylation were present in any family. In conclusion, promoter-specific alterations are a rare cause for mutation-negative FAP (1/51, 2%). The frequency and clinical correlations of promoter 1B deletions are poorly defined. This investigation provides frequencies of 1/26 (4%) for classical FAP, 0/25 (0%) for AFAP, and 1/7 (14%) for families with allele-specific expression of APC. Clinically, promoter 1B deletions may associate with classical FAP without extracolonic manifestations.


Effect of milk flow-rate switch-point settings on milking duration and udder health throughout lactation.
The objective of this study was to quantify the effects of different milk flow-rate switch-point settings on milking duration, SCC, strip milk, teat condition and milk yield in a grass based system in a long-term experiment. Much work has already been conducted providing strong support for significant reduction in milking duration without impact on yield through increasing the flow-rate switch-point at which vacuum to the milking cluster ceases and the cluster is removed from the cow by means of a retracting cord. However, in practice many farms have not adopted this labor saving technology on the basis that it may increase milk SCC. Recent research on commercial Irish dairy herds identified the contagious mastitis causing pathogen Staph. aureus as the most prevalent pathogen detected. Staph. aureus could have a cyclical shedding pattern which would inhibit detection at certain time points. Therefore, to reliably assess the effect of milk flow-rate switch-points on SCC, a long-term study was required, consisting of multiple observations on cow level SCC. The present study filled this gap in knowledge by informing on any impact that ceasing milking at different flow-rates may have on milking duration and SCC levels, particularly with regard to spring calving grass based systems. Four treatments, consisting of milk flow-rate switch-points increasing from 0.2 kg/min to 0.8 kg/min in steps of 0.2 kg/min, were deployed for 31 weeks to cows at the Teagasc Research Centre at Moorepark, Ireland. The effect of treatment on daily milking duration was significant (P < 0.001). The milking duration for a milk flow-rate switch-point of 0.8 kg/min was 95 s (14%) shorter than for 0.2 kg/min. We did not find a significant effect of increasing the milk flow-rate switch-point from 0.2 to 0.8 kg/min on milk yield or SCC in this long-term study. We did find a significant effect of week of experiment on milk SCC, whereby the SCC of the cows on the experiment increased similarly among treatment groups as lactation progressed. A significant reduction in Dead Time (time from cluster attachment to reach a milk flow-rate of 0.2 kg/min) was also noted as the milk flow-rate switch-point increased. On average reductions in Dead Time contributed 12% to the overall reductions in Milking Duration. Similarly, reductions in Low Flow Time (time from a flow-rate of 0.2 kg/min to cluster detachment at the end of milking) contributed 26% to the overall reductions in Milking Duration. Reductions in Dead Time and Low Flow Time played a greater role in reducing PM Milking Duration rather than AM Milking Duration due to the milking interval practiced on the research farm.


Gonadotropin-induced proliferation of endoplasmic reticulum in an androgenic tumor and its relation to elevated plasma testosterone levels.
The development of gonadotropin-induced structural change in a testosterone-secreting Leydig cell tumor and its relationship to plasma testosterone levels were studied in castrate, tumor-bearing mice 2 hr after daily injections of human chorionic gonadotropin (HCG). Tumor cells from control animals were small, averaging less than 1.0 X 10(-9) ml in volume, and were poorly differentiated, having very little smooth endoplasmic reticulum (SER) in their cytoplasm. Average plasma testosterone levels in these mice were near 1.3 ng/ml. Although a five-fold rise in plasma testosterone was measured 2 hr after the first HCG injection, no changes in the endoplasmic reticulum were detected at this time. After the second injection, plasma testosterone rose to only twice control levels, while ultra-areas of mixed rough endoplasmic reticulum and SER. Modest amounts of SER had appeared in many tumor cells after the third injection, and plasma testosterone showed a six-fold rise. Between the third and fourth injections, cell volume increased by about 70% as large accumulations of SER appeared in the cytoplasm of most cells. Plasma testosterone again rose to five times control levels. Increased cell volume and abundant SER were maintained by continuing daily injections of HCG, while the 2-hr plasma testosterone response persisted at five to six times the control level. These findings show that a maximal elevation of plasma testosterone can occur prior to changes in the endoplasmic reticulum of Leydig tumor cells. However, they also suggest that daily repetition of the maximal functional response requires that the cells acquire large quantities of newly produced SER.


Effect of single-dose ivermectin therapy on human Onchocerca volvulus infection with onchocercal ocular involvement.
Ivermectin has shown promise as a potentially safe and effective microfilaricidal drug for the treatment of onchocerciasis. Several limited studies have shown it to have fewer side effects, especially ocular complications, than the currently available drug, diethylcarbamazine. The detailed ocular findings in 200 moderately to heavily infected Liberians who were enrolled in a safety and dose-finding study are presented. They received either 0, 100, 150, or 200 micrograms/kg of ivermectin and were followed up for 12 months. In clinical studies so far carried out ivermectin in a dose of 100, 150, or 200 micrograms/kg has not been associated with any major adverse reactions nor were there any sight-threatening effects even in the presence of severe ocular disease. Each of these doses significantly reduced the ocular microfilaria load for at least 12 months when compared with either the placebo (p less than 0.05) or pretreatment values (p less than 0.001). However, the 100 and 150 micrograms/kg doses caused fewer minor side effects than the higher dose. These results confirm that ivermectin in a single oral dose may be a safe and effective microfilaricidal drug for the treatment of onchocerciasis and that it appears to be free of major ocular side effects.


Targeting CBX3 with a Dual BET/PLK1 Inhibitor Enhances the Antitumor Efficacy of CDK4/6 Inhibitors in Prostate Cancer.
The development of castration-resistant prostate cancer (CRPC) is a significant factor that reduces life expectancy among patients with prostate cancer. Previously, it is reported that CDK4/6 inhibitors can overcome the resistance of CRPC to BET inhibitors by destabilizing BRD4, suggesting that the combination of CDK4/6 inhibitors and BET inhibitors is a promising approach for treating CRPC. In this study, candidates that affect the combined antitumor effect of CDK4/6 inhibitors and BET inhibitors on CRPC is aimed to examine. The data demonstrates that CBX3 is abnormally upregulated in CDK4/6 inhibitors-resistant cells. CBX3 is almost positively correlated with the cell cycle in multiple malignancies and is downregulated by BET inhibitors. Mechanistically, it is showed that CBX3 is transcriptionally upregulated by BRD4 in CRPC cells. Moreover, it is demonstrated that CBX3 modulated the sensitivity of CRPC to CDK4/6 inhibitors by binding with RB1 to release E2F1. Furthermore, it is revealed that PLK1 phosphorylated CBX3 to enhance the interaction between RB1 and CBX3, and desensitize CRPC cells to CDK4/6 inhibitors. Given that BRD4 regulates CBX3 expression and PLK1 affects the binding between RB1 and CBX3, it is proposed that a dual BRD4/PLK1 inhibitor can increase the sensitivity of CRPC cells to CDK4/6 inhibitors partially through CBX3.


Measurement of vitamin E in serum and plasma by high performance liquid chromatography with electrochemical detection.
A deficiency of vitamin E has been associated with a wide variety of pathological conditions, many of them involving neonates. Previously published methods for the quantitation of vitamin E require large sample volumes as well as extensive extraction and concentration procedures. The present work describes a method for the quantitation of vitamin E in small volumes (less than or equal to 50 microliters) of serum or plasma without the need for extraction or concentration of the sample. With further optimization, even smaller volumes (less than or equal to 10 microliters) can be employed. Sample preparation consisted of precipitation of plasma proteins with absolute ethanol. After centrifugation, a small volume (10 to 50 microliters) of the supernatant was injected directly onto the high performance liquid chromatography (HPLC) column. A dual channel electrochemical detector was used for quantitation of the vitamin E. Pooled serum or plasma was used to determine within-day (8.43 +/- 0.19 mg/L) and between-day (8.49 +/- 0.34 mg/L) precision. Analytical recovery of added vitamin E (4-16 mg/L) averaged 102 +/- 7.0%. No interfering peaks were observed. Linearity was demonstrated over 0.5 to 800 ng. Vitamin E acetate is not detected by this method. HPLC with electrochemical detection is a highly sensitive and specific method for the quantitation of vitamin E in plasma or serum. This procedure is rapid and simple, and can be performed with very small sample volumes.


Adherence to a healthy plant diet may reduce inflammatory factors in obese and overweight women-a cross-sectional study.
Obesity and overweight which are consequence of some interaction factor such as genetics and behavioral habit. Obesity as a metabolic disorder and chronic inflammation is a trigger to countless disease. The main goal of this study is to investigate the interaction of herbal diet on the levels of liver enzymes, inflammatory factors and adipocytes profile. A total of 240 adult women range of 18-48 years were included in the current comparative cross-sectional study. Body composition and dietary intake (using a validated semi-quantitative food frequency questionnaire (FFQ)) were assessed in all participants. In determining a plant based diet index (PDI), vegetarian foods were taken positive score and reverse points for animal foods. For determining a healthful plant based diet index (hPDI), healthy plant foods received positive scores, while less healthy plant foods and animal foods received reverse scores. To create an unhealthful plant-based diet index (uPDI), positive scores were assigned to less healthy plant foods and reverse scores to healthy plant foods and animal foods. For the measurement of serum liver enzymes and inflammatory factors, an enzyme-linked immunosorbent assay (ELISA) method was used. Healthy diet like whole grains, fruits, vegetables, cereals, and beverages such as tea and coffee, based on dietary guidelines, significantly reduced the amount of hs-CRP and TGF-β (P < 0.0001). Higher adhering to hPDI may as a result in higher intake of fiber intake, antioxidants, unsaturated fats, micronutrients, could reduce saturated fats and iron content, and finally weight loss, and reduce inflammation in the body. Base on our finding, in those people who intake higher amounts of healthy plant foods, (vegetable types), instead of unhealthy herbal foods (sweets and desserts), might be useful to reduce inflammation factor like TGF-β and hs-CRP. Women with a higher compliance score in uPDI group (juices, refined grains, starches sweetened with sugar, sweets, and desserts) did not have significantly increase in inflammatory factors levels.


Alpha(2) adrenoceptors regulate proliferation of human intestinal epithelial cells.
Previous studies on rodents have suggested that catecholamines stimulate proliferation of the intestinal epithelium through activation of alpha(2) adrenoceptors located on crypt cells. The occurrence of this effect awaits demonstration in humans and the molecular mechanisms involved have not yet been elucidated. Here, we examined the effect of alpha(2) agonists on a clone of Caco2 cells expressing the human alpha(2A) adrenoceptor. Cells were transfected with a bicistronic plasmid containing the alpha2C10 and neomycin phosphotransferase genes. G418 resistant clones were assayed for receptor expression using radioligand binding. Receptor functionality was assessed by testing its ability to couple Gi proteins and to inhibit cAMP production. Mitogen activated protein kinase (MAPK) phosphorylation was followed by western blot, and cell proliferation was estimated by measuring protein and DNA content. Permanent transfection of Caco2 cells allowed us to obtain a clone (Caco2-3B) expressing alpha(2A) adrenoceptors at a density similar to that found in normal human intestinal epithelium. Caco2-3B retained morphological features and brush border enzyme expression characteristic of enterocytic differentiation. The receptor was coupled to Gi2/Gi3 proteins and its stimulation caused marked diminution of forskolin induced cAMP production. Treatment of Caco2-3B with UK14304 (alpha(2) agonist) induced a rapid increase in the phosphorylation state of MAPK, extracellular regulated protein kinase 1 (Erk1), and 2 (Erk2). This event was totally abolished in pertussis toxin treated cells and in the presence of kinase inhibitors (genistein or PD98059). It was unaffected by protein kinase C downregulation but correlated with a transient increase in Shc tyrosine phosphorylation. Finally, sustained exposure of Caco2-3B to UK14304 resulted in modest but significant acceleration of cell proliferation. None of these effects was observed in the parental cell line Caco2. The results obtained in the present study support a regulatory role for alpha(2) adrenoceptors in intestinal cell proliferation.


Biosynthesis of Rubellins in Ramularia collo-cygni-Genetic Basis and Pathway Proposition.
The important disease Ramularia leaf spot of barley is caused by the fungus Ramularia collo-cygni. The disease causes yield and quality losses as a result of a decrease in photosynthesis efficiency due to the appearance of necrotic spots on the leaf surface. The development of these typical Ramularia leaf spot symptoms is thought to be linked with the release of phytotoxic secondary metabolites called rubellins in the host. However, to date, neither the biosynthetic pathways leading to the production of these metabolites nor their exact role in disease development are known. Using a combined in silico genetic and biochemistry approach, we interrogated the genome of R. collo-cygni to identify a putative rubellin biosynthetic gene cluster. Here we report the identification of a gene cluster containing homologues of genes involved in the biosynthesis of related anthraquinone metabolites in closely related fungi. A putative pathway to rubellin biosynthesis involving the genes located on the candidate cluster is also proposed.


Interaction of corticosterone and gonadal steroids on lipid deposition in the female rat.
The present study was designed to evaluate the interaction of corticosterone (CORT) and female gonadal steroids on energy balance and lipid metabolism. To this end, a 2 x 4 factorial experiment was carried out in which two cohorts of rats differing in their ovary status [OV status: intact (INT) and ovariectomy (OVX)] were each divided into four groups defined by their CORT status [CORT status: nonadrenalectomized (non-ADX), ADX without CORT replacement (placebo subcutaneous pellet), ADX with low-dose CORT replacement, and ADX with high-dose CORT replacement]. After 3 wk of treatment and a 12-h fast, rats were killed and their carcasses analyzed for energy (lipid and protein) content. In addition, indexes of endogenous triglyceride (TRIG) production (liver TRIG content), transport into plasma (triglyceridemia), and incorporation into fat stores [lipoprotein lipase (LPL) activity in adipose tissue (AT)] were assessed. OV and CORT status interacted on body weight gain, total energy, and fat gains. The interactions arose from the fact that the twofold increase in these variables brought on by OVX was abolished by ADX and restored by CORT replacement. Although in ADX groups there was a dose-related restoration of total energy and fat gain by CORT replacement in both INT and OVX cohorts, the impact thereupon of OVX observed in the non-ADX group reappeared only in ADX animals receiving the high dose of CORT. Protein gain was increased by OVX solely in non-ADX rats, whereas the high dose of CORT prevented any net protein gain independently of the OV status. Consistent with treatment effects on total body fat gain, OVX resulted in an increase in liver TRIG content, AT weight, AT LPL activity, and plasma insulin. All these effects of OVX were abolished by ADX and restored by the high dose of CORT. Plasma TRIG were unaffected by OV status but were highly responsive to CORT status. All treatment effects were highly correlated with cumulative food intake. This study shows that the presence of CORT is required for OVX to exert its action on global energy balance and the concomitant, closely integrated adaptations of lipid metabolism.


Beliefs and Emotions Underpin Community Attitudes Towards Voluntary Assisted Dying in Australia.
Assisted dying refers to the steps of the administration of a voluntary assisted dying substance and the administration of the substance. In Australia, assisted dying is now legal in all states. However, there is limited knowledge of what underpins the community's attitudes toward assisted dying. It is important for health professionals to understand what underpins attitudes toward assisted dying when navigating the option with patients and family members. We used a survey based on the Tripartite Model of Attitudes to examine the degree to which beliefs, emotions, and experiences, as well as knowledge, underpinned attitudes towards assisted dying. With a sample of 108, we found emotions and beliefs to significantly explain attitudes towards assisted dying. Knowledge of assisted dying practices was low amongst the sample. Our findings suggest that education of assisted dying is required, and that attitudes towards assisted dying in the Australian community are underpinned by beliefs and emotions.


NTCP model validation method for DAHANCA patient selection of protons versus photons in head and neck cancer radiotherapy.
Introduction: Prediction models using logistic regression may perform poorly in external patient cohorts. However, there is a need to standardize and validate models for clinical use. The purpose of this project was to describe a method for validation of external NTCP models used for patient selection in the randomized trial of protons versus photons in head and neck cancer radiotherapy, DAHANCA 35. Material and methods: Organs at risk of 588 patients treated primarily with IMRT in the randomized controlled DAHANCA19 trial were retrospectively contoured according to recent international recommendations. Dose metrics were extracted using MatLab and all clinical parameters were retrieved from the DAHANCA database. The model proposed by Christianen et al. to predict physician-rated dysphagia was validated through the closed testing, where change of the model intercept, slope and individual beta's were tested for significant prediction improvements. Results: Six months prevalence of dysphagia in the validation cohort was 33%. The closed testing procedure for physician-rated dysphagia showed that the Christianen et al. model needed an intercept refitting for the best match for the Danish patients. The intercept update increased the risk of dysphagia for the validation cohort by 7.9 ± 2.5% point. For the raw model performance, the Brier score (mean squared residual) was 0.467, which improved significantly with a new intercept to 0.415. Conclusions: The previously published Dutch dysphagia model needed an intercept update to match the Danish patient cohort. The implementation of a closed testing procedure on the current validation cohort allows quick and efficient validation of external NTCP models for patient selection in the future.


Soft shoulders ahead: spurious signatures of soft and partial selective sweeps result from linked hard sweeps.
Characterizing the nature of the adaptive process at the genetic level is a central goal for population genetics. In particular, we know little about the sources of adaptive substitution or about the number of adaptive variants currently segregating in nature. Historically, population geneticists have focused attention on the hard-sweep model of adaptation in which a de novo beneficial mutation arises and rapidly fixes in a population. Recently more attention has been given to soft-sweep models, in which alleles that were previously neutral, or nearly so, drift until such a time as the environment shifts and their selection coefficient changes to become beneficial. It remains an active and difficult problem, however, to tease apart the telltale signatures of hard vs. soft sweeps in genomic polymorphism data. Through extensive simulations of hard- and soft-sweep models, here we show that indeed the two might not be separable through the use of simple summary statistics. In particular, it seems that recombination in regions linked to, but distant from, sites of hard sweeps can create patterns of polymorphism that closely mirror what is expected to be found near soft sweeps. We find that a very similar situation arises when using haplotype-based statistics that are aimed at detecting partial or ongoing selective sweeps, such that it is difficult to distinguish the shoulder of a hard sweep from the center of a partial sweep. While knowing the location of the selected site mitigates this problem slightly, we show that stochasticity in signatures of natural selection will frequently cause the signal to reach its zenith far from this site and that this effect is more severe for soft sweeps; thus inferences of the target as well as the mode of positive selection may be inaccurate. In addition, both the time since a sweep ends and biologically realistic levels of allelic gene conversion lead to errors in the classification and identification of selective sweeps. This general problem of "soft shoulders" underscores the difficulty in differentiating soft and partial sweeps from hard-sweep scenarios in molecular population genomics data. The soft-shoulder effect also implies that the more common hard sweeps have been in recent evolutionary history, the more prevalent spurious signatures of soft or partial sweeps may appear in some genome-wide scans.


Spontaneous post-partum cervical carotid artery dissection in a patient with reversible cerebral vasoconstriction syndrome.
Post-partum cervicocephalic artery dissection (pp-CAD) is a rare and poorly understood condition. To our knowledge, only 21 cases have been reported. Reversible cerebral segmental vasoconstriction (RCSV) was first described by Call and Fleming in 1988, and its association with pp-CAD has only been reported in three cases. However, in those cases it is unclear whether the pp-CAD may have been caused by straining during labor and therefore merely coincidental to the intracranial arteriopathy. We describe a 41-year-old right-handed African-American woman who developed the syndrome of pp-CAD (headaches, trace subarachnoid hemorrhage and diffuse cerebral arteriopathy on angiogram) two weeks after delivery. In this unique case, the patient had fortuitously undergone an MR study twice over a four day period which included the carotid bifurcations. During that time the patient was an inpatient, on bed rest and subject to continuous cardiac monitoring. The interval studies documented a true spontaneous right internal carotid artery dissection occurring without obvious cause. The patient had noted moderate right neck pain developing between the two MR studies but experienced no neurological deficits. Subsequent conventional angiography confirmed the presence of postpartum cerebral arteriopathy and the cervical dissection. The patient was managed conservatively with antiplatelet medication and had an otherwise uneventful course. We hypothesize whether transient arterial wall abnormalities, postpartum hormonal changes or subtle connective tissue aberrations play a similar role in the pathogenesis of these two associated conditions.


Three-Dimensional Echocardiographic Assessment of Mitral Annular Physiology in Patients With Degenerative Mitral Valve Regurgitation Undergoing Surgical Repair: Comparison between Early- and Late-Stage Severe Mitral Regurgitation.
Ventricular-annular decoupling is thought to exist in all degenerative myxomatous mitral valve (MV) diseases. However, the annular physiology of degenerative MV disease may differ when severe mitral regurgitation (MR) presents at different stages. The aim of this study was to assess differences in mitral annular physiology and surgical effects between early- and late-stage severe MR. Three-dimensional (3D) transesophageal echocardiography was performed before and after MV surgery in 74 patients with degenerative MV disease, including 57 with early-stage severe MR (without left ventricular remodeling) and 17 with late-stage MR (with left ventricular remodeling). A control group comprised 46 patients without MV disease. Novel 3D MV software was used to evaluate mitral annular dynamics. The degree of annular saddle shape was calculated as the ratio of annular height (AH) to lateromedial diameter (LM). Ventricular-annular decoupling was defined as insufficient systolic AH/LM compared with the control group. Prebypass 3D measurements demonstrated that systolic AH/LM in the early-stage group (0.19 ± 0.04) was similar to that in the control group (0.21 ± 0.05; P = .101), while systolic AH/LM in the late-stage group (0.17 ± 0.04) was lower than that in the control group (P = .011). Postbypass comparison showed saddle shape accentuation in the early-stage group (0.20 ± 0.04), similar to that in the control group (P = .3127); the mitral annulus remained flat in the late-stage group (0.17 ± 0.03; P = .004). Ventricular-annular decoupling, present in the late-stage group, was absent in the early-stage group. MV repair surgery did not disrupt mitral annular saddle shape in the early-stage group; however, it failed to correct annular dysfunction in the late-stage group. Sequential 3D transesophageal echocardiographic analysis provides comprehensive mitral annular evaluation beyond conventional two-dimensional parameters for determining stages of severe MR.


Parallels and differences between innate and adaptive lymphocytes.
Lymphocytes are essential in innate and adaptive immunity. Recent insights suggest that some innate lymphocytes execute functions with adaptive characteristics, while adaptive lymphocytes can operate in ways reminiscent of innate cells. Rather than partitioning lymphocytes according to the type of effector function they execute, we propose that a relevant discrimination relates to the existence of conventional T cells in a naive state. The naive state can be seen as an actively repressed condition that supports T cell diversity and enables the flexible differentiation of effector cells in a manner that best addresses the antigenic challenge. We discuss these considerations in the context of the relative roles of innate lymphoid cells and antigen-experienced T cells in the immune system.


A Comparative Study of Ethanol and Citric Acid Solutions for Extracting Betalains and Total Phenolic Content from Freeze-Dried Beetroot Powder.
This research compares the extraction of betalains (betacyanin and betaxanthin) and total phenolic content using citric acid and aqueous-ethanol solutions. The aim is to find an environmentally sustainable alternative solvent for extracting these compounds from dried beetroot powder. Using citric acid solution as a solvent offers several benefits over ethanol. Citric acid is a weak organic acid found naturally in citrus fruits, making it a safe and environmentally friendly choice for certain extraction processes. Moreover, the use of citric acid as solvent offers biodegradability, non-toxicity, non-flammability, and is cost effective. A full factorial design and response surface methodology (RSM) were employed to assess the effects of extraction parameters (extraction time (5-30 min), extraction temperature (20, 30, 40 °C), pH of citric acid solution (3, 4, 5) and ethanol concentration (10, 20, 30% v/v)). The yield was determined spectrophotometrically and expressed as mg/g of dry powder. The results showed that citric acid solution yielded 85-90% of the ethanolic extract under identical conditions. The maximum yields of betacyanin, betaxanthin, and total phenolic content in citric acid solution were 3.98 ± 0.21 mg/g dry powder, 3.64 ± 0.26 mg/g dry powder, and 8.28 ± 0.34 mg/g dry powder, respectively, while aqueous-ethanol yielded 4.38 ± 0.17 mg/g dry powder, 3.95 ± 0.22 mg/g dry powder, and 8.45 ± 0.45 mg/g dry powder. Optimisation resulted in maximum extraction yields of 90% for betalains and 85% for total phenolic content. The study demonstrates the potential of citric acid as a viable alternative to polar organic solvents for extracting phytochemicals from plant material, providing comparable results to aqueous-ethanol. Artificial Neural Network (ANN) models outperformed RSM in predicting extraction yields. Overall, this research highlights the importance of exploring bio-solvents to enhance the environmental sustainability of phytochemical extraction.


Classification and scoring systems for metastatic spine tumors: a literature review.
Accurate evaluation of metastasis and life prognosis is essential for selecting a suitable therapeutic strategy for metastatic spine tumors owing to limitations in treatment options. For this purpose, various classification, evaluation, and scoring systems have been developed. Classification, evaluation, and scoring systems for metastatic spine tumors reported to date were identified by performing a literature search on PubMed. We reviewed the most cited classifications and scorings before 2009, and all classifications and scorings reported after 2010 from the search results. Six classifications and 23 scorings were reviewed. The classification/evaluation methods are divided into 1) anatomical classification/evaluation methods, 2) evaluation methods for neurological symptoms/instability, and 3) scoring systems for predicting life expectancy. The first 2 were useful for the planning and evaluation of surgical indications. Scoring systems for life prognosis also permitted rough prediction of the outcomes and were useful for the selection of a suitable treatment. However, variation of the patient background, diversity of adopted prognostic factors, and the absence of scoring systems that could predict the outcome with an accuracy of 90% or higher introduced some limitations. The identified classification, evaluation, and scoring systems have been generally useful for treatment strategies. However, we emphasize the necessity of multidisciplinary development and revision of classification and evaluation methods to adapt to the prolongation of survival associated with increased diversity and improvement of treatment options.