Nutrient and Chemical Contaminant Levels in Five Marine Fish Species from Angola-The EAF-Nansen Programme.
Fish is a rich source of several important nutrients and an important part of the otherwise plant-dominated diet present in Angola. However, fish may also be a source of contaminants. The aim of this study was to analyse the nutrient contents and the levels of chemical contaminants, including arsenic, cadmium, mercury, and lead, in five commonly consumed marine fish species sampled during a survey with the research vessel Dr. Fridtjof Nansen in Angola. The species' contribution to recommended nutrient intakes (RNI) for women and children was assessed and compared to that of food products of terrestrial animal origin. All the sampled species are good sources of protein and micronutrients if included in the diet, and inter-species variation is evident. The species were identified to contribute 5-15% of the RNI for calcium, iron, iodine, and zinc and exceeded the contribution to protein and iron intakes of food products of terrestrial animal origin. Furthermore, the potential consumer exposure to chemical contaminants in the species was assessed. None of the species exceeded the maximum levels for cadmium, mercury, and lead, and the potential consumer exposure to cadmium and methylmercury was considered low. The data presented in this study represent an important contribution to African food composition tables.


Adherence to accelerometer protocols among women from economically disadvantaged neighborhoods.
Objective measurement of physical activity with accelerometers is a challenging task in community-based intervention research. Challenges include distribution of and orientation to monitors, nonwear, incorrect placement, and loss of equipment. Data collection among participants from disadvantaged populations may be further hindered by factors such as transportation challenges, competing responsibilities, and cultural considerations. Research staff distributed accelerometers and provided an orientation that was tailored to the population group. General adherence strategies such as follow-up calls, daily diaries, verbal and written instructions, and incentives were accompanied by population-specific strategies such as assisting with transportation, reducing obstacles to wearing the accelerometer, tailoring the message to the participant population, and creating a nonjudgmental environment. Sixty women asked to wear the Actigraph GT1M returned the accelerometer, and 57 of them provided sufficient data for analysis (at least 10 hours a day for a minimum of 4 days) resulting in 95% adherence to the protocol. Participants wore the accelerometers for an average of 5.98 days and 13.15 hours per day. The high accelerometer monitoring adherence among this group of economically disadvantaged women demonstrates that collection of high-quality, objective physical activity data from disadvantaged populations in field-based research is possible.


A comparison of body protein determination in rats by in vivo neutron activation and carcass analysis.
Total body nitrogen (TBN) was determined in 16 rat carcasses ranging in weight from 55 to 550 g, by non-destructive 14 MeV neutron activation analysis (NAA). The rat carcasses were subsequently analysed for TBN by Kjeldahl digestion, for total body water (TBW) by loss of weight after freeze-drying and for body fat by adiabatic bomb calorimetry after subtraction of protein energy. TBN results by the two methods were in good agreement, the precision by NAA (coefficient of variation = 1.5%) being superior to that by chemical analysis (coefficient of variation = 2.8%). Body fat calculated by difference from a combination of measured TBW and NAA data agreed closely with bomb calorimetry measurements. The use of indirect estimates of TBW to determine fat gave poor results. A group of four growing rats was analysed sequentially by NAA four times in 2 weeks. The maximum total radiation dose received by each animal was less than 50 rem (less than 500 mSv) and no significant differences in growth rate were observed compared with non-irradiated control groups. 14 MeV NAA in vivo can be conducted with sufficient precision to measure 0.14 g TBN changes in growing rats at 2 weeks post-weaning.


Non-viral causes of hepatocellular carcinoma.
Hepatocellular carcinoma (HCC) is the most common primary liver malignancy and represents an international public health concern as one of the most deadly cancers worldwide. The main etiology of HCC is chronic infection with hepatitis B and hepatitis C viruses. However, there are other important factors that contribute to the international burden of HCC. Among these are obesity, diabetes, non-alcoholic steatohepatitis and dietary exposures. Emerging evidence suggests that the etiology of many cases of HCC is in fact multifactorial, encompassing infectious etiologies, comorbid conditions and environmental exposures. Clarification of relevant non-viral causes of HCC will aid in preventative efforts to curb the rising incidence of this disease.


Are 'Village Doctors' in Bangladesh a curse or a blessing?
Bangladesh is one of the health workforce crisis countries in the world. In the face of an acute shortage of trained professionals, ensuring healthcare for a population of 150 million remains a major challenge for the nation. To understand the issues related to shortage of health workforce and healthcare provision, this paper investigates the role of various healthcare providers in provision of health services in Chakaria, a remote rural area in Bangladesh. Data were collected through a survey carried out during February 2007 among 1,000 randomly selected households from 8 unions of Chakaria Upazila. Information on health-seeking behaviour was collected from 1 randomly chosen member of a household from those who fell sick during 14 days preceding the survey. Around 44% of the villagers suffered from an illness during 14 days preceding the survey and of them 47% sought treatment for their ailment. 65% patients consulted Village Doctors and for 67% patients Village Doctors were the first line of care. Consultation with MBBS doctors was low at 14%. Given the morbidity level observed during the survey it was calculated that 250 physicians would be needed in Chakaria if the patients were to be attended by a qualified physician. With the current shortage of physicians and level of production in the country it was asserted that it is very unlikely for Bangladesh to have adequate number of physicians in the near future. Thus, making use of existing healthcare providers, such as Village Doctors, could be considered a realistic option in dealing with the prevailing crisis.


Platelet-derived growth factor stabilises vascularisation in collagen-glycosaminoglycan scaffolds in vitro.
Collagen-glycosaminoglycan (CG) scaffolds have been widely developed for a range of regenerative medicine applications. To enhance their efficacy, CG scaffolds have previously been prevascularised in vitro using human umbilical vein endothelial cells and human mesenchymal stromal cells (hMSCs); however, at later timepoints, a regression of vascularisation is observed. This is undesirable for longer preculture periods (e.g., for partial/full organ regeneration) and for in vitro vascularised tissue model systems (e.g., for drug testing/modelling). We hypothesised that delayed platelet-derived growth factor (PDGF)-BB addition could stabilise vessels, preventing their regression. In 2D, we identified 25 ng/ml as a suitable dose that enhanced hMSC metabolic activity and proliferation, without affecting endothelial cells, or migration in either cell type. In our 3D model of CG scaffold vascularisation, early addition of PDGF (Day 3) behaved similarly to no PDGF controls. However, PDGF addition at later timepoints (i.e., Days 4 and 5), with a second addition on Day 10, prevented vascular regression. In quantifying our observations, we identified a need for a tool to measure in vitro vascularisation in porous scaffolds. This was a second key objective of this work. A novel ImageJ macro was developed, which allowed us to analyse vessel-like structures, evaluating their number and morphology, and confirmed our qualitative observations. Finally, upregulation of angiogenic genes (ANG1, KDR, and TEK2) involved in vessel maturation illustrated how PDGF addition contributed to vascular stability. Taken together, the results suggest that addition of PDGF at specific timepoints can be used to stabilise vasculature in CG scaffolds.


The indirect effect of the therapeutic alliance and alcohol abstinence self-efficacy on alcohol use and alcohol-related problems in Project MATCH.
Empirical literature indicates that the therapeutic alliance explains a modest but reliable proportion of variance in predicting alcohol-related outcomes among individuals in treatment for alcohol use disorders (AUDs). Hartzler and colleagues (2011) showed in the COMBINE data set that alcohol abstinence self-efficacy is a potentially important statistical mediator of the relationship between the alliance and client outcomes. The purpose of this study was to replicate this finding in the Project MATCH data set. We used total alliance ratings on the Working Alliance Inventory and tested both client and therapist ratings in mediation analyses. We found that posttreatment self-efficacy accounted for the effect of therapist and client ratings of alliance (measured at session 2) on posttreatment drinking outcomes (drinks per drinking day and alcohol-related problems). In addition, we found a moderation effect of treatment, such that the association between the client's rating of the alliance and self-efficacy changes was positive for individuals in the cognitive behavioral treatment group but negative for those receiving motivation enhancement or Twelve-Step Facilitation. This study reaffirms the importance of the therapeutic alliance and self-efficacy in predicting AUD outcomes. Future research should examine changes in the therapeutic alliance throughout treatment and how these changes are related to self-efficacy and AUD treatment outcomes over time.


Painted black: Iguana melanoderma (Reptilia, Squamata, Iguanidae) a new melanistic endemic species from Saba and Montserrat islands (Lesser Antilles).
The Lesser Antilles, in the Eastern Caribbean, is inhabited by three Iguana species: the Lesser Antillean iguanaIguana delicatissima, which is endemic to the northernmost islands of the Lesser Antilles, the introduced common iguana from South America, Iguana iguana iguana, represented also by the two newly described endemic subspecies Iguana iguana sanctaluciae from Saint Lucia and Iguana iguana insularis from Saint Vincent and the Grenadines, and Grenada, and the introduced Iguana rhinolopha from Central America. Drawing on both morphological and genetic data, this paper describes the Iguana populations from Saba and Montserrat as a new species, Iguana melanoderma. This species is recognized on the basis of the following combination of characteristics: private microsatellite alleles, unique mitochondrial ND4 haplotypes, a distinctive black spot between the eye and tympanum, a dorsal carpet pattern on juveniles and young adults, a darkening of body coloration with aging (except for the anterior part of the snout), a black dewlap, pink on the jowl, the high number of large tubercular nape scales, fewer than ten medium sized-triangular dewlap spikes, high dorsal spikes, and lack of horns on the snout. This new melanistic taxon is threatened by unsustainable harvesting (including for the pet trade) and both competition and hybridization from escaped or released invasive alien iguanas (I. iguana iguana and I. rhinolopha) from South and Central America, respectively. The authors call for action to conserve Iguana melanoderma in Saba and Montserrat and for further research to investigate its relationship to other melanistic iguanas from the Virgin Islands and coastal islands of Venezuela.


A two-dimensional fingerprint nanoprobe based on black phosphorus for bio-SERS analysis and chemo-photothermal therapy.
Flake-shaped nanohybrids based on black phosphorus (BP) have been developed as multifunctional theranostic nanoplatforms for drug delivery, phototherapy and bioimaging. In this work, we report a facile strategy for fabrication of black phosphorus-Au nanoparticle hybrids (BP-AuNPs), which reveal an extraordinary near-infrared (NIR) photothermal transduction efficiency and drug delivery capacity. The applications of the nanocomposites as therapeutic agents for high-performance chemo-photothermal tumor therapy are accomplished in vitro and in vivo. BP-AuNPs also exhibit wonderful surface-enhanced Raman scattering (SERS) activity under NIR laser excitation with a low Raman background, allowing BP-AuNPs to be used as a promising two-dimensional (2D) fingerprint nanoprobe for bio-SERS analysis. The cellular component identification and label-free live-cell bioimaging based on this type of 2D SERS substrate are generally investigated, which open up promising new perspectives in nanomedicine, including diagnosis, imaging and therapy.


Chlorotoxin labeled magnetic nanovectors for targeted gene delivery to glioma.
Glioma accounts for 80% of brain tumors and currently remains one of the most lethal forms of cancers. Gene therapy could potentially improve the dismal prognosis of patients with glioma, but this treatment modality has not yet reached the bedside from the laboratory due to the lack of safe and effective gene delivery vehicles. In this study we investigate targeted gene delivery to C6 glioma cells in a xenograft mouse model using chlorotoxin (CTX) labeled nanoparticles. The developed nanovector consists of an iron oxide nanoparticle core, coated with a copolymer of chitosan, polyethylene glycol (PEG), and polyethylenimine (PEI). Green fluorescent protein (GFP) encoding DNA was bound to these nanoparticles, and CTX was then attached using a short PEG linker. Nanoparticles without CTX were also prepared as a control. Mice bearing C6 xenograft tumors were injected intravenously with the DNA-bound nanoparticles. Nanoparticle accumulation in the tumor site was monitored using magnetic resonance imaging and analyzed by histology, and GFP gene expression was monitored through Xenogen IVIS fluorescence imaging and confocal fluorescence microscopy. Interestingly, the CTX did not affect the accumulation of nanoparticles at the tumor site but specifically enhanced their uptake into cancer cells as evidenced by higher gene expression. These results indicate that this targeted gene delivery system may potentially improve treatment outcome of gene therapy for glioma and other deadly cancers.


Dissociative tendencies and individual differences in high hypnotic suggestibility.
Inconsistencies in the relationship between dissociation and hypnosis may result from heterogeneity among highly suggestible individuals, in particular the existence of distinct highly suggestible subtypes that are of relevance to models of psychopathology and the consequences of trauma. This study contrasted highly suggestible subtypes high or low in dissociation on measures of hypnotic responding, cognitive functioning, and psychopathology. Twenty-one low suggestible (LS), 19 low dissociative highly suggestible (LDHS), and 11 high dissociative highly suggestible (HDHS) participants were administered hypnotic suggestibility scales and completed measures of free recall, working memory capacity, imagery, fantasy-proneness, psychopathology, and exposure to stressful life events. HDHS participants were more responsive to positive and negative hallucination suggestions and experienced greater involuntariness during hypnotic responding. They also exhibited impaired working memory capacity, elevated pathological fantasy and dissociative symptomatology, and a greater incidence of exposure to stressful life events. In contrast, LDHS participants displayed superior object visual imagery. These results provide further evidence for two highly suggestible subtypes: a dissociative subtype characterised by deficits in executive functioning and a predisposition to psychopathology, and a subtype that exhibits superior imagery and no observable deficits in functioning.


Coarse-Grained Simulations Suggest Potential Competing Roles of Phosphoinositides and Amphipathic Helix Structures in Membrane Curvature Sensing of the AP180 N-Terminal Homology Domain.
The generation and sensing of membrane curvature by proteins has become of increasing interest to researchers with multiple mechanisms, from hydrophobic insertion to protein crowding, being identified. However, the role of charged lipids in the membrane curvature-sensing process is still far from understood. Many proteins involved in endocytosis bind phosphatidylinositol 4,5-bisphosphate (PIP2) lipids, allowing these proteins to accumulate at regions of local curvature. Here, using coarse-grained molecular dynamics simulations, we study the curvature-sensing behavior of the ANTH domain, a protein crucial for endocytosis. We selected three ANTH crystal structures containing either an intact, split, or truncated terminal amphipathic helix. On neutral membranes, the ANTH domain has innate curvature-sensing ability. In the presence of PIP2, however, only the domain with an intact helix senses curvature. Our work sheds light on the role of PIP2 and its modulation of membrane curvature sensing by proteins.


Preclinical Pharmacology in the Rhesus Monkey of CW 1759-50, a New Ultra-short Acting Nondepolarizing Neuromuscular Blocking Agent, Degraded and Antagonized by L-Cysteine.
WHAT THIS ARTICLE TELLS US THAT IS NEW: BACKGROUND:: Structure-activity studies were performed to identify a new neuromuscular blocking agent retaining the ultra-short acting characteristics of gantacurium, including degradation and reversal by L-cysteine, but lacking its histaminoid properties in man. CW 1759-50 has emerged from this program. Adduction of CW 1759-50 with L-cysteine was studied by high-performance liquid chromatography and mass spectrometry. Institutional Animal Care and Use Committee-approved comparisons of CW 1759-50 to gantacurium were performed in rhesus monkeys. ED95 for neuromuscular blockade was established. Spontaneous recovery was compared to reversal by L-cysteine in paired studies of boluses or infusions. In addition, changes in mean arterial pressure and heart rate after very large doses of 15 to 60 × ED95 were compared. The half-time of adduction of L-cysteine to CW 1759-50 in vitro was 2.3 min. The ED95 of CW 1759-50 was 0.069 ± 0.02 mg/kg; ED95 of gantacurium was 0.081 ± 0.05 mg/kg (P = 0.006). Duration of action (recovery to 95% twitch height after 98 to 99% blockade) was as follows: CW 1759-50, 8.2 ± 1.5 min; and gantacurium, 7.4 ± 1.9 min; (n = 8 and 9, P = 0.355). Administration of L-cysteine (30 mg/kg) shortened recovery (i.e., induced reversal) from CW 1759-50 after boluses or infusions (P always less than 0.0001). Recovery intervals (5 to 95% twitch) ranged from 6.1 to 6.7 min (and did not differ significantly) after boluses of 0.10 to 0.50 mg/kg, as well as control infusions (P = 0.426 by analysis of variance). Dose ratios comparing changes of 30% in mean arterial pressure or heart rate to ED95 for neuromuscular blockade (ED 30% Δ [mean arterial pressure or heart rate]/ED95) were higher for CW 1759-50 than for gantacurium. CW 1759-50, similar to gantacurium, is an ultra-short acting neuromuscular blocking agent, antagonized by L-cysteine, in the monkey. The circulatory effects, however, are much reduced in comparison with gantacurium, suggesting a trial in humans.


Parallel-coupled dual racetrack silicon micro-resonators for quadrature amplitude modulation.
A parallel-coupled dual racetrack silicon micro-resonator structure is proposed and analyzed for M-ary quadrature amplitude modulation. The over-coupled, critically coupled, and under-coupled scenarios are systematically studied. Simulations indicate that only the over-coupled structures can generate arbitrary M-ary quadrature signals. Analytic study shows that the large dynamic range of amplitude and phase of a modulated over-coupled structure stems from the strong cross-coupling between two resonators, which can be understood through a delicate balance between the direct sum and the "interaction" terms. Potential asymmetries in the coupling constants and quality factors of the resonators are systematically studied. Compensations for these asymmetries by phase adjustment are shown feasible.


Waves and aggregation patterns in myxobacteria.
Under starvation conditions, a population of myxobacteria aggregates to build a fruiting body whose shape is species-specific and within which the cells sporulate. Early in this process, cells often pass through a "ripple phase" characterized by traveling linear, concentric, and spiral waves. These waves are different from the waves observed during slime mold aggregation that depend on diffusible morphogens, because myxobacteria communicate by direct contact. The difference is most dramatic when waves collide: rather than annihilating one another, myxobacterial waves appear to pass through one another unchanged. Under certain conditions, the spacing and location of the nascent fruiting bodies is determined by the wavelength and pattern of the waves. Later in fruiting body development, waves are replaced by streams of cells that circulate around small initial aggregates enlarging and rounding them. Still later, pairs of motile aggregates coalesce to form larger aggregates that develop into fruiting bodies. Here we present a mathematical model that quantitatively explains these wave and aggregation phenomena.


Amiloride reduces portal hypertension in rat liver cirrhosis.
This study aimed to investigate the effect of amiloride on portal hypertension. Amiloride is known to inhibit Na(+)/H(+) exchangers on activated hepatic stellate cells. Liver cirrhosis in rats was induced by bile duct ligation (BDL) or thioacetamide (TAA) administration. The effects of zymosan for Kupffer cell (KC) activation or a thromboxane (TX) analogue (U46619) were tested in isolated perfused livers of cirrhotic rats and in vivo. Downstream mechanisms were investigated using Rho kinase inhibitor (Y-27632) or amiloride. Acute and chronic effects of amiloride and canrenoate on portal pressure were compared in perfused livers and in vivo. TXB(2) efflux was measured by ELISA. The phosphorylation state of moesin (p-moesin) as an indicator of Rho kinase activity and expression of the thromboxane synthase were assessed by western blot analyses. The activity of hepatic stellate cells was analysed by western blot and staining for alpha-smooth muscle actin (alpha-SMA). In BDL rats, KC activation via zymosan increased portal pressure. This was attenuated by the Rho kinase inhibitor Y-27632. Increased thromboxane efflux following zymosan infusion remained unaltered by Y-27632. The infusion of amiloride attenuated zymosan- and U46619-induced increases in portal perfusion pressure. In vivo, direct administration of amiloride, but not of canrenoate, lowered portal pressure. In TAA and BDL rats, treatment with amiloride for 3 days reduced basal portal pressure and KC-induced increases in portal pressure whereas canrenoate had no effect. In livers of amiloride-treated animals, the phosphorylation state of moesin and the number of alpha-SMA positive cells were reduced. Amiloride lowers portal pressure in rat liver cirrhosis by inhibition of intrahepatic vasocontraction. Therefore, patients with cirrhosis and portal hypertension may benefit from amiloride therapy.


Four-dimensional electron microscopy.
The discovery of the electron over a century ago and the realization of its dual character have given birth to one of the two most powerful imaging instruments: the electron microscope. The electron microscope's ability to resolve three-dimensional (3D) structures on the atomic scale is continuing to affect different fields, including materials science and biology. In this Review, we highlight recent developments and inventions made by introducing the fourth dimension of time in electron microscopy. Today, ultrafast electron microscopy (4D UEM) enables a resolution that is 10 orders of magnitude better than that of conventional microscopes, which are limited by the video-camera rate of recording. After presenting the central concept involved, that of single-electron stroboscopic imaging, we discuss prototypical applications, which include the visualization of complex structures when unfolding on different length and time scales. The developed UEM variant techniques are several, and here we illucidate convergent-beam and near-field imaging, as well as tomography and scanning-pulse microscopy. We conclude with current explorations in imaging of nanomaterials and biostructures and an outlook on possible future directions in space-time, 4D electron microscopy.


Fate map of the eye-antennal imaginal disc of the tumorous-head mutant of Drosophila melanogaster.
In specific genetic backgrounds, a mutation in the tuh-3 gene results in the homeotic transformation of head structures to either leg disc derivatives or structures normally found in the extreme posterior end of wild-type animals. The origins of the homeotic structures were mapped to defined positions in the eye-antennal imaginal disc by transplanting abnormal regions of discs isolated from tuh-3 mutants into host mwh;e4 larvae. These metamorphosed implants were removed and differentiated structures were identified. Of 211 successfully recovered implants, 157 gave rise to homeotic tissue: abdominal tergite, male or female external genitalia and/or leg tissue. Transformations to abdominal tergite occurred primarily in cells taken from the eye region of the compound disc. Male and female genitalia arose most often in implants taken from the antennal portion of the disc, although some tissue taken from the lateral region of the eye disc also gave rise to external genitalia. Leg structures came exclusively from implants from the antennal region of the imaginal disc. These results suggest that cells from within specific regions of the eye-antennal compound disc are constrained in their developmental potential. An obvious constraint observed with this mutation is a dorsal/ventral one: Cells from the eye disc, a dorsal structure, primarily gave rise to other dorsal structures, abdominal tergite tissue. Cells from the antennal disc, a ventrally derived structure, primarily gave rise to other ventral structures including genital tissue and distal leg.


Detection and targeting insulin growth factor receptor type 2 (IGF2R) in osteosarcoma PDX in mouse models and in canine osteosarcoma tumors.
Osteosarcoma (OS) represents 3.4% of all childhood cancers with overall survival of 70% not improving in 30 years. The consistent surface overexpression of insulin-like growth factor-2 receptor (IGF2R) has been reported in commercial and patient-derived xenograft (PDX) OS cell lines. We aimed to assess efficacy and safety of treating PDX and commercial OS tumors in mice with radiolabeled antibody to IGF2R and to investigate IGF2R expression on canine OS tumors. IGF2R expression on human commercial lines 143B and SaOS2 and PDX lines OS-17, OS-33 and OS-31 was evaluated by FACS. The biodistribution and microSPECT/CT imaging with 111Indium-2G11 mAb was performed in 143B and OS-17 tumor-bearing SCID mice and followed by radioimmunotherapy (RIT) with 177Lutetium-2G11 and safety evaluation. IGF2R expression in randomly selected canine OS tumors was measured by immunohistochemistry. All OS cell lines expressed IGF2R. Biodistribution and microSPECT/CT revealed selective uptake of 2G11 mAb in 143B and OS-17 xenografts. RIT significantly slowed down the growth of OS-17 and 143B tumors without local and systemic toxicity. Canine OS tumors expressed IGF2R. This study demonstrates the feasibility of targeting IGF2R on OS in PDX and spontaneous canine tumors and sets the stage for further development of RIT of OS using comparative oncology.


Receptive language is associated with visual perception in typically developing children and sensorimotor skills in autism spectrum conditions.
A number of studies have evidenced marked difficulties in language in autism spectrum conditions (ASC). Studies have also shown that language and word knowledge are associated with the same area of brain that is also responsible for visual perception in typically developing (TD) individuals. However, in ASC, research suggests word meaning is mapped differently, on to situational sensorimotor components within the brain. Furthermore, motor coordination is associated with communication skills. The current study explores whether motor coordination and visual perception are impaired in children with ASC, and whether difficulties in coordination and visual perception correlate with receptive language levels. 36 children took part: 18 with ASC and 18 TD children, matched on age and non-verbal reasoning. Both groups completed the Movement ABC, Beery-Buktenica Developmental Test of Visual-Motor Integration, British Picture Vocabulary Scale and Matrices (WASI). Results showed that ASC children scored significantly lower on receptive language, coordination and visual motor integration than the TD group. In the TD group receptive language significantly correlated with visual perception; in the ASC group receptive language significantly correlated with balance. These results imply that sensorimotor skills are associated with the understanding of language in ASC and thus the relationship between sensorimotor experiences and language warrants further investigation.


The role of V-ATPase in neuronal and endocrine systems.
Synaptic vesicles have important roles in the neural transmission at nerve terminals: the storage and the controlled exocytosis of neurotransmitters. At least two different factors are responsible for the concentration process: the vacuolar-type H(+)-ATPase (V-ATPase), establishing an electrochemical gradient of protons, and specific transport systems for transmitters. We will discuss our recent progress on the energy-transducing systems in synaptic vesicles: (1) structural aspects of V-ATPase; (2) energy coupling of transport of transmitters; (3) reconstitution of transporters; (4) effects of neurotoxins and neuron blocking agents; (5) function of synaptic-vesicle-like microvesicles from endocrine tissues.


Thermal hyperbolic metamaterials.
We explore the near-field radiative thermal energy transfer properties of hyperbolic metamaterials. The presence of unique electromagnetic states in a broad bandwidth leads to super-planckian thermal energy transfer between metamaterials separated by a nano-gap. We consider practical phonon-polaritonic metamaterials for thermal engineering in the mid-infrared range and show that the effect exists in spite of the losses, absorption and finite unit cell size. For thermophotovoltaic energy conversion applications requiring energy transfer in the near-infrared range we introduce high temperature hyperbolic metamaterials based on plasmonic materials with a high melting point. Our work paves the way for practical high temperature radiative thermal energy transfer applications of hyperbolic metamaterials.


Mammography: review of the controversy, health disparities, and impact on young african american women.
Ongoing debate about mammography screening for women in their 40s has brought awareness to the opportunities and challenges for achieving optimal breast health in young African American women and in battling health inequities that place them at greater risk for mortality from breast cancer. Despite the screening controversy, a need exists to understand the complex issues related to mammography knowledge, attitudes, and behaviors of young minority women, while empowering them to take an active role in their breast health care. The purpose of this article is to describe the complicated issues related to screening in young African American women within the context of the uncertainty about the evidence surrounding screening practices. Literature was reviewed to garner a comprehensive update of the mammography screening controversy and its impact on mammography practices. Nurses should be aware of the mammography screening controversy and breast cancer risk assessment and how they affect young women's participation in mammography screening. Mammography screening should be a shared decision between the patient and healthcare provider. A better understanding of breast health and its effect on young minority women is needed. Nurses have a prominent role to advocate for, empower, and educate patients as they face the task of deciding whether to begin or continue mammography in their 40s.


Chemoradiation for invasive cervical cancer in elderly patients: outcomes and morbidity.
Age may impact survival and treatment in cervical cancer patients. We sought to determine if treatment and survival were different in elderly patients and whether chemoradiation increased morbidity. We performed a retrospective chart review to identify patients treated with definitive radiation therapy at the University of Iowa Hospitals and Clinics between 1997 and 2001. Three hundred sixty-four patients had a new diagnosis of invasive cervical cancer, of which 150 patients were treated with radiation. We excluded patients treated postoperatively or with palliative intent, leaving 96 patients treated with definitive radiation therapy. Patients were divided into two age categories: elderly (>/=65) and nonelderly (<65). We compared these groups with respect to treatment received, morbidity, and survival. Sixty-nine (72%) women were less than 65 years old, and 27 (28%) women were greater than or equal to 65 years old. Chemoradiation was associated with decreased mortality (P < 0.01). The decrease in mortality did not differ between the two age cohorts (all causes: P = 0.66; cancer specific: P = 0.65), nor was there a difference in the complication rate due to chemoradiation (P = 0.70). Although elderly patients were more likely to be diagnosed with nonsquamous histologies (P < 0.01), their odds of receiving chemoradiation were 0.35 (95% CI: 0.13-0.90) times the odds for nonelderly. Elderly cervical cancer patients more often have nonsquamous histology and are likely to receive only radiation therapy compared to younger patients. Treatment with chemoradiation was associated with similar survival increases in both age cohorts. Complication rates between the two were similar. Chemoradiation should be considered in elderly patients with invasive cervical cancer.


Maintenance clinical trials in bipolar disorder: design implications of the divalproex-lithium-placebo study.
Maintenance studies in bipolar disorder have received increased attention in recent years. The interest is driven by apparent contradictions between results of early placebo-controlled trials of lithium and recent open studies, as well as interest in a new group of drugs with mood-stabilizing properties. The multiple outcome indices that require attention in prophylactic bipolar disorder studies add a dimension not present in acute studies of bipolar disorder. We present the methodology of a recently completed randomized, double-blind, placebo-controlled, parallel-group comparison of divalproex and lithium. We examine the consequences of salient design features, along with their implications for future studies. A fundamental conclusion is that such maintenance studies should be designed and executed to emphasize enrollment of patients with relatively active, severe forms of the illness. This goal is not achieved simply, as inherent features of long-term, placebo-controlled studies drive recruitment and enrollment in the direction of patients with milder forms of bipolar disorder. Attention to the frequency of both manic and depressive episodes and the severity of an index manic episode may aid in the selection of patients most suitable for studies designed to achieve adequate statistical power.


UV photofragmentation dynamics of acetaldehyde cations prepared by single-photon VUV ionization.
Acetaldehyde cations (CH3CHO+) were prepared using single-photon vacuum ultraviolet ionization of CH3CHO in a molecular beam and the fragmentation dynamics explored over the photolysis wavelength range 390-210 nm using velocity-map ion imaging and photofragment yield (PHOFY) spectroscopy. Four fragmentation channels are characterized: CH3CHO+→ C2H3O+ + H (I), CH3CHO+→ HCO+ + CH3 (II), CH3CHO+→ CH3+ + HCO (III), CH3CHO+→ CH4+ + CO (IV). Channels (I), (II), and (IV) are observed across the full photolysis wavelength range while channel (III) is observed only at λ < 317 nm. Maximum fragment ion yields are obtained at ∼250 nm. Ion images were recorded over the range 316-228 nm, which corresponds to initial excitation to the B[combining tilde]2A' and C[combining tilde]2A' states of CH3CHO+. The speed and angular distributions are distinctly different for each detected ion and show evidence of both statistical and dynamical fragmentation pathways. At longer wavelengths, fragmentation via channel (I) leads to modest translational energies (ET), consistent with dissociation over a small barrier and production of highly internally excited CH3CO+. Additional components with EINT greater than the CH3CO+ secondary dissociation threshold appear at shorter wavelengths and are assigned to fragmentation products of vinyl alcohol cation or oxirane cation formed by isomerization of energized CH3CHO+. The ET distribution observed for channel (III) products peaks at zero but is notably colder than that predicted by phase space theory, particularly at longer photolysis wavelengths. The colder-than-statistical ET distributions are attributed to contributions from secondary fragmentation of energized CH3CO+ formed via channel (I), which are attenuated by CH3CHO+ isomerization at shorter wavelengths. Fragmentation via channels (II) and (IV) results in qualitatively similar outcomes, with evidence of isotropic statistical components at low-ET and anisotropic components due to excited state dynamics at higher ET.


Dynamin 2 interacts with α-actinin 4 to drive tumor cell invasion.
The large GTPase Dynamin 2 (Dyn2) is known to increase the invasiveness of pancreatic cancer tumor cells, but the mechanisms by which Dyn2 regulates changes in the actin cytoskeleton to drive cell migration are still unclear. Here we report that a direct interaction between Dyn2 and the actin-bundling protein alpha-actinin (α-actinin) 4 is critical for tumor cell migration and remodeling of the extracellular matrix in pancreatic ductal adenocarcinoma (PDAC) cells. The direct interaction is mediated through the C-terminal tails of both Dyn2 and α-actinin 4, and these proteins interact at invasive structures at the plasma membrane. While Dyn2 binds directly to both α-actinin 1 and α-actinin 4, only the interaction with α-actinin 4 is required to promote tumor cell invasion. Specific disruption of the Dyn2-α-actinin 4 interaction blocks the ability of PDAC cells to migrate in either two dimensions or invade through extracellular matrix as a result of impaired invadopodia stability. Analysis of human PDAC tumor tissue additionally reveals that elevated α-actinin 4 or Dyn2 expression are predictive of poor survival. Overall, these data demonstrate that Dyn2 regulates cytoskeletal dynamics, in part, by interacting with the actin-binding protein α-actinin 4 during tumor cell invasion.


Importance of mind-muscle connection during progressive resistance training.
This study evaluates whether focusing on using specific muscles during bench press can selectively activate these muscles. Altogether 18 resistance-trained men participated. Subjects were familiarized with the procedure and performed one-maximum repetition (1RM) test during the first session. In the second session, 3 different bench press conditions were performed with intensities of 20, 40, 50, 60 and 80 % of the pre-determined 1RM: regular bench press, and bench press focusing on selectively using the pectoralis major and triceps brachii, respectively. Surface electromyography (EMG) signals were recorded for the triceps brachii and pectoralis major muscles. Subsequently, peak EMG of the filtered signals were normalized to maximum maximorum EMG of each muscle. In both muscles, focusing on using the respective muscles increased muscle activity at relative loads between 20 and 60 %, but not at 80 % of 1RM. Overall, a threshold between 60 and 80 % rather than a linear decrease in selective activation with increasing intensity appeared to exist. The increased activity did not occur at the expense of decreased activity of the other muscle, e.g. when focusing on activating the triceps muscle the activity of the pectoralis muscle did not decrease. On the contrary, focusing on using the triceps muscle also increased pectoralis EMG at 50 and 60 % of 1RM. Resistance-trained individuals can increase triceps brachii or pectarilis major muscle activity during the bench press when focusing on using the specific muscle at intensities up to 60 % of 1RM. A threshold between 60 and 80 % appeared to exist.


Trench Inserts as Long-term Barriers to Root Transmission for Control of Oak Wilt.
Physical and chemical barriers to root penetration and root grafting across trenches were evaluated for their effectiveness in improving trenches as barriers to root transmission of the oak wilt fungus in live oaks. Four trench insert materials were tested, including water-permeable Typar and Biobarrier, and water-impermeable Geomembrane of two thicknesses. Systemic fungicide treatments of trees immediately outside of trenches also were tested. In the first several years following trench installation, an abundance of small adventitious roots commonly formed from roots severed by trenching. These roots provided opportunities for initiation of root grafts across trenches in subsequent years. Although trench inserts did not significantly improve trenches during the first 3 years following trench installation, water-permeable inserts did effectively improve the performance of trenches beyond the third posttrenching year, when trenches are normally effective, and extended trench longevity indefinitely. The water-permeable inserts were more effective root barriers because they did not direct root growth from the point of root contact. The water-impermeable materials, however, did tend to direct root growth around these barriers, leading to the development of new root graft connections and associated oak wilt root transmission across the trench. The additional cost of trench inserts above trenching costs was justified in urban and rural homestead sites, where high-value landscape trees required more protection and additional retrenching costs were avoided.


Scanning protocols dedicated to smart velocity ranging in spectral OCT.
We introduce a new type of scanning protocols, called segmented protocols, which enable extracting multi-range flow velocity information from a single Spectral OCT data set. The protocols are evaluated using a well defined flow in a glass capillary. As an example of in vivo studies, we demonstrate two- and three-dimensional imaging of the retinal vascular system in the eyes of healthy volunteers. The flow velocity detection is performed using a method of Joint Spectral and Time domain OCT. Velocity ranging is demonstrated in imaging of retinal vasculature in the macular region and in the optic disk area characterized by different flow velocity values. Additionally, an enhanced visualization of retinal capillary network is presented in the close proximity to macula.


Long-read sequencing uncovers the adaptive topography of a carnivorous plant genome.
Utricularia gibba, the humped bladderwort, is a carnivorous plant that retains a tiny nuclear genome despite at least two rounds of whole genome duplication (WGD) since common ancestry with grapevine and other species. We used a third-generation genome assembly with several complete chromosomes to reconstruct the two most recent lineage-specific ancestral genomes that led to the modern U. gibba genome structure. Patterns of subgenome dominance in the most recent WGD, both architectural and transcriptional, are suggestive of allopolyploidization, which may have generated genomic novelty and led to instantaneous speciation. Syntenic duplicates retained in polyploid blocks are enriched for transcription factor functions, whereas gene copies derived from ongoing tandem duplication events are enriched in metabolic functions potentially important for a carnivorous plant. Among these are tandem arrays of cysteine protease genes with trap-specific expression that evolved within a protein family known to be useful in the digestion of animal prey. Further enriched functions among tandem duplicates (also with trap-enhanced expression) include peptide transport (intercellular movement of broken-down prey proteins), ATPase activities (bladder-trap acidification and transmembrane nutrient transport), hydrolase and chitinase activities (breakdown of prey polysaccharides), and cell-wall dynamic components possibly associated with active bladder movements. Whereas independently polyploid Arabidopsis syntenic gene duplicates are similarly enriched for transcriptional regulatory activities, Arabidopsis tandems are distinct from those of U. gibba, while still metabolic and likely reflecting unique adaptations of that species. Taken together, these findings highlight the special importance of tandem duplications in the adaptive landscapes of a carnivorous plant genome.


Soil nitrous oxide flux following land-use reversion from Miscanthus and SRC willow to perennial ryegrass.
Decarbonization of the world's energy supply is essential to meet the targets of the 2016 Paris climate change agreement. One promising opportunity is the utilization of second generation, low input bioenergy crops such as Miscanthus and Short Rotation Coppice (SRC) willow. Research has previously been carried out on the greenhouse gas (GHG) balance of growing these feedstocks and land-use changes involved in converting conventional cropland to their production; however, there is almost no body of work understanding the costs associated with their end of life transitions back to conventional crops. It is likely that it is during crop interventions and land-use transitions that significant GHG fluxes might occur. Therefore, in this study, we investigated soil GHG fluxes over 82 weeks during transition from Miscanthus and SRC willow into perennial ryegrass in west Wales, UK. This study captured soil GHG fluxes at a weekly time step, alongside monthly changes in soil nitrogen and labile carbon and reports the results of regression modelling of suspected drivers. Methane fluxes were typically trivial; however, nitrous oxide (N2O) fluxes were notably affected, reverted plots produced significantly more N2O than retained controls and Miscanthus produced significantly higher fluxes overall than willow plots. N2O costs of reversion appeared to be contained within the first year of reversion when the Miscanthus plots produced an average pregrass flux of 0.13 mg N2O m-2 hr-1 while for willow, this was 0.03 mg N2O m-2 hr-1. Total N2O emission from reversion increased the carbon cost over the lifetime of the Miscanthus from 6.50 to 9.91 Mg CO2 eq. ha-1 while for the willow, this increase was from 9.61 to 10.42 Mg CO2 eq. ha-1. Despite these significant increases, the carbon cost of energy contained in these perennial crops remained far lower than the equivalent carbon cost of energy in coal.


Deaf children with autism spectrum disorders.
Epidemiological studies investigating the prevalence of autism have increased in recent years, within the United States and abroad. However, statistics as to how many of those children may also have a comorbid hearing loss is lacking. The prevalence of school-administrator reported diagnosis of autism spectrum disorders (clinical diagnosis [DSM-IV] and/or IDEA classification) among children with hearing loss in the US was estimated from the 2009–2010 Annual Survey of Deaf and Hard of Hearing Children and Youth conducted by the Gallaudet Research Institute. Results indicate that during the 2009–2010 school year 1 in 59 children (specifically 8-year olds) with hearing loss were also receiving services for autism; considerably higher, than reported national estimates of 1 in 91 (Koganet al. in Pediatrics 124(4):1–8, 2009) and 1 in 110 (CDC 2007) for hearing children. Significantly more children with profound hearing loss had a comorbid diagnosis of autism than those with milder forms of hearing loss. These results are discussed, while highlighting the need for increased awareness and research in a population that has thus far received little services or attention.


Cell surface trafficking of the antidepressant-sensitive norepinephrine transporter revealed with an ectodomain antibody.
The antidepressant-sensitive L-norepinephrine (NE) transporter (NET;SLC6A2) is a critical determinant of neurotransmitter inactivation following NE release at synapses. Although regulated trafficking of NET has been documented in transfected cells, a lack of reagents suitable for reporting native NET surface exposition has limited validation of this concept in neurons. In the current report, we document the utility of a novel antibody (43408) directed at conserved sequences in the NET second extracellular loop. Using human NET (hNET) stably transfected cells, we document loss of NET surface expression following acute (30 min) phorbol ester treatments. In superior cervical ganglion (SCG) cultures, NET surface expression is prominent on varicosities defined by FM1-43 labeling of living neurons or synaptophysin labeling of fixed preparations. Moreover, NET surface density can be rapidly augmented by brief depolarization (5 min, 40 mM K(+)). Similarly, in brainstem cultures, we demonstrate an increase in NET surface labeling following either depolarization or angiotensin II stimulation. These findings provide the first evidence for regulated trafficking of NET in neurons and support the suggestion that activity-dependent NET trafficking may provide additional modulatory capacity for noradrenergic signaling.


Prediction of nonSVR to therapy with pegylated interferon-alpha2a and ribavirin in chronic hepatitis C genotype 1 patients after 4, 8 and 12 weeks of treatment.
In patients with chronic hepatitis C genotype 1, the current algorithm for treatment discontinuation is based on no early virological response (<2 log decline in hepatitis C virus (HCV)-RNA) at 12 weeks. It is important to determine whether prediction of nonsustained virological response (NR) before 12 weeks can be robustly obtained by statistical methods. We used longitudinal discriminant analysis (LDA) to build and cross-validate models including baseline patient characteristics and measurements of serum HCV-RNA in the first 4, 8 or 12 weeks of treatment. The performance of each model was evaluated by the partial AUC (PA) index, exploring the accuracy of prediction in the range of high negative predictive values. Models were compared by computing 95% confidence intervals for the difference in PA indices. NR was best predicted before week 12 by a single HCV-RNA measurement at week 8 taken together with gender, BMI and age (W8 model, PA index = 0.857). This model was not inferior to models that included a measurement at week 12 (PA index = 0.831). The best model obtained with LDA within the first 4 weeks, which included measurements at days 4, 8 and at week 4, was found to be inferior to the week 8 model (PA index = 0.796). These results indicate that lack of sustained viral response is best predicted after 8 weeks of treatment and that waiting until 12 weeks does not improve the prediction.


Purification and characterization of the [NiFe]-hydrogenase of Shewanella oneidensis MR-1.
Shewanella oneidensis MR-1 possesses a periplasmic [NiFe]-hydrogenase (MR-1 [NiFe]-H(2)ase) that has been implicated in H(2) production and oxidation as well as technetium [Tc(VII)] reduction. To characterize the roles of MR-1 [NiFe]-H(2)ase in these proposed reactions, the genes encoding both subunits of MR-1 [NiFe]-H(2)ase were cloned and then expressed in an MR-1 mutant without hyaB and hydA genes. Expression of recombinant MR-1 [NiFe]-H(2)ase in trans restored the mutant's ability to produce H(2) at 37% of that for the wild type. Following purification, MR-1 [NiFe]-H(2)ase coupled H(2) oxidation to reduction of Tc(VII)O(4)(-) and methyl viologen. Change of the buffers used affected MR-1 [NiFe]-H(2)ase-mediated reduction of Tc(VII)O(4)(-) but not methyl viologen. Under the conditions tested, all Tc(VII)O(4)(-) used was reduced in Tris buffer, while in HEPES buffer, only 20% of Tc(VII)O(4)(-) was reduced. The reduced products were soluble in Tris buffer but insoluble in HEPES buffer. Transmission electron microscopy analysis revealed that Tc precipitates reduced in HEPES buffer were aggregates of crystallites with diameters of ∼5 nm. Measurements with X-ray absorption near-edge spectroscopy revealed that the reduction products were a mixture of Tc(IV) and Tc(V) in Tris buffer but only Tc(IV) in HEPES buffer. Measurements with extended X-ray adsorption fine structure showed that while the Tc bonding environment in Tris buffer could not be determined, the Tc(IV) product in HEPES buffer was very similar to Tc(IV)O(2)·nH(2)O, which was also the product of Tc(VII)O(4)(-) reduction by MR-1 cells. These results shows for the first time that MR-1 [NiFe]-H(2)ase catalyzes Tc(VII)O(4)(-) reduction directly by coupling to H(2) oxidation.


Lineage relationship analysis of RORgammat+ innate lymphoid cells.
Lymphoid tissue-inducer (LTi) cells initiate the development of lymphoid tissues through the activation of local stromal cells in a process similar to inflammation. LTi cells express the nuclear hormone receptor RORγt, which also directs the expression of the proinflammatory cytokine interleukin-17 in T cells. We show here that LTi cells are part of a larger family of proinflammatory RORγt(+) innate lymphoid cells (ILCs) that differentiate from distinct fetal liver RORγt(+) precursors. The fate of RORγt(+) ILCs is determined by mouse age, and after birth, favors the generation of cells involved in intestinal homeostasis and defense. Contrary to RORγt(+) T cells, however, RORγt(+) ILCs develop in the absence of microbiota. Our study indicates that RORγt(+) ILCs evolve to preempt intestinal colonization by microbial symbionts.


Mycophenolate mofetil for induction and maintenance of remission in microscopic polyangiitis with mild to moderate renal involvement--a prospective, open-label pilot trial.
Microscopic polyangiitis (MPA) is a systemic small-vessel vasculitis associated with anti-neutrophil cytoplasmic antibodies (ANCA), often targeting myeloperoxidase (MPO). Cyclophosphamide (CYC) plus corticosteroids (CS) is considered standard therapy for patients with renal involvement, but treatment response is not satisfactory in all patients and CYC has well recognized toxicity. This prospective pilot trial explored whether mycophenolate mofetil (MMF) represents an effective alternative to CYC for induction and maintenance of remission in MPA with mild to moderate renal involvement. Seventeen P-ANCA/MPO-ANCA-positive patients with MPA with mild to moderate renal involvement received MMF (1000 mg orally, twice daily) and CS (intravenous methylprednisolone, 1 to 3 g, followed by oral prednisone at 1 mg/kg per day). Oral CS were discontinued by month 6; MMF was continued through month 18. The primary outcome measure was remission by month 6 and stable renal function. Secondary endpoints included major relapses necessitating a switch to CYC plus CS, minor relapses requiring an increase in CS dosage, and adverse events. Thirteen of 17 patients enrolled achieved the primary outcome, and 4 failed because of insufficient response, relapse, or MMF intolerance. Twelve patients remained in remission through month 18, renal function remained stable, and proteinuria improved. Side effects of MMF were mild, transient, and responsive to dose adjustments in all patients except one. MMF represents an alternative to CYC for induction and maintenance of remission in patients with MPO-ANCA-associated MPA with mild to moderate renal disease.


A phase 3 trial of RTS,S/AS01 malaria vaccine in African infants.
The candidate malaria vaccine RTS,S/AS01 reduced episodes of both clinical and severe malaria in children 5 to 17 months of age by approximately 50% in an ongoing phase 3 trial. We studied infants 6 to 12 weeks of age recruited for the same trial. We administered RTS,S/AS01 or a comparator vaccine to 6537 infants who were 6 to 12 weeks of age at the time of the first vaccination in conjunction with Expanded Program on Immunization (EPI) vaccines in a three-dose monthly schedule. Vaccine efficacy against the first or only episode of clinical malaria during the 12 months after vaccination, a coprimary end point, was analyzed with the use of Cox regression. Vaccine efficacy against all malaria episodes, vaccine efficacy against severe malaria, safety, and immunogenicity were also assessed. The incidence of the first or only episode of clinical malaria in the intention-to-treat population during the 14 months after the first dose of vaccine was 0.31 per person-year in the RTS,S/AS01 group and 0.40 per person-year in the control group, for a vaccine efficacy of 30.1% (95% confidence interval [CI], 23.6 to 36.1). Vaccine efficacy in the per-protocol population was 31.3% (97.5% CI, 23.6 to 38.3). Vaccine efficacy against severe malaria was 26.0% (95% CI, -7.4 to 48.6) in the intention-to-treat population and 36.6% (95% CI, 4.6 to 57.7) in the per-protocol population. Serious adverse events occurred with a similar frequency in the two study groups. One month after administration of the third dose of RTS,S/AS01, 99.7% of children were positive for anti-circumsporozoite antibodies, with a geometric mean titer of 209 EU per milliliter (95% CI, 197 to 222). The RTS,S/AS01 vaccine coadministered with EPI vaccines provided modest protection against both clinical and severe malaria in young infants. (Funded by GlaxoSmithKline Biologicals and the PATH Malaria Vaccine Initiative; RTS,S ClinicalTrials.gov number, NCT00866619.).


PRIASE 2021 guidelines for reporting animal studies in Endodontology: a consensus-based development.
Animal testing is crucial in situations when research on humans is not allowed because of unknown health risks and ethical concerns. The current project aims to develop reporting guidelines exclusively for animal studies in Endodontology, using an established consensus-based methodology. The guidelines have been named: Preferred Reporting Items for Animal Studies in Endodontology (PRIASE) 2021. Nine individuals (PD, VN, AK, PM, MN, JF, EP, JJ and SJ), including the project leaders (PD, VN) formed a steering committee. The steering committee developed a novel checklist by adapting and integrating their animal testing and peer review experience with the Animals in Research: Reporting In Vivo Experiments (ARRIVE) guidelines and also the Clinical and Laboratory Images in Publications (CLIP) principles. A PRIASE Delphi Group (PDG) and PRIASE Online Meeting Group (POMG) were also formed. Thirty-one PDG members participated in the online Delphi process and achieved consensus on the checklist items and flowchart that were used to formulate the PRIASE guidelines. The novel PRIASE 2021 guidelines were discussed with the POMG on 9 September 2020 via a Zoom online video call attended by 21 individuals from across the globe and seven steering committee members. Following the discussions, the guidelines were modified and then piloted by several authors whilst writing a manuscript involving research on animals. The PRIASE 2021 guidelines are a checklist consisting of 11 domains and 43 individual items together with a flowchart. The PRIASE 2021 guidelines are focused on improving the methodological principles, reproducibility and quality of animal studies in order to enhance their reliability as well as repeatability to estimate the effects of endodontic treatments and usefulness for guiding future clinical studies on humans.


Deaths of two hospital inpatients poisoned by pilocarpine.
Two inpatients of one hospital ward died. Pilocarpine poisoning was suspected and subsequently confirmed by analysis of urine. The circumstantial evidence strongly suggested that the food given to the patients in the ward had been adulterated. Police inquiries failed to elicit any further information, and open verdicts were returned at the inquest. Precautions taken subsequently to prevent a similar event--sealing food containers and trolleys--entailed a capital cost of 43,000 pounds. In addition, food stores were kept locked and tighter control kept on drugs stored in ward pharmacies.


LSD-induced entropic brain activity predicts subsequent personality change.
Personality is known to be relatively stable throughout adulthood. Nevertheless, it has been shown that major life events with high personal significance, including experiences engendered by psychedelic drugs, can have an enduring impact on some core facets of personality. In the present, balanced-order, placebo-controlled study, we investigated biological predictors of post-lysergic acid diethylamide (LSD) changes in personality. Nineteen healthy adults underwent resting state functional MRI scans under LSD (75µg, I.V.) and placebo (saline I.V.). The Revised NEO Personality Inventory (NEO-PI-R) was completed at screening and 2 weeks after LSD/placebo. Scanning sessions consisted of three 7.5-min eyes-closed resting-state scans, one of which involved music listening. A standardized preprocessing pipeline was used to extract measures of sample entropy, which characterizes the predictability of an fMRI time-series. Mixed-effects models were used to evaluate drug-induced shifts in brain entropy and their relationship with the observed increases in the personality trait openness at the 2-week follow-up. Overall, LSD had a pronounced global effect on brain entropy, increasing it in both sensory and hierarchically higher networks across multiple time scales. These shifts predicted enduring increases in trait openness. Moreover, the predictive power of the entropy increases was greatest for the music-listening scans and when "ego-dissolution" was reported during the acute experience. These results shed new light on how LSD-induced shifts in brain dynamics and concomitant subjective experience can be predictive of lasting changes in personality. Hum Brain Mapp 37:3203-3213, 2016. © 2016 Wiley Periodicals, Inc.


Serum potassium concentration as a risk factor of ventricular arrhythmias early in acute myocardial infarction.
Sixty patients with a first acute myocardial infarction and no current treatment with cardioactive drugs were included in a prospective study of the relationship between serum potassium concentration and the early occurrence of ventricular tachycardia and premature ventricular contractions (PVCs). Serum potassium level (range 2.5 to 5 mmol/liter) was estimated 3.8 +/- 2.5 hr (mean +/- SD) after the onset of the infarction, and Holter monitoring was performed during the subsequent 12 hr. In multivariate analysis, serum potassium level was negatively and age positively related to ventricular tachycardia. Among the subclasses of PVCs (frequent unifocal, multifocal, couplets, bigeminy), serum potassium concentration was negatively related to the frequent unifocal subclass; hypertension was related to couplets and to the presence of any of the subclasses, and serum aspartate aminotransferase concentration was related to multifocal PVCs. Heart failure leading to death was related to all subclasses of PVC. Serum potassium concentration is an independent inverse predictor of the occurrence of ventricular tachycardia and frequent unifocal PVCs early in acute myocardial infarction.


Extreme CD8 T cell requirements for anti-malarial liver-stage immunity following immunization with radiation attenuated sporozoites.
Radiation-attenuated Plasmodium sporozoites (RAS) are the only vaccine shown to induce sterilizing protection against malaria in both humans and rodents. Importantly, these "whole-parasite" vaccines are currently under evaluation in human clinical trials. Studies with inbred mice reveal that RAS-induced CD8 T cells targeting liver-stage parasites are critical for protection. However, the paucity of defined T cell epitopes for these parasites has precluded precise understanding of the specific characteristics of RAS-induced protective CD8 T cell responses. Thus, it is not known whether quantitative or qualitative differences in RAS-induced CD8 T cell responses underlie the relative resistance or susceptibility of immune inbred mice to sporozoite challenge. Moreover, whether extraordinarily large CD8 T cell responses are generated and required for protection following RAS immunization, as has been described for CD8 T cell responses following single-antigen subunit vaccination, remains unknown. Here, we used surrogate T cell activation markers to identify and track whole-parasite, RAS-vaccine-induced effector and memory CD8 T cell responses. Our data show that the differential susceptibility of RAS-immune inbred mouse strains to Plasmodium berghei or P. yoelii sporozoite challenge does not result from host- or parasite-specific decreases in the CD8 T cell response. Moreover, the surrogate activation marker approach allowed us for the first time to evaluate CD8 T cell responses and protective immunity following RAS-immunization in outbred hosts. Importantly, we show that compared to a protective subunit vaccine that elicits a CD8 T cell response to a single epitope, diversifying the targeted antigens through whole-parasite RAS immunization only minimally, if at all, reduced the numerical requirements for memory CD8 T cell-mediated protection. Thus, our studies reveal that extremely high frequencies of RAS-induced memory CD8 T cells are required, but may not suffice, for sterilizing anti-Plasmodial immunity. These data provide new insights into protective CD8 T cell responses elicited by RAS-immunization in genetically diverse hosts, information with relevance to developing attenuated whole-parasite vaccines.


Approximate multiple protein structure alignment using the sum-of-pairs distance.
An algorithm is presented to compute a multiple structure alignment for a set of proteins and to generate a consensus (pseudo) protein for the set. The algorithm is a heuristic in that it computes an approximation to the optimal multiple structure alignment that minimizes the sum of the pairwise distances between the protein structures. The algorithm chooses an input protein as the initial consensus and computes a correspondence between the protein structures (which are represented as sets of unit vectors) using an approach analogous to the center-star method for multiple sequence alignment. From this correspondence, a set of rotation matrices (optimal for the given correspondence) is derived to align the structures and derive the new consensus. The process is iterated until the sum of pairwise distances converges. The computation of the optimal rotations is itself an iterative process that both makes use of the current consensus and generates simultaneously a new one. This approach is based on an interesting result that allows the sum of all pairwise distances to be represented compactly as distances to the consensus. Experimental results on several protein families are presented, showing that the algorithm converges quite rapidly.


Prioritization in visual search: visual marking is not dependent on a mnemonic search.
Visual marking (VM) refers to our ability to completely exclude old items from search when new stimuli are presented in our visual field. We examined whether this ability reflects an attentional scan of the old items, possibly allowing observers to apply inhibition of return or maintain a memory representation of already seen locations. In four experiments, we compared performance in two search conditions. In the double-search (DS) condition, we required participants to pay attention to a first set of items by having them search for a target within the set. Subsequently, they had to search a second set while the old items remained in the field. In the VM condition, the participants expected the target only to be in the second (new) set. Selection of new items in the DS condition was relatively poor and was always worse than would be expected if only the new stimuli had been searched. In contrast, selection of the new items in the VM condition was good and was equal to what would be expected if there had been an exclusive search of the new stimuli. These results were not altered when differences in Set 1 difficulty, task switching, and response generation were controlled for. We conclude that the mechanism of VM is distinct from mnemonic and/or serial inhibition-of-return processes as involved in search, although we also discuss possible links to more global and flexible inhibition-of-return processes not necessarily related to search.


Aggregating public preferences for healthcare: putting theory into practice.
The aim of this study was to investigate alternative methods for aggregating public preferences that may then be used to guide priority-setting decisions in healthcare. This involved asking members of the Australian public to assume the role of a healthcare planner and to allocate an additional (but fixed) amount of healthcare resources across three sets of options: medical/surgical procedures, public health programmes and across population groups exhibiting different characteristics. Two methods of aggregation were used: rank ordering and what the author describes as the 'pie method'. In contrast to conventional social welfare functions (that make use of ordinal, non-comparable utility information of individual consumers), the methods used in this study allowed for additional information on the intensity and the comparability of the preferences. The impact of these different methods on aggregation results is investigated. It is argued that those decision makers charged with the day-to-day responsibility of engaging the general public in resource allocation decision making need to be aware that different methods of aggregation can result in different healthcare priorities. Implications for researchers, including the impact of different forms of information on the formation of social preferences, are also discussed.


The implementation of omics technologies in cancer microbiome research.
Whilst the interplay between host genetics and the environment plays a pivotal role in the aetiopathogenesis of cancer, there are other key contributors of importance as well. One such factor of central and growing interest is the contribution of the microbiota to cancer. Even though the field is only a few years old, investigation of the 'cancer microbiome' has already led to major advances in knowledge of the basic biology of cancer risk and progression, opened novel avenues for biomarkers and diagnostics, and given a better understanding of mechanisms underlying response to therapy. Recent developments in microbial DNA sequencing techniques (and the bioinformatics required for analysis of these datasets) have allowed much more in-depth profiling of the structure of microbial communities than was previously possible. However, for more complete assessment of the functional implications of microbial changes, there is a growing recognition of the importance of the integration of microbial profiling with other omics modalities, with metabonomics (metabolite profiling) and proteomics (protein profiling) both gaining particular recent attention. In this review, we give an overview of some of the key scientific techniques being used to unravel the role of the cancer microbiome. We have aimed to highlight practical aspects related to sample collection and preparation, choice of the modality of analysis, and examples of where different omics technologies have been complementary to each other to highlight the significance of the cancer microbiome.


Mechanical properties and in vitro biological response to porous titanium alloys prepared for use in intervertebral implants.
The generation of titanium foams is a promising strategy for modifying the mechanical properties of intervertebral reinforcements. Thus, the aim of this study was to compare the in vitro biological response of Ti6Al4V alloys with different pore sizes for use in intervertebral implants in terms of the adhesion, proliferation, and differentiation of pre-osteoblastic cells. We studied the production of Ti6Al4V foams by powder metallurgy and the biological responses to Ti6Al4V foams were assessed in terms of different pore interconnectivities and elastic moduli. The Ti6Al4V foams obtained had similar porosities of approximately 34%, but different pore sizes (66 µm for fine Ti6Al4V and 147 µm for coarse Ti6Al4V) due to the sizes of the microsphere used. The Ti6Al4V foams had a slightly higher Young׳s modulus compared with cancellous bone. The dynamic mechanical properties of the Ti6Al4V foams were slightly low, but these materials can satisfy the requirements for intervertebral prosthesis applications. The cultured cells colonized both sizes of microspheres near the pore spaces, where they occupied almost the entire area of the microspheres when the final cell culture time was reached. No statistical differences in cell proliferation were observed; however, the cells filled the pores on fine Ti6Al4V foams but they only colonized the superficial microspheres, whereas the cells did not fill the pores on coarse Ti6Al4V foams but they were distributed throughout most of the material. In addition, the microspheres with wide pores (coarse Ti6Al4V) stimulated higher osteoblast differentiation, as demonstrated by the Alcaline Phosphatase (ALP) activity. Our in vitro results suggest that foams with wide pore facilitate internal cell colonization and stimulate osteoblast differentiation.


The Patient-Provider Relationship Is Associated with Hepatitis C Treatment Eligibility: A Prospective Mixed-Methods Cohort Study.
Hepatitis C virus (HCV) treatment has the potential to cure the leading cause of cirrhosis and hepatocellular carcinoma. However, only those deemed eligible for treatment have the possibility of this cure. Therefore, understanding the determinants of HCV treatment eligibility is critical. Given that effective communication with and trust in healthcare providers significantly influences treatment eligibility decisions in other diseases, we aimed to understand patient-provider interactions in the HCV treatment eligibility process. This prospective cohort study was conducted in the VA Pittsburgh Healthcare System. Patients were recruited after referral for gastroenterology consultation for HCV treatment with interferon and ribavirin. Consented patients completed semi-structured interviews and validated measures of depression, substance and alcohol use, and HCV knowledge. Two coders analyzed the semi-structured interviews. Factors associated with patient eligibility for interferon-based therapy were assessed using multivariate logistic regression. Of 339 subjects included in this analysis, only 56 (16.5%) were deemed eligible for HCV therapy by gastroenterology (GI) providers. In the multivariate logistic regression, patients who were older (OR = 0.96, 95%CI = 0.92-0.99, p = .049), reported concerns about the GI provider (OR = 0.40, 95%CI = 0.10-0.87, p = 0.02) and had depression symptoms (OR = 0.32, 95%CI = 0.17-0.63, p = 0.001) were less likely to be eligible. Patients described barriers that included feeling stigmatized and poor provider interpersonal or communication skills. In conclusion, we found that patients' perceptions of the relationship with their GI providers were associated with treatment eligibility. Establishing trust and effective communication channels between patients and providers may lower barriers to potential HCV cure.


Social deprivation and mortality in adults with diabetes mellitus.
To investigate the relationship between measures of social deprivation and mortality in adults with diabetes, data from 2104 randomly selected adults (> 16 years of age) with Type 1 and Type 2 diabetes mellitus from 8 hospital out-patient departments were analysed. A total of 38% of subjects had Type 1 (diagnosed before the age of 36 years and treated with insulin), 55% were male and 85% Caucasian. During a follow-up period (mean (SD) of 8.4 (0.9) years), 293 (14%) of the subjects died, the most commonly recorded cause of death being cardiovascular disease. Duration adjusted odds ratios (OR) and 95% confidence intervals (CI) were calculated separately for Type 1 and Type 2 subjects. The mortality rates for men were higher than for women (Type 1: OR 1.27, CI 0.61-2.62; Type 2: OR 1.79, CI 1.27-2.52); were higher for those of lower vs higher social class (Type 1: OR 1.34, CI 0.61-2.96; Type 2: OR 2.0, CI 1.41-2.85); and were higher for those who left school before 16 years of age compared to those who left school at or after 16 years of age (Type 1: OR 3.98, CI 1.96-8.06; Type 2: OR 2.86, CI 1.93-4.25). Subjects who were unemployed had a higher mortality rate than those employed at the time of the study (Type 1: OR 3.10, CI 1.67-5.79; Type 2: OR 2.88, CI 2.12-3.91) and those living in council housing had a greater mortality than those who were living in other types of housing (Type 1: OR 2.57, CI 1.35-4.91, Type 2: OR 2.76, CI 2.05-3.73). Also for both Type 1 and Type 2 subjects mortality was significantly higher in those subjects who had a least one diabetic complication at baseline and reported one or more hospital admissions in the previous year and in Type 2 subjects with poor glycaemic control. After adjusting for duration of diabetes, hospital admissions, and the presence of diabetic complications, being unemployed, male, in poor glycaemic control (Type 2 only), and less educated were significant risk factors for mortality (p<0.001). These results suggest that there are important indicators of social deprivation which predict mortality over and above diabetic health status itself. Locally targeted action will be required if these inequalities in health experienced by people with diabetes are to be reduced.


Anticoagulation for people with cancer and central venous catheters.
Central venous catheter (CVC) placement increases the risk of thrombosis in people with cancer. Thrombosis often necessitates the removal of the CVC, resulting in treatment delays and thrombosis-related morbidity and mortality. To evaluate the relative efficacy and safety of anticoagulation for thromboprophylaxis in people with cancer with a CVC. We searched the Cochrane Central Register of Controlled Trials (CENTRAL, Issue 12, 2012), MEDLINE Ovid (January 1966 to February 2013), and EMBASE Ovid (1980 to February 2013). We handsearched conference proceedings, checked references of included studies, used the 'related citations' feature within PubMed, and searched clinicaltrials.gov for ongoing studies. Randomized controlled trials (RCTs) comparing the effects of any dose of unfractionated heparin (UFH), low-molecular-weight heparin (LMWH), vitamin K antagonists (VKA), or fondaparinux with no intervention or placebo or comparing the effects of two different anticoagulants in people with cancer and a CVC. Teams of two review authors independently used a standardized form to extract data in duplicate. They resolved any disagreements by discussion. They extracted data on risk of bias, participants, interventions, and outcomes. Outcomes of interest included mortality, symptomatic deep venous thrombosis (DVT), asymptomatic DVT, major bleeding, minor bleeding, infection, and thrombocytopenia. Where possible, we conducted meta-analyses using the random-effects model. Of 9559 identified citations, we included 12 RCTs (17 publications) reporting follow-up data on 2823 participants. Two of the RCTs included children. Of the 10 RCTs including 2564 adults, one compared prophylactic dose heparin with low-dose VKA. Three RCTs compared VKA with no VKA and four RCTs compared heparin with no heparin. Two additional trials had three separate arms comparing heparin, VKA, and no intervention. Prophylactic-dose heparin, compared with no heparin, was associated with a statistically significant reduction in symptomatic DVT (risk ratio (RR) 0.48; 95% confidence interval (CI) 0.27 to 0.86; moderate-quality evidence). However, results did not confirm or exclude a beneficial or detrimental effect of heparin on mortality (RR 0.82; 95% CI 0.53 to 1.26; moderate-quality evidence), major bleeding (RR 0.49; 95% CI 0.03 to 7.84; low-quality evidence), infection (RR 1.00; 95% CI 0.54 to 1.85; moderate-quality evidence); thrombocytopenia (RR 1.03; 95% CI 0.80 to 1.33; moderate-quality evidence), or minor bleeding (RR 1.35; 95% CI: 0.62 to 2.92). Low-dose VKAs, compared with no VKAs, were associated with a statistically significant reduction in asymptomatic DVT (RR 0.43; 95% CI 0.30 to 0.62). Results did not confirm or exclude a beneficial or detrimental effect of VKAs on mortality (RR 1.04; 95% CI 0.89 to 1.22; low-quality evidence), symptomatic DVT (RR 0.51; 95% CI 0.21 to 1.22; low-quality evidence), major bleeding (RR 7.60; 95% CI 0.94 to 61.49; very-low-quality evidence), or minor bleeding (RR 3.14; 95% CI 0.14 to 71.51). The use of heparin, compared with VKA was associated with a statistically significant increase in thrombocytopenia (RR 3.73; 95% CI 2.26 to 6.16; low-quality evidence) and asymptomatic DVT (RR 1.74; 95% CI 1.20 to 2.52). However, results did not show or exclude a beneficial or detrimental effect on any of the other outcomes of interest (very-low-quality evidence). Compared with no anticoagulation, we found a statistically significant reduction of symptomatic DVT with heparin and asymptomatic DVT with VKA. Heparin was associated with a higher risk of thrombocytopenia and asymptomatic DVT when compared with VKA. However, the findings did not rule out other clinically important benefits and harms. People with cancer with CVCs considering anticoagulation should balance the possible benefit of reduced thromboembolic complications with the possible harms and burden of anticoagulants.


Chromosomal phylogeny of Robertsonian races of the house mouse on the island of Madeira: testing between alternative mutational processes.
The ancestral karyotype of the house mouse (Mus musculus) consists of 40 acrocentric chromosomes, but numerous races exist within the domesticus subspecies characterized by different metacentric chromosomes formed by the joining at the centromere of two acrocentrics. An exemplary case is present on the island of Madeira where six highly divergent chromosomal races have accumulated different combinations of 20 metacentrics in 500-1000 years. Chromosomal cladistic phylogenies were performed to test the relative performance of Robertsonian (Rb) fusions, Rb fissions and whole-arm reciprocal translocations (WARTs) in resolving relationships between the chromosomal races. The different trees yielded roughly similar topologies, but varied in the number of steps and branch support. The analyses using Rb fusions/fissions as characters resulted in poorly supported trees requiring six to eight homoplasious events. Allowance for WARTs considerably increased nodal support and yielded the most parsimonious trees since homoplasy was reduced to a single event. The WART-based trees required five to nine WARTs and 12 to 16 Rb fusions. These analyses provide support for the role of WARTs in generating the extensive chromosomal diversification observed in house mice. The repeated occurrence of Rb fusions and WARTs highlights the contribution of centromere-related rearrangements to accelerated rates of chromosomal change in the house mouse.


Nonrandom Distribution of Azole Resistance across the Global Population of Aspergillus fumigatus.
The emergence of azole resistance in the pathogenic fungus Aspergillus fumigatus has continued to increase, with the dominant resistance mechanisms, consisting of a 34-nucleotide tandem repeat (TR34)/L98H and TR46/Y121F/T289A, now showing a structured global distribution. Using hierarchical clustering and multivariate analysis of 4,049 A. fumigatus isolates collected worldwide and genotyped at nine microsatellite loci using analysis of short tandem repeats of A. fumigatus (STRAf), we show that A. fumigatus can be subdivided into two broad clades and that cyp51A alleles TR34/L98H and TR46/Y121F/T289A are unevenly distributed across these two populations. Diversity indices show that azole-resistant isolates are genetically depauperate compared to their wild-type counterparts, compatible with selective sweeps accompanying the selection of beneficial mutations. Strikingly, we found that azole-resistant clones with identical microsatellite profiles were globally distributed and sourced from both clinical and environmental locations, confirming that azole resistance is an international public health concern. Our work provides a framework for the analysis of A. fumigatus isolates based on their microsatellite profile, which we have incorporated into a freely available, user-friendly R Shiny application (AfumID) that provides clinicians and researchers with a method for the fast, automated characterization of A. fumigatus genetic relatedness. Our study highlights the effect that azole drug resistance is having on the genetic diversity of A. fumigatus and emphasizes its global importance upon this medically important pathogenic fungus.IMPORTANCE Azole drug resistance in the human-pathogenic fungus Aspergillus fumigatus continues to emerge, potentially leading to untreatable aspergillosis in immunosuppressed hosts. Two dominant, environmentally associated resistance mechanisms, which are thought to have evolved through selection by the agricultural application of azole fungicides, are now distributed globally. Understanding the effect that azole resistance is having on the genetic diversity and global population of A. fumigatus will help mitigate drug-resistant aspergillosis and maintain the azole class of fungicides for future use in both medicine and crop protection.


Focusing light inside live tissue using reversibly switchable bacterial phytochrome as a genetically encoded photochromic guide star.
Focusing light deep by engineering wavefronts toward guide stars inside scattering media has potential biomedical applications in imaging, manipulation, stimulation, and therapy. However, the lack of endogenous guide stars in biological tissue hinders its translations to in vivo applications. Here, we use a reversibly switchable bacterial phytochrome protein as a genetically encoded photochromic guide star (GePGS) in living tissue to tag photons at targeted locations, achieving light focusing inside the tissue by wavefront shaping. As bacterial phytochrome-based GePGS absorbs light differently upon far-red and near-infrared illumination, a large dynamic absorption contrast can be created to tag photons inside tissue. By modulating the GePGS at a distinctive frequency, we suppressed the competition between GePGS and tissue motions and formed tight foci inside mouse tumors in vivo and acute mouse brain tissue, thus improving light delivery efficiency and specificity. Spectral multiplexing of GePGS proteins with different colors is an attractive possibility.


Review of radiation issues for computed tomography.
Over the past three decades, computed tomography (CT) has proven to be central in imaging evaluation. Multidetector technology continues to drive practice patterns by combining fast scanning with high quality data sets. This has resulted in new applications as well as improved use in traditional applications. With this recognition has also come the realization that there are potential costs of CT. One major cost is the radiation dose. Therefore, in order to begin to assess benefits (which are relatively familiar to radiologists) versus costs (which are less familiar), the issues related to CT radiation need to be addressed. Familiarity with measures of CT radiation and the actual doses delivered by CT are important issues as they provide a basis for understanding the potential cancer risks from CT radiation. Moreover, these justify development of strategies to minimize radiation dose. Strategies include obtaining only necessary CT examinations and adjusting the examinations based on scan indication, region examined, and patient size. These strategies must also be combined with efforts by manufacturers in development and implementation of technology aimed at radiation dose management, as well as efforts in research, education, and CT standards and regulation. This article reviews the subject of radiation dose with multidetector CT technology, including measures of CT radiation, the dose that can result from CT examinations, the risks of this amount of radiation, and strategies for minimizing CT radiation dose.


siRNA-directed DNA Methylation in Plants.
DNA cytosine methylationis an important epigenetic process that is correlated with transgene silencing, transposon suppression, and gene imprinting. In plants, small interfering RNAs (siRNAs) can trigger DNA methylation at loci containing their homolog sequences through a process called RNA-directed DNA methylation (RdDM). In canonical RdDM, 24 nucleotide (nt) siRNAs (ra-siRNAs) will be loaded into their effector protein called ARGONAUTE 4 (AGO4) and subsequently targeted to RdDM loci through base-pairing with the non-coding transcripts produced by DNA-directed RNA Polymerase V. Then, the AGO4-ra-siRNA will recruit the DNA methyltransferase to catalyze de novo DNA methylation. Recent studies also identified non-canonical RdDM pathways that involve microRNAs or 21 nt siRNAs. These RdDM pathways are biologically important since they control responses biotic and abiotic stresses, maintain genome stability and regulate development. Here, we summarize recent pro-gresses of mechanisms governing canonical and non-canonical RdDM pathways.


900 MHz electromagnetic field exposure affects qualitative and quantitative features of hippocampal pyramidal cells in the adult female rat.
The effects of electromagnetic fields (EMFs) emitted by mobile phones on humans hold special interest due to their use in close proximity to the brain. The current study investigated the number of pyramidal cells in the cornu ammonis (CA) of the 16-week-old female rat hippocampus following postnatal exposure to a 900 megahertz (MHz) EMF. In this study were three groups of 6 rats: control (Cont), sham exposed (Sham), and EMF exposed (EMF). EMF group rats were exposed to 900 MHz EMF (1 h/day for 28 days) in an exposure tube. Sham group was placed in the exposure tube but not exposed to EMF (1 h/day for 28 days). Cont group was not placed into the exposure tube nor were they exposed to EMF during the study period. In EMF group rats, the specific energy absorption rate (SAR) varied between 0.016 (whole body) and 2 W/kg (locally in the head). All of the rats were sacrificed at the end of the experiment and the number of pyramidal cells in the CA was estimated using the optical fractionator technique. Histopathological evaluations were made on sections of the CA region of the hippocampus. Results showed that postnatal EMF exposure caused a significant decrease of the pyramidal cell number in the CA of the EMF group (P<0.05). Additionally, cell loss can be seen in the CA region of EMF group even at qualitative observation. These results may encourage researchers to evaluate the chronic effects of 900 MHz EMF on teenagers' brains.


Determinants of management errors in acute severe asthma.
It is hypothesised that, despite recent initiatives to improve asthma self-management including asthma education, detailed investigation of the sequence of events culminating in hospital admission will lead to the identification of important management errors and thus the likelihood that the majority of severe asthma attacks are preventable by currently available strategies, and that psychological, health care and socioeconomic factors are risk factors for such management errors. A cross sectional study was undertaken of 138 patients aged 15-50 years admitted to hospital (general ward or intensive care unit) with acute severe asthma who were assessed within 24-72 hours of admission using a number of previously validated instruments. A detailed history of events of the attack was assessed against predetermined criteria for non or delayed use of oral corticosteroids and non or delayed use of emergency ambulance services. Subjects had evidence of severe chronic asthma and had acute severe asthma at presentation (n = 90, pH = 7.3 (0.2), PaCO2 = 7.2 (5.0) kPa) but duration of hospital stay was short (3.7 (2.6) days). Serious management errors occurred very frequently and most were deemed to have been made by the patient. Forward stepwise regression revealed that delayed or non-use of oral corticosteroids was predicted independently by lack of paying job (p = 0.02), high total use of inhaled beta agonists in the 24 hours before index admission (p = 0.04), loss of a job in the last year (p = 0.04), low frequency of use of oral corticosteroids in the last year (p = 0.06), concerns during the index attack about medical expenses (p = 0.07), and delay in the use of ambulance services (p = 0.05)--the model being responsible for 23% of the variance. Delayed or non-summoning of emergency ambulance services was predicted independently by total life events (p = 0.03), having something stolen in the last year (p = 0.003), panic during the index attack (p = 0.01), and concerns during the index attack about taking time off work (p = 0.07)--the model being responsible for 21% of the variance. The results of this study show that, despite recent educational advances, serious management errors are common in those admitted to hospital with acute severe asthma and that most management errors relate to patient self-management behaviour. Serious management errors are predicted by a variety of socio-economic and psychological factors. While the results of this study are consistent with the widely held view that most acute severe attacks are theoretically preventable, the challenge for the future is to change patients' behaviour in the face of considerable adverse socioeconomic and psychological factors.


MYB Labeling by Immunohistochemistry Is More Sensitive and Specific for Breast Adenoid Cystic Carcinoma than MYB Labeling by FISH.
Breast adenoid cystic carcinoma (ACC) is a primary breast carcinoma that, like salivary gland ACC, displays the t(6;9) translocation resulting in the MYB-NFIB gene fusion and immunopositivity for MYB by immunohistochemistry (IHC). However, it is not well established whether MYB immunoreactivity or rearrangement can be used to support a diagnosis of ACC in a malignant basaloid or benign cribriform breast lesion. Whole sections of primary breast ACC (n=11), collagenous spherulosis (CS; n=7), and microglandular adenosis (MGA; n=5) and tissue microarrays containing 16 basal-like, triple-negative breast carcinomas (TNBC) were labeled for MYB by IHC and underwent MYB fluorescence in situ hybridization using a break-apart probe. Strong, diffuse nuclear MYB labeling was seen in 100% ACC compared with no cases of basal-like TNBC, CS, or MGA (P=0.0001). Any degree of nuclear MYB labeling was seen in 100% ACC compared with 54% of all other cases (P=0.007), with any labeling seen in 71% CS, 63% basal-like TNBC, and 0% MGA. MYB rearrangement was detected in 89% (8/9) of evaluable ACC compared with 4% (1/26) of all other evaluable cases (P=0.0001), with a rearrangement detected in 1 (7%; n=1/15) evaluable basal-like TNBC. Strong, diffuse nuclear labeling for MYB is more sensitive than MYB fluorescence in situ hybridization for breast ACC and can be used to support a diagnosis of ACC in a cribriform or basaloid lesion in the breast. However, weak and focal labeling should be interpreted with caution as it can be seen in other benign cribriform and malignant basaloid lesions.


Breast cancer screening, incidence, and mortality in West Virginia.
Situated entirely in the Appalachian region, West Virginia (WV) as a state is characterized by a diverse population, with a major portion of the population residing in rural areas. Breast cancer is the most commonly diagnosed cancer among women residents of the state. Using different data sources, we have reported the most recent screening, incidence, and mortality information associated with breast cancer in WV. In 2006, roughly one-fourth of women > or =40 years of age had not had a mammogram in the past two years. Sociodemographic factors were found to be associated with lack of mammography screening among these women. The leveling off of the increase in the in-situ breast cancer incidence rates in WV since 1999 is closely associated with the bottoming out of the proportion of women without mammography that has been seen since early this decade. Counties in southern and northern part of the state generally have higher invasive breast cancer incidence rates than those in the central part. Breast cancer-related mortality is higher among southern counties in comparison to counties in northern and central part of the state, with mortality rates higher than the national average in McDowell, Mingo, Wayne, and Mason Counties. There is a compelling need to increase access to mammography screening among women residents in the state.