Evaluation of upper airway characteristics in patients with and without sleep apnea using cone-beam computed tomography and computational fluid dynamics.
To determine if upper airway characteristics and airway pressure change significantly between low risk, healthy non-OSA subjects, and OSA subjects during respiration using cone-beam computed tomography (CBCT) imaging and steady-state k-ω model computational fluid dynamics (CFD) fluid flow simulations, respectively. CBCT scans were collected at both end-inhalation and end-exhalation for 16 low-risk non-OSA subjects and compared to existing CBCT data from 7 OSA subjects. The CBCT images were imported into Dolphin Imaging and the upper airway was segmented into stereolithography (STL) files for area and volumetric measurements. Subject models that met pre-processing criteria underwent CFD simulations using ANSYS Fluent Meshing (Canonsburg, PA) in which unstructured mesh models were generated to solve the standard dual equation turbulence model (k-ω). Objective and supplemental descriptive measures were obtained and statistical analyses were performed with both parametric and non-parametric tests to evaluate statistical significance at P < .05. Regarding area and volumetric assessments, there were statistically significant mean differences in Total Volume and Minimum CSA between non-OSA and OSA groups at inhalation and exhalation (P = .002, .003, .004, and .007), respectively. There were also statistically significant mean differences in volume and min CSA between the inhalation and exhalation for the non-OSA group (P < .001 and .002), respectively. While analysis of the CFD simulation was limited by the collected data available, a finding consistent with published literature was that the OSA subject group simulation models depicted the point of lowest pressure coinciding with the area of maximum constriction.


Silencing of ribonucleotide reductase subunit M1 potentiates the antitumor activity of gemcitabine in resistant cancer cells.
Gemcitabine is a deoxycytidine analog used for the treatment of a wide range of solid tumors. Its efficacy is however often reduced due to the development of resistance. Ribonucleotide reductase M1 subunit (RRM1) is a key determinant of gemcitabine resistance, and tumor cells that overexpress RRM1 are resistant to the cytotoxicity of gemcitabine. In the present study, we showed that RRM1-specific small interfering RNA (siRNA), when complexed with polyethylenimine, effectively downregulated the expression of RRM1 protein in mouse tumor cells that overexpress RRM1, both in vitro and in vivo. More importantly, systemic administration of the RRM1-specific siRNA significantly inhibited the growth of RRM1-overexpressing tumors in mice and sensitized the tumors to gemcitabine treatment. These findings suggest that silencing RRM1 expression using siRNA could potentially be an effective strategy to overcome gemcitabine resistance.


Normative performance of older individuals on the Hopkins Verbal Learning Test-Revised (HVLT-R) according to ethno-racial group, gender, age and education level.
The Hopkins Verbal Learning Test-Revised (HVLT-R) provides a measure of verbal learning and memory. The aim of this study was to provide normative performance data on the HVLT-R for community-dwelling older individuals according to ethno-racial group, age, gender, and years of completed education, in Australia and the U.S. The ASPirin in Reducing Events in the Elderly (ASPREE) study recruited 19,114 generally healthy community dwelling individuals aged 70 years and over (65 years and over for U.S minorities), who were without a diagnosis of dementia and scored above 77 on the modified Mini-Mental State (3MS) examination. Included in the analysis presented here were 16,251 white Australians, and in the U.S. 1,082 white, 894 African American and 314 Hispanic/Latino individuals at baseline. Performance on each of the components of the HVLT-R (trials 1-3, total, learning, delayed recall, delayed recognition, percentage retention and recognition discrimination index [RDI]) differed by demographic variables. In country and ethno-racial stratified analyses, female gender, younger age and higher education were significantly associated with better total recall, delayed recall and RDI. Among white Australians these characteristics were also associated with better retention. Age, education and gender-specific reference values across ethno-racial categories were determined. Ethno-racial, age, gender and education-stratified normative data from this large cohort of community-dwelling older individuals will serve as important reference standards in Australia and the U.S. to assess cognition in older individuals.


Long-Term Endocrine Outcomes Following Endoscopic Endonasal Transsphenoidal Surgery for Acromegaly and Associated Prognostic Factors.
Long-term remission rates from endoscopic transsphenoidal surgery for acromegaly and their relationship to prognostic indicators of disease aggressiveness are not well documented. To investigate long-term remission rates in patients with acromegaly after endoscopic transsphenoidal surgery, and correlate this with molecular and radiographic markers of disease aggressiveness. We identified all patients undergoing endoscopic transsphenoidal surgery for acromegaly from 2005 to 2013 at Cedars-Sinai Pituitary Center. Hormonal remission was established by normal insulin-like growth factor (IGF)-1, basal serum growth hormone <2.5 ng/mL, and growth hormone suppression to <1 ng/mL following oral glucose tolerance test. Oral glucose tolerance test was performed at 3 months after surgery, and then as indicated. IGF-1 was measured at 3 months and then at least annually. We evaluated tumor granularity, nuclear expression of p21, Ki67 index, and extent of cavernous sinus invasion, and correlated these with remission status. Fifty-eight patients that underwent surgery had follow-up from 38 to 98 months (mean 64 ± 32.2 months). There were 21 microadenomas and 37 macroadenomas. Three months after surgery 40 of 58 patients (69%) were in biochemical remission. Four additional patients were in remission at 6 months after surgery, and 1 patient had recurrence within the first year after surgery. At last follow-up, 43 of 44 (74.1%) of patients remained in remission. Cavernous sinus invasion by tumor predicted failure to achieve remission. Prognostic markers of disease aggressiveness other than cavernous sinus invasion did not correlate with surgical outcome. Long-term remission after surgery alone was achieved in 74% of patients, indicating long-term efficacy of endoscopic surgery.


Risk of chronic arthropathy among women after rubella vaccination. Vaccine Safety Datalink Team.
A review by the Institute of Medicine found a possible relationship between rubella vaccination and chronic arthritis among women. To evaluate the risk of persistent joint and neurologic symptoms in rubella seronegative women subsequently vaccinated with RA 27/3 rubella vaccine. Retrospective cohort study based on computerized laboratory data and medical record review. Records were reviewed for symptoms occurring within 2 years before and after the date of serological testing and to identify vaccinees. Possible cases were evaluated by a rheumatologist blinded to serological findings and vaccination status. Large health maintenance organization in northern California. Women aged 15 to 59 years serotested for rubella during 1990 with continuous health plan membership for 2 years before and after the date of their serological test. Seronegative women immunized within 1 year of serotesting (n=971) were defined as exposed. Primary comparison groups included all unvaccinated, seronegative women (n=924) and randomly selected seropositive, unvaccinated women (n=2421) matched to exposed subjects on serological test date and age (+/-3 years). Prevalence and incidence of chronic joint and neurologic symptoms during 1-year follow-up period stratified by age and serological findings, immunization, and postpartum status. No significantly increased risk was associated with receipt of rubella vaccine for any outcome except for prevalence of carpal tunnel syndrome in vaccinated women at least 30 years old compared with seropositive, unvaccinated women (2.9% vs 1.4%; P=.03). A total of 34 women had onset of conditions within the 1-year follow-up period; 9 of these were in the group of seronegative, immunized women, of whom 6 had onset of symptoms within 6 weeks of vaccination. Among these 6 women, symptoms included transient arthritis or arthralgias (<6 weeks duration) in 4 women, arthralgia of indeterminate chronicity in 1 woman, and carpal tunnel syndrome in 1 woman. Postpartum women across all groups were less likely to be seen for nontraumatic arthropathies than nonpostpartum women (4.5% vs 7.2%, P=.08 in vaccinated women; 4.8% vs 8.1%, P=.09 in seronegative controls; and 4.8% vs 10.0%, P=.01 in seropositive controls). In this large retrospective cohort analysis there was no evidence of any increased risk of new onset chronic arthropathies or neurologic conditions in women receiving the RA 27/3 rubella vaccine. These data support the continued vaccination of rubella-susceptible women to reduce the risk of congenital rubella syndrome.


Toluene concentrations in the blood and alveolar air of workers during the workshift and the morning after.
Occupational toluene exposure was studied during the workshift and the morning after by the analysis of environmental air, alveolar air, and blood. Environmental toluene exposure was measured by both continuous and instantaneous sampling. Instantaneous environmental toluene concentrations correlated better with alveolar toluene concentrations (r = 0.94; n = 155) than with blood toluene concentrations (r = 0.71; n = 52). Continuous environmental toluene concentrations correlated better with blood toluene concentrations (r = 0.84; n = 65) than with alveolar toluene concentrations (r = 0.52; n = 46). During the workshift and the morning after, blood and alveolar toluene concentrations correlated significantly with each other (r = 0.75; n = 66 and r = 0.67; n = 52). In a group of workers who were exposed to a mean environmental toluene concentration of 146 micromilligrams the concentrations of toluene in the alveolar air and blood the morning after were 3.2 micromilligrams (SD = 1.7) and 27.5 micromilligrams (SD = 12.7) respectively. With regard to the morning after toluene determinations, blood concentrations correlated (r = 0.52; n = 52; p less than 0.001) better than the alveolar concentrations with the corresponding afternoon values (r = 0.36; n = 52; p less than 0.01). The decline of the toluene concentrations from the end of one workshift to the start of the next exposure indicated a mean toluene half life of 3.8 hours in the alveolar air and of 4.5 hours in blood and therefore the 17 hour interval between two consecutive workshifts was insufficient for the complete elimination of absorbed toluene.


Viral infection triggers interferon-induced expulsion of live Cryptococcus neoformans by macrophages.
Cryptococcus neoformans is an opportunistic human pathogen, which causes serious disease in immunocompromised hosts. Infection with this pathogen is particularly relevant in HIV+ patients, where it leads to around 200,000 deaths per annum. A key feature of cryptococcal pathogenesis is the ability of the fungus to survive and replicate within the phagosome of macrophages, as well as its ability to be expelled from host cells via a novel non-lytic mechanism known as vomocytosis. Here we show that cryptococcal vomocytosis from macrophages is strongly enhanced by viral coinfection, without altering phagocytosis or intracellular proliferation of the fungus. This effect occurs with distinct, unrelated human viral pathogens and is recapitulated when macrophages are stimulated with the anti-viral cytokines interferon alpha or beta (IFNα or IFNβ). Importantly, the effect is abrogated when type-I interferon signalling is blocked, thus underscoring the importance of type-I interferons in this phenomenon. Lastly, our data help resolve previous, contradictory animal studies on the impact of type I interferons on cryptococcal pathogenesis and suggest that secondary viral stimuli may alter patterns of cryptococcal dissemination in the host.


Anatomical and electrical remodeling with incomplete left atrial appendage ligation: Results from the LAALA-AF registry.
The anatomical, electrical, and clinical impact of incomplete Lariat left atrial appendage ligation remains unclear. We studied LAA anatomy pre- and postligation using contrast enhanced-computed tomography (CT) scans in 91 patients with atrial fibrillation (AF) who subsequently underwent catheter ablation (CA). Eleven patients had an incomplete exclusion (12%) with a central leak ranging from 1 to 5 mm. Despite incomplete ligation; the LAA volume were reduced by 67% postprocedurally when compared to preprocedure. In 7 patients with a leak between 1 and 3 mm, there was a 77% reduction in LAA volume beyond the ligation site suggestive of remodeling of the LAA. In 4 patients with larger (4-5 mm) leak the LAA remnants (LAARs) were slightly larger than those with smaller leaks on follow-up CT scan. Three out of the 4 demonstrated LAA electrical activity during CA and underwent isolation of the LAA ostium. Follow-up imaging showed two of these LAARs completely sealed with no communication with the left atrium. There was no significant difference in the AF recurrence rates between the patients who had a leak versus those with complete ligation (4 of 11 [36%] vs. 22 of 80 [27%]; P = 0.6). Oral anticoagulation was discontinued in all patients with small leaks and 2 patients with large leaks that sealed completely upon follow-up imaging. There were no strokes or TIAs at 12 months. Despite incomplete LAA ligation by Lariat device there is significant anatomical and electrical remodeling that resulted in reduction in LAA size, volume, and electrical activity.


Genetic structure of bluefin tuna in the mediterranean sea correlates with environmental variables.
Atlantic Bluefin Tuna (ABFT) shows complex demography and ecological variation in the Mediterranean Sea. Genetic surveys have detected significant, although weak, signals of population structuring; catch series analyses and tagging programs identified complex ABFT spatial dynamics and migration patterns. Here, we tested the hypothesis that the genetic structure of the ABFT in the Mediterranean is correlated with mean surface temperature and salinity. We used six samples collected from Western and Central Mediterranean integrated with a new sample collected from the recently identified easternmost reproductive area of Levantine Sea. To assess population structure in the Mediterranean we used a multidisciplinary framework combining classical population genetics, spatial and Bayesian clustering methods and a multivariate approach based on factor analysis. FST analysis and Bayesian clustering methods detected several subpopulations in the Mediterranean, a result also supported by multivariate analyses. In addition, we identified significant correlations of genetic diversity with mean salinity and surface temperature values revealing that ABFT is genetically structured along two environmental gradients. These results suggest that a preference for some spawning habitat conditions could contribute to shape ABFT genetic structuring in the Mediterranean. However, further studies should be performed to assess to what extent ABFT spawning behaviour in the Mediterranean Sea can be affected by environmental variation.


Assessment of Diastolic Function and Thiol-Disulphide Homeostasis in Arsenic-Exposed Workers.
Exposure to arsenic is associated with various cardiovascular diseases. The imbalance between antioxidant and oxidant homeostasis plays a crucial role in the cardiovascular effects of arsenic. The aim of this study was to investigate the effect of arsenic exposure on diastolic function by measuring thiol and disulphide in arsenic-exposed workers. A total of 107 male arsenic-exposed workers and 36 healthy subjects were enrolled. Mitral inflow velocity and parameters of diastolic function were measured. As oxidative stress indicators, total thiol, native thiol, disulphide, and their percent ratios were determined. The mean age was 39.1 ± 9.5 years in the arsenic-exposed group and 37.4 ± 9.6 years in the controls. The median blood arsenic level was 42 μg/dL in the arsenic-exposed group and 3.75 μg/dL in the controls. E-wave, E/A ratio, and e' wave were lower and left atrial diameter, A-wave, average E/e' ratio, and tricuspid regurgitation velocity were higher in the arsenic-exposed group. Native and total thiol concentrations were lower, and disulphide/native and disulphide/total thiol ratios were higher in the arsenic-exposed group. Fourteen (13.1%) workers had diastolic dysfunction, 26 (24.3%) had indeterminate, and 67 (62.6%) had normal diastolic function, compared to 1 (2.8%), 2 (5.6%), and 33 (97.7%) in the control group, respectively. In regression analysis, disulphide/native thiol ratio (p < 0.001) and blood arsenic level (p < 0.001) predicted increased average E/e' ratio in the arsenic-exposed group. This study showed strong associations among arsenic exposure, oxidative stress, and diastolic function, and revealed the influence of arsenic exposure on diastolic dysfunction through oxidative stress.


Tocopherol efficacy and safety for preventing retinopathy of prematurity: a randomized, controlled, double-masked trial.
To test the efficacy and safety of vitamin E in preventing retinopathy of prematurity, 287 infants with birth weights of less than 1.5 kg or gestational ages of less than 33 weeks were enrolled within 24 hours of birth in a randomized, double-masked trial of IV, followed by oral, placebo v tocopherol (adjusted to plasma levels of 3 to 3.5 mg/dL). In the 196 infants completing ophthalmic follow-up, tocopherol did not prevent retinopathy of prematurity of any stage (28% placebo treated v 26% tocopherol treated) or moderately severe retinopathy of prematurity (8% placebo treated v 11% tocopherol treated). Cicatricial sequelae were not significantly different (1/97 placebo treated v 3/99 tocopherol treated), with one placebo-treated infant and one tocopherol-treated infant having retinal detachments. Among all 232 infants examined, those treated with tocopherol had more retinal hemorrhage than placebo-treated infants (8/121 placebo treated v 16/111 tocopherol treated), and retinal hemorrhage correlated positively (P less than .01) with plasma levels of tocopherol after the first 2 weeks of age. Prospective monitoring of morbidity including late-onset sepsis, necrotizing enterocolitis, etc revealed no differences between groups except that grades 3 and 4 intraventricular hemorrhage occurred more frequently in infants weighing less than 1 kg at birth who had received tocopherol (14/42, 33%) v those who had received placebo (4/43, 9%) (P less than .02). Our data do not support the use of tocopherol for prophylaxis against retinopathy of prematurity in premature infants and suggest that IV tocopherol treatment starting on day 1 may increase the incidence of hemorrhagic complications of prematurity, particularly in infants with birth weights of less than 1 kg.


O-prenylated carbostyrils as a novel class of 15-lipoxygenase inhibitors: Synthesis, characterization, and inhibitory assessment.
Catalyzed peroxidation of unsaturated lipid in animals and plants intimately is linked to the activity of 15-Lipoxygenase enzymes. Lipoxygenases (LOXs) are well known to play an important role in many acute and chronic syndromes such as inflammation, asthma, cancer, and allergy. In this study, a series of mono prenyloxycarbostyrils were synthesized and evaluated as potential inhibitors of soybean 15-Lipoxygenase (SLO) and their inhibitory potencies were compared to mono prenyloxycoumarins which had been reported in the previous works. The synthetic compounds inhibit lipoxygenase enzyme by competitive mechanism like the prenyloxy coumarins. The results showed that position and length of the prenyl moiety play the important role in lipoxygenase inhibitory activity. Among all of the synthetic compounds (coumarin and carbostyril derivatives), 5-farnesyloxycoumarin and 8-farnesyloxycarbostyril demonstrated the best inhibitory activity by IC50 values of 1.1 µM and 0.53 µM, respectively.


The neuropathology of obesity: insights from human disease.
Obesity, a pathologic state defined by excess adipose tissue, is a significant public health problem as it affects a large proportion of individuals and is linked with increased risk for numerous chronic diseases. Obesity is the result of fundamental changes associated with modern society including overnutrition and sedentary lifestyles. Proper energy homeostasis is dependent on normal brain function as the master metabolic regulator, which integrates peripheral signals, modulates autonomic outflow and controls feeding behavior. Therefore, many human brain diseases are associated with obesity. This review explores the neuropathology of obesity by examining brain diseases which either cause or are influenced by obesity. First, several genetic and acquired brain diseases are discussed as a means to understand the central regulation of peripheral metabolism. These diseases range from monogenetic causes of obesity (leptin deficiency, MC4R deficiency, Bardet-Biedl syndrome and others) to complex neurodevelopmental disorders (Prader-Willi syndrome and Sim1 deficiency) and neurodegenerative conditions (frontotemporal dementia and Gourmand's syndrome) and serve to highlight the central regulatory mechanisms which have evolved to maintain energy homeostasis. Next, to examine the effect of obesity on the brain, chronic neuropathologic conditions (epilepsy, multiple sclerosis and Alzheimer's disease) are discussed as examples of obesity leading to maladaptive processes which exacerbate chronic disease. Thus, obesity is associated with multiple pathways including abnormal metabolism, altered hormonal signaling and increased inflammation which act in concert to promote downstream neuropathology. Finally, the effect of anti-obesity interventions is discussed in terms of brain structure and function. Together, understanding human diseases and anti-obesity interventions leads to insights into the bidirectional interaction between peripheral metabolism and central brain function, highlighting the need for continued clinicopathologic and mechanistic studies of the neuropathology of obesity.


Lactobacillus reuteri extracts promoted wound healing via PI3K/AKT/β-catenin/TGFβ1 pathway.
The balance of oral microbiomes is crucial to maintain oral health. Microecological imbalance can impair the function of mesenchymal stem cells (MSCs) and lead to delay wound healing. Probiotics is a promising prevention approach for the treatment of oral inflammatory diseases caused by a bacterial infection. However, the effect of probiotics on oral MSCs and wound healing is unclear. In the present study, we used one type of probiotics Lactobacillus reuteri extracts to determine whether bacterial extracts could regulate the functions of gingiva MSCs (GMSCs) and promote wound healing. Lactobacillus reuteri was prepared with bacterial extracts using ultrasonic crushing apparatus. The effects of Lactobacillus reuteri extracts on GMSCs were tested using the cell scratch migration, alkaline phosphatase (ALP) activity, alizarin red staining, cell counting kit-8, real-time PCR, and western blot assays. To investigate the role of Lactobacillus reuteri extracts in the wound in mice, the wound position of bilateral mesial gingival of the maxillary first molar was established, the wound area with a size of 1 mm × 2 mm and the full thickness gingiva was removed. Mice with wound were randomly distributed to two groups: injection of 0.9% NaCl (NS group) or injection of 50 μg/ml bacterial extracts. We discovered that 50 μg/ml Lactobacillus reuteri extracts increased the capacities of migration, expression of stem cell markers, osteogenic differentiation, and proliferation of GMSCs. In addition, local injection of 50 μg/ml bacterial extracts could promote wound-healing process in mice models. Mechanistically, we found that Lactobacillus reuteri extracts accelerated the process of wound healing via PI3K/AKT/β-catenin/TGFβ1 pathway. These data showed that Lactobacillus reuteri extracts could activate the potentials of GMSCs, thus promote wound healing. Our discovery provided the insight of the underlying mechanism activating functions of MSCs and identified Lactobacillus reuteri extracts as a potential therapeutic strategy for accelerating oral wound and potential application in the future dental clinic.


Phylogenetic and biogeographic implications inferred by mitochondrial intergenic region analyses and ITS1-5.8S-ITS2 of the entomopathogenic fungi Beauveria bassiana and B. brongniartii.
The entomopathogenic fungi of the genus Beauveria are cosmopolitan with a variety of different insect hosts. The two most important species, B. bassiana and B. brongniartii, have already been used as biological control agents of pests in agriculture and as models for the study of insect host - pathogen interactions. Mitochondrial (mt) genomes, due to their properties to evolve faster than the nuclear DNA, to contain introns and mobile elements and to exhibit extended polymorphisms, are ideal tools to examine genetic diversity within fungal populations and genetically identify a species or a particular isolate. Moreover, mt intergenic region can provide valuable phylogenetic information to study the biogeography of the fungus. The complete mt genomes of B. bassiana (32,263 bp) and B. brongniartii (33,920 bp) were fully analysed. Apart from a typical gene content and organization, the Beauveria mt genomes contained several introns and had longer intergenic regions when compared with their close relatives. The phylogenetic diversity of a population of 84 Beauveria strains -mainly B. bassiana (n = 76) - isolated from temperate, sub-tropical and tropical habitats was examined by analyzing the nucleotide sequences of two mt intergenic regions (atp6-rns and nad3-atp9) and the nuclear ITS1-5.8S-ITS2 domain. Mt sequences allowed better differentiation of strains than the ITS region. Based on mt and the concatenated dataset of all genes, the B. bassiana strains were placed into two main clades: (a) the B. bassiana s. l. and (b) the "pseudobassiana". The combination of molecular phylogeny with criteria of geographic and climatic origin showed for the first time in entomopathogenic fungi, that the B. bassiana s. l. can be subdivided into seven clusters with common climate characteristics. This study indicates that mt genomes and in particular intergenic regions provide molecular phylogeny tools that combined with criteria of geographic and climatic origin can subdivide the B. bassiana s.l. entomopathogenic fungi into seven clusters with common climate characteristics.


The extracellular matrix molecule tenascin-C modulates cell cycle progression and motility of adult neural stem/progenitor cells from the subependymal zone.
Adult neurogenesis has been described in two canonical regions of the adult central nervous system (CNS) of rodents, the subgranular zone (SGZ) of the hippocampus and the subependymal zone (SEZ) of the lateral ventricles. The stem cell niche of the SEZ provides a privileged environment composed of a specialized extracellular matrix (ECM) that comprises the glycoproteins tenascin-C (Tnc) and laminin-1 (LN1). In the present study, we investigated the function of these ECM glycoproteins in the adult stem cell niche. Adult neural stem/progenitor cells (aNSPCs) of the SEZ were prepared from wild type (Tnc+/+) and Tnc knockout (Tnc-/-) mice and analyzed using molecular and cell biological approaches. A delayed maturation of aNSPCs in Tnc-/- tissue was reflected by a reduced capacity to form neurospheres in response to epidermal growth factor (EGF). To examine a potential influence of the ECM on cell proliferation, aNSPCs of both genotypes were studied by cell tracking using digital video microscopy. aNSPCs were cultivated on three different substrates, namely, poly-D-lysine (PDL) and PDL replenished with either LN1 or Tnc for up to 6 days in vitro. On each of the three substrates aNSPCs displayed lineage trees that could be investigated with regard to cell cycle length. The latter appeared reduced in Tnc-/- aNSPCs on PDL and LN1 substrates, less so on Tnc that seemed to compensate the absence of the ECM compound to some extent. Close inspection of the lineage trees revealed a subpopulation of late dividing aNSPCslate that engaged into cycling after a notable delay. aNSPCslate exhibited a clearly different morphology, with a larger cell body and conspicuous processes. aNSPCslate reiterated the reduction in cell cycle length on all substrates tested, which was not rescued on Tnc substrates. When the migratory activity of aNSPC-derived progeny was determined, Tnc-/- neuroblasts displayed significantly longer migration tracks. This was traced to an increased rate of migration episodes compared to the wild-type cells that rested for longer time periods. We conclude that Tnc intervenes in the proliferation of aNSPCs and modulates the motility of neuroblasts in the niche of the SEZ.


'Early' mammalian myoblasts are resistant to phorbol ester-induced block of differentiation.
Mesenchymal cells were isolated from somites and limbs of mouse embryos at different developmental stages. When grown in tissue culture, some of the cells underwent muscle differentiation as indicated by synthesis of sarcomeric myosin, acetylcholine receptor and, in the case of limb cells, fusion into multinucleated myotubes. When the tumour promoter 12-O-tetradecanoyl phorbol 13-acetate (TPA) was added to these cultures, it caused differential effects, depending upon the age of the embryo from which cells were isolated. In cultures of somites or limb bud from embryos up to 12 days post coitum, TPA did not interfere with the appearance of differentiated muscle cells. When TPA was added to cultures from older embryos, it inhibited muscle differentiation with an efficiency which increased with the age of the embryo, reaching about 90% inhibition at 15 days. After this period, a new population of myogenic cells appeared in the limb, which were able to differentiate in the presence of TPA and represented the great majority of myoblasts after day 18 of embryonic development. The simplest interpretation of these data can be based on the existence of three major classes of myogenic cell precursors, which appear sequentially during muscle histogenesis: 'early' myoblasts, which appear resistant to tumour promoters; 'late' myoblasts, whose differentiation is inhibited by tumour promoters and 'satellite' cells which, like early myoblasts, show no sensitivity to TPA.


Evaluation of motion correction methods in human brain PET imaging--a simulation study based on human motion data.
Motion correction in PET has become more important as system resolution has improved. The purpose of this study was to evaluate the accuracy of event-by-event and frame-based MC methods in human brain PET imaging. Motion compensated image reconstructions were performed with static and dynamic simulated high resolution research tomograph data with frame-based image reconstructions, using a range of measured human head motion data. Image intensities in high-contrast regions of interest (ROI) and parameter estimates in tracer kinetic models were assessed to evaluate the accuracy of the motion correction methods. Given accurate motion data, event-by-event motion correction can reliably correct for head motions. The average ROI intensities and the kinetic parameter estimates VT and BPND were comparable to the true values. The frame-based motion correction methods with correctly aligned attenuation map using the average of externally acquired motion data or motion data derived from image registration give comparable quantitative accuracy. For large intraframe (>5 mm) motion, the frame-based methods produced ≈ 9% bias in ROI intensities, ≈ 5% in VT, and ≈ 10% in BPND estimates. In addition, in real studies that lack a ground truth, the normalized weighted residual sum of squared difference is a potential figure-of-merit to evaluate the accuracy of motion correction methods. The authors conclude that frame-based motion correction methods are accurate when the intraframe motion is less than 5 mm and when the attenuation map is accurately aligned. Given accurate motion data, event-by-event motion correction can reliably correct for head motion in human brain PET studies.


The application of genetic knowledge: ethical and policy implications.
This article explores societal ethical concerns about the application of the rapidly developing knowledge in the field of genetics. The article calls for nurses to reflect on these ethical concerns not only in the care of patients but also in professional and societal discussion, so that sound ethical and societally acceptable public policy can be adopted. A brief summary introduces the concepts of ethics and policy. The text than offers discussion of genetic testing for adults and children, testing in relation to behavioral attributes, and genetics and reproduction. Hypothetical cases using fictional names are used to illustrate each section. Recommendations for nurse action are cited. Finally, the article poses questions that require public awareness and discussion.


Microsomal N-oxidation of the hepatocarcinogen N-methyl-4-aminoazobenzene and the reactivity of N-hydroxy-N-methyl-4-aminoazobenzene.
The N-oxidation of the hepatocarcinogen N-methyl-4-aminoazobenzene (MAB) was catalyzed by hepatic microsomes in a reduced pyridine nucleotide- and oxygen-dependent reaction. The initial N-oxidation product, N-hydroxy-N-methyl-4-aminoazobenzene (N-HO-MAB), was readily oxidized to a second product that yielded N-hydroxy-4-aminoazobenzene upon subsequent acid treatment. The secondary N-oxidation product may be formed nonenzymatically and is presumed to be N-HO-MAB N-oxide or its dehydrated derivative, N-(p-phenylazophenyl)nitrone. Under the same conditions, MAB was also oxidatively N-dealkylated to 4-aminoazobenzene, which was N-oxidized to N-hydroxy-4-aminoazobenzene. Unlike the latter reactions, the microsomal N-oxidation of MAB was independent of cytochrome P-450, as shown by its lack of sensitivity to inhibition by 2-[(2,4-dichloro-6-phenyl)phenoxy]ethylamine and its inability to utilize cumene hydroperoxide in place of reduced pyridine nucleotides and oxygen. The N-oxidation of MAB was also catalyzed by the purified microsomal flavoprotein mixed-function amine oxidase of Ziegler et al. The noncarcinogenic dye N-ethyl-4-aminoazobenzene was metabolized similarly to MAB. For male animals the hepatic levels of MAB N-oxidase activity were in the order: rat greater than hamster, guinea pig greater than mouse, rabbit. Little or no MAB N-oxidase activity was present in several extrahepatic rat tissues. N-HO-MAB, N-hydroxy-N-ethyl-4-aminoazobenzene, and N-hydroxy-4-aminoazobenzene catalyzed the aerobic oxidation of cysteine and glutathione. These hydroxylamines also bound covalently to proteins. The binding of N-HO-MAB with nucleic acids was only 3 to 6% that observed with serum albumin. Under anhydrous conditions the nitrone generated aerobically from N-HO-MAB reacted with carbon-carbon or carbon-nitrogen double bonds, or both, in fatty acids, retinol, purines, and pyrimidines to yield isoxazolidine and/or oxadiazolidine addition products. The nitrone from N-hydroxy-N-ethyl-4-aminoazobenzene was much less reactive under these conditions. Syntheses of N-HO-MAB and N-hydroxy-N-ethyl-4-aminoazobenzene are reported.


Glucocorticoid repression of the mouse gonadotropin-releasing hormone gene is mediated by promoter elements that are recognized by heteromeric complexes containing glucocorticoid receptor.
We have identified two regions of the mouse gonadotropin-releasing hormone (GnRH) promoter, one between -237 and -201 (distal element) and the other between -184 and -150 (proximal element), which are required for glucocorticoid repression in transiently transfected GT1-7 cells. These sequences show no similarity to known positive or negative glucocorticoid response elements (nGREs) and do not function when placed upstream of heterologous viral promoters. The glucocorticoid receptor (GR) does not bind directly to the distal or proximal promoter elements but may participate in glucocorticoid repression of GnRH gene transcription by virtue of its association within multiprotein complexes at these nGREs. Electrophoretic mobility shift assays with GT1-7 nuclear extract demonstrate the presence of GR-containing protein complexes on GnRH nGREs. One protein that co-occupies the distal nGRE in vitro along with GR is the POU domain transcription factor Oct-1. Thus, the tethering of GR to the GnRH distal nGRE, by virtue of a direct or indirect association with DNA-bound Oct-1, could play a role in hormone-dependent transcriptional repression of the GnRH gene. In contrast, Oct-1 does not appear to be a component of the GR-containing protein complex that is bound to the proximal nGRE.


Onset of the release of spermatozoa (spermarche) in boys in relation to age, testicular growth, pubic hair, and height.
The onset of production of spermatozoa (spermarche) is the basis for achievement of reproductive capacity in men. We collected 24-h urine samples every 3 months in a 7-yr longitudinal study of 40 normal boys initially aged 8.6-11.7 yr. After centrifugation, the urine was analyzed for the presence of spermatozoa by microscopic examination, and spermarche was estimated on the basis of age at first observed spermaturia. The results were corrected for the intermittent occurrence of spermatozoa in the urine after first observed spermaturia and the fact that the urine samples were collected quarterly. In addition, physical examination, including determination of testicular size by orchidometer measurement, pubic hair distribution (Tanner stage), and height, was carried out every 6 months. Spermarche occurred at a median age of 13.4 yr (range, 11.7-15.3 yr), at a time when testicular size was 4.7-19.6 ml (median, 11.5 ml), and pubic hair distribution was 1-5 (median, 2.5). In most boys, spermarche preceded the age of peak height velocity (median, 13.8 yr; range, 12.2-15.2 yr); at the time of spermarche, median peak height growth velocity was 9.9 cm/yr (range, 7.5-13.4 cm/yr), and median height was 160.4 cm (range, 151.7-175.9 cm). We conclude that spermarche is an early pubertal event and that a wide variation in testicular size and secondary sex characteristics is found at that time. In particular, spermarche may occur when little or no pubic hair has developed, and the testes have grown only slightly.


Epidemiology of mandibular fractures in a tertiary trauma centre.
Fractures of the mandible are common facial injuries. Patients frequently require hospitalisation, surgical intervention and extended periods of convalescence. A prospective database of patients presenting to the Oral and Maxillofacial Surgery service at Christchurch Hospital during an 11-year period was reviewed. 1045 patients with mandibular fractures were identified. Variables examined included demographic data, type of fractures, mode of injury and treatment delivered. More than 90% of patients were men, with 64% in the 15-29 years age group. Interpersonal violence accounted for 49% of fractures, followed by sports (16%), falls (13%) and motor vehicle accidents (10%). The condyle was the most frequent fracture site (34%) and multiple fractures were seen in 37% of patients. Hospitalisation was required for 53% of patients with 89% of these treatments being open reduction and internal fixation. Mandibular fracture is a common facial injury. The incidence is highest in young men who are victims of interpersonal violence. Alcohol is a major contributing factor. Management involved hospitalisation and surgical intervention for more than half of those presenting.


Transcriptomics analysis of differentially expressed genes in subcutaneous and perirenal adipose tissue of sheep as affected by their pre- and early postnatal malnutrition histories.
Early life malnutrition is known to target adipose tissue with varying impact depending on timing of the insult. This study aimed to identify differentially expressed genes in subcutaneous (SUB) and perirenal (PER) adipose tissue of 2.5-years old sheep to elucidate the biology underlying differential impacts of late gestation versus early postnatal malnutrition on functional development of adipose tissues. Adipose tissues were obtained from 37 adult sheep born as twins to dams fed either NORM (fulfilling energy and protein requirements), LOW (50% of NORM) or HIGH (110% of protein and 150% of energy requirements) diets in the last 6-weeks of gestation. From day 3 to 6 months of age, lambs were fed high-carbohydrate-high-fat (HCHF) or moderate low-fat (CONV) diets, and thereafter the same moderate low-fat diet. The gene expression profile of SUB in the adult sheep was not affected by the pre- or early postnatal nutrition history. In PER, 993 and 186 differentially expressed genes (DEGs) were identified in LOW versus HIGH and NORM, respectively, but no DEG was found between HIGH and NORM. DEGs identified in the mismatched pre- and postnatal nutrition groups LOW-HCHF (101) and HIGH-HCHF (192) were largely downregulated compared to NORM-CONV. Out of 831 DEGs, 595 and 236 were up- and downregulated in HCHF versus CONV, respectively. The functional enrichment analyses revealed that transmembrane (ion) transport activities, motor activities related to cytoskeletal and spermatozoa function (microtubules and the cytoskeletal motor protein, dynein), and responsiveness to the (micro) environmental extracellular conditions, including endocrine and nervous stimuli were enriched in the DEGs of LOW versus HIGH and NORM. We confirmed that mismatched pre- and postnatal feeding was associated with long-term programming of adipose tissue remodeling and immunity-related pathways. In agreement with phenotypic measurements, early postnatal HCHF feeding targeted pathways involved in kidney cell differentiation, and mismatched LOW-HCHF sheep had specific impairments in cholesterol metabolism pathways. Both pre- and postnatal malnutrition differentially programmed (patho-) physiological pathways with implications for adipose functional development associated with metabolic dysfunctions, and PER was a major target.


[Basic components in the development of research proposals in health sciences].
Any investigation in the health sciences should have a plan or proposal to answer a specific research question. This document, designed for persons preparing for or already pursuing a research project, describes the conceptual framework related with a research proposal, from the study design until its implementation. The objective of this document is to summarize different theoretical aspects that should be considered in a research project, including pertinent references that might be used to deepen the discussion. First, a series of definitions related to the scientific method are presented and then the processes to carry out a research proposal including common epidemiologic designs are illustrated. In addition, a series of research questions that can serve as a guide to perform a critical analysis of published papers are included. Finally, an interdisciplinary group integrated by health professionals with the clinical expertise and health professionals with the necessary skills in biostatistics and epidemiology to undertake a sound scientific research is recommended.


EASL Clinical Practice Guideline: Occupational liver diseases.
A variety of chemicals have been linked to occupational liver diseases, including several solvents and mixtures thereof, pesticides, and metals. Workplace exposures have been associated with virtually the entire spectrum of acute and chronic liver diseases. However, their prevalence is inadequately quantified and their epidemiology limited. Occupational liver diseases may result from high accidental or from prolonged lower level exposures. Whereas the former is uncommon and easily recognised, the latter are relatively more frequent but often overlooked because they may display normal values of conventional markers, have an insidious onset and be asymptomatic or be obfuscated and confounded by concurrent conditions. In addition, specific tests of toxicity are not available, histopathology may not be revealing and the assessment of internal dose of chemicals is usually not decisive. Given these circumstances, the diagnosis of these liver disorders is challenging, one of exclusion and often requires an interdisciplinary approach. These recommendations offer a classification of the type of liver injuries associated with occupational exposures - based in part on the criteria for drug-induced liver injury - a grading of their severity, and the diagnostic and preventive criteria for chemically induced occupational liver disease.


Early effects of mannitol in patients with head injuries assessed using bedside multimodality monitoring.
We have employed bedside multimodality methods to assess the influence of a slow (20 min) bolus of hypertonic mannitol on cerebral hemodynamics in comatose patients with head injuries. Middle cerebral artery flow velocities (FV) and cortical microcirculatory flows were measured in comatose patients with head injuries after the administration of 200 ml of 20% mannitol. A comparison was made with the effects of an identical bolus of isotonic saline. Fourteen patients with diffuse head injuries and with raised intracranial pressure were selected, and mannitol infusion studies were conducted when clinically indicated (n = 23). Using transcranial doppler and laser doppler flowmetry (LDF), indices of estimated cerebrovascular resistance (eCVR) were calculated for the macro- (eCVR-FV) and micro- (eCVR-LDF) circulation. During mannitol infusion, a significant rise in cerebral perfusion pressure was detected (+10%, P = 0.03) as a result of a fall in intracranial pressure (-21%, P = 0.001). Increases in both FV (+13%, P < 0.001) and LDF (+14%, P = 0.002) occurred only after the administration of mannitol and persisted beyond completion of infusion. The effect on FV and LDF decayed exponentially, with a time constant of 34.0 and 38.0 minutes, respectively, and was independent of the pressure autoregulatory status. There was a tendency for eCVR-FV and eCVR-LDF to decrease. No significant effects resulted from the administration of saline. Bedside multimodality monitoring may provide a useful means for assessing the effects of therapy in the comatose patient. The mechanisms by which mannitol reduces intracranial pressure in patients with head injuries are discussed.


Isolation of mutations in the alpha operon of Escherichia coli that suppress the transcriptional defect conferred by a mutation in the porin regulatory gene envZ.
One class of mutations in the envZ gene of Escherichia coli K-12 confers a pleiotropic defect on the expression of several genes, including ompF, lamB, and phoA, that are otherwise not commonly regulated. Four second-site mutations that suppress this transcriptional defect have been isolated by using a procedure that circumvented the problem of intragenic suppressors, including true revertants. All four mutations have been mapped to the genes of the alpha operon and have been assigned tentatively to the gene rpoA, which specifies the alpha subunit of RNA polymerase. The mutations, referred to as sez (for suppressor of envZ), did not appear to confer a phenotype on an otherwise wild-type strain and did not suppress the transcriptional defects conferred by several other phenotypic classes of envZ mutations, including amber mutations. Our results led us to postulate that the alpha subunit or some other component of the alpha operon plays a role in determining the specificity of gene expression.


Use of on-line UV/Vis-spectrometry in the measurement of dissolved ozone and AOC concentrations in drinking water treatment.
The concentrations of dissolved ozone and assimilable organic carbon (AOC) are important performance parameters in drinking water production. For the measurement of ozone, a spectral algorithm was developed that allows quantification in situ using a UV/Vis spectrometer probe. Furthermore, a strong correlation between the change in the absorption spectrum after individual treatment steps and the formation or removal of AOC in that treatment step was observed. This allowed the development of a spectral algorithm that predicts AOC formation during ozonation and subsequent removal in further treatment steps. This method has been verified at one pilot plant of the Amsterdam drinking water supply.


Robust prostate cancer marker genes emerge from direct integration of inter-study microarray data.
DNA microarray data analysis has been used previously to identify marker genes which discriminate cancer from normal samples. However, due to the limited sample size of each study, there are few common markers among different studies of the same cancer. With the rapid accumulation of microarray data, it is of great interest to integrate inter-study microarray data to increase sample size, which could lead to the discovery of more reliable markers. We present a novel, simple method of integrating different microarray datasets to identify marker genes and apply the method to prostate cancer datasets. In this study, by applying a new statistical method, referred to as the top-scoring pair (TSP) classifier, we have identified a pair of robust marker genes (HPN and STAT6) by integrating microarray datasets from three different prostate cancer studies. Cross-platform validation shows that the TSP classifier built from the marker gene pair, which simply compares relative expression values, achieves high accuracy, sensitivity and specificity on independent datasets generated using various array platforms. Our findings suggest a new model for the discovery of marker genes from accumulated microarray data and demonstrate how the great wealth of microarray data can be exploited to increase the power of statistical analysis. email@example.com.


Compact dipole nanoantenna coupler to plasmonic slot waveguide.
Optical nanoantennas can be used for coupling radiation to or from waveguides in analogy to micro- and radio-wave systems. In this letter we provide a systematic description of the design approaches for a coupler to a plasmonic slot waveguide in the telecom range around 1.55 µm with realistic excitation from a lensed optical fiber. We show that the best coupling efficiency of 26% can be achieved by utilizing a dipole antenna with side and bottom reflectors, and such coupling efficiency is 185 times larger than for the bare waveguide. The nanoantenna coupler provides a compact interface between an optical fiber and a plasmonic slot waveguide for future optical integrated circuits.


Methylergonovine maleate as a cluster headache prophylactic: a study and review.
Methylergonovine maleate (Methergine), an ergot derivative with vasoconstrictive properties, has been cited as an effective treatment for vascular headaches. Few studies are available to support its use in headache management. An uncontrolled pilot study of 20 episodic cluster headache patients confirmed its effectiveness and tolerability as an adjunct cluster headache prophylactic. Decreased headache frequency was reported by 19 of 20 patients (95%), and 15 of 20 patients (75%) reported decreased intensity of headaches within 1 week of initiating therapy. A review of methylergonovine's pharmacokinetic, molecular, and tolerability profile clarifies its mechanisms and clinical role in headache management.


Structural analysis of a periplasmic binding protein in the tripartite ATP-independent transporter family reveals a tetrameric assembly that may have a role in ligand transport.
Several bacterial solute transport mechanisms involve members of the periplasmic binding protein (PBP) superfamily that bind and deliver ligand to integral membrane transport proteins in the ATP-binding cassette, tripartite tricarboxylate transporter, or tripartite ATP-independent (TRAP) families. PBPs involved in ATP-binding cassette transport systems have been well characterized, but only a few PBPs involved in TRAP transport have been studied. We have measured the thermal stability, determined the oligomerization state by small angle x-ray scattering, and solved the x-ray crystal structure to 1.9 A resolution of a TRAP-PBP (open reading frame tm0322) from the hyperthermophilic bacterium Thermotoga maritima (TM0322). The overall fold of TM0322 is similar to other TRAP transport related PBPs, although the structural similarity of backbone atoms (2.5-3.1 A root mean square deviation) is unusually low for PBPs within the same group. Individual monomers within the tetrameric asymmetric unit of TM0322 exhibit high root mean square deviation (0.9 A) to each other as a consequence of conformational heterogeneity in their binding pockets. The gel filtration elution profile and the small angle x-ray scattering analysis indicate that TM0322 assembles as dimers in solution that in turn assemble into a dimer of dimers in the crystallographic asymmetric unit. Tetramerization has been previously observed in another TRAP-PBP (the Rhodobacter sphaeroides alpha-keto acid-binding protein) where quaternary structure formation is postulated to be an important requisite for the transmembrane transport process.


Establishing Trust When Assessing Learners: Barriers and Opportunities.
Trust plays a critical role in the assessment of learners in the clinical setting. In an ideal system, learners can be vulnerable and share their limitations and areas for improvement, while faculty possess the time and skill to provide specific feedback that enables learners to achieve competency in clinical skills. For medical students, a number of threats to the establishment of trust in the learning environment exist, including the interplay between feedback and grades, the existence of bias, and competing demands for faculty time. However, several strategies can help institutions to overcome these threats and foster a culture of trust related to assessment and assessment systems: Provide ungraded environments where learners are able to be vulnerable, cocreate assessments and assessment systems with faculty and learners, acknowledge and address bias, and provide faculty with adequate time and resources to employ best practices in assessment. By intentionally employing these strategies, our institutions can support trust in assessment systems and further learner growth and achievement.


Retaining nurses: the impact of Ontario's "70% full-time commitment".
To evaluate the effectiveness of a nurse retention strategy, the "70% Full-Time Commitment", in retaining part-time and casual nurses in Ontario's nurse profession. Using the College of Nurses of Ontario database, a longitudinal dataset for all nurses registered with the college from 1993 to 2006 was created (N=216,353). One-year transition probabilities of nurse employment status (full-time, part-time and casual) were conducted (1993-2009) to generate trends of nurses' likelihood to stay in, switch or leave their full-time, part-time or casual position in Ontario's nurse profession. After the 70% Full-Time Commitment (2004-2009) was initiated, most full-time (89.7%), part-time (76.6%) and casual (62.5%) nurses stayed in their employment position. A slightly larger proportion of part-time nurses (13.6%) switched to full-time compared to casual nurses (8.6%). However, a similar proportion of young part-time (24.5%) and casual (23.3%) nurses switched to full-time. A smaller proportion of part-time (3.2%) and casual (7.1%) nurses left the profession. Part-time and casual nurses have different employment switching patterns. The "70% Full-Time Commitment" was not an effective mechanism in retaining part-time and casual nurses. It might be more effective as a recruitment initiative for young nurses.


[Basic cardiopulmonary resuscitation].
Since the introduction of the modern techniques for cardiopulmonary resuscitation in the 1960s, professionals have discussed the need to standardize its application and teaching both among health care professionals and the general public. The "ILCOR", Committee to Coordinate Resuscitation Techniques, in an effort to simplify resuscitation techniques, set out some recommendations in August 2000 which were adopted by the leading organizations such as AHA and ERC, in charge of diffusing Vital Suport techniques. In these, different levels of attention have been incorporated depending on the qualifications which the person has who is provide this treatment. The most important changes are the necessity to put Emergency Medical Services into action rapidly; the techniques to follow if there are one or two persons apply resuscitation techniques; for artificial resuscition emergency care, new volumes depending on the use or non-use of oxygen; the acceptance of devices to open up alternate air passageways; recommendations whether or not it is pertinent to check on a patient's pulse depending on the qualifications of the person attending that patient; heart message techniques exterpulmonary resuscition woth only thorax compression and automatic external defibrillation.


The effects of kindling on GABA-mediated inhibition in the dentate gyrus of the rat. I. Paired-pulse depression.
Double-pulse stimulation of the perforant path input to the dentate gyrus was used in the following experiments to produce paired pulse depression in that site. This effect provided an estimate of GABA-mediated recurrent inhibition. The depression was enhanced by drugs that facilitate or mimic GABA action and attenuated by drugs which block GABA transmission. Paired-pulse depression was significantly increased following amygdala kindling and was further enhanced to near maximal levels by subsequent kindling in the dentate. In addition, kindling did not increase the rate at which inhibition failed under conditions of excessive activation. Trains of 5 Hz stimulation, applied to the perforant path, caused paired-pulse depression to disappear and elicited a brief AD. Following kindling, the latency to AD onset tended to be increased rather than shortened, suggesting an enhanced resistance to inhibitory failure. These results indicated that kindling increased, rather than reduced, inhibition in the dentate gyrus.


The Doctorate in Clinical Laboratory Sciences: A New Curriculum to Enhance the Connection of the Laboratory to Health Care Providers.
This report discusses the need for a Doctorate in Clinical Laboratory Sciences program and describes a curriculum to train Doctorate in Clinical Laboratory Sciences students. The Doctorate in Clinical Laboratory Sciences program was developed to help reduce diagnostic errors in patient care by enhancing connections between the clinical laboratory and health care providers. Data are presented from program implementation in 2016 to 2017 academic year to 2019 to 2020 regarding the faculty and student demographics, program statistics (eg, admissions and attrition rates), and effectiveness. Perceptions of program effectiveness were obtained via surveys from 28 faculty physicians who supervised Doctorate in Clinical Laboratory Sciences students during clinical service rotations. Another survey assessed the preferred type of practice after graduation of 33 students. Over the 4-year period, the program had a 50% rate of admission and a 21.8% attrition rate. As of December 2020, 15 students graduated from the program. The majority (69%-82%) of physician faculty who completed the survey agreed that Doctorate in Clinical Laboratory Sciences students contributed positively at clinical rounds. Approximately two-thirds of students reported a preference to lead a Diagnostic Management Team or serve as an advanced practice provider in a Diagnostic Management Team with leadership provided by an MD/DO or PhD. This report provides useful information for other institutions that may want to establish similar Doctorate in Clinical Laboratory Sciences programs. Early data suggest that our program effectively trains doctoral-level advanced practice medical laboratory scientists, who may play an important role in improving patient safety by reducing diagnostic errors and providing value-based, optimal patient care.


C2'-pyrene-functionalized triazole-linked DNA: universal DNA/RNA hybridization probes.
Development of universal hybridization probes, that is, oligonucleotides displaying identical affinity toward matched and mismatched DNA/RNA targets, has been a longstanding goal due to potential applications as degenerate PCR primers and microarray probes. The classic approach toward this end has been the use of "universal bases" that either are based on hydrogen-bonding purine derivatives or aromatic base analogues without hydrogen-bonding capabilities. However, development of probes that result in truly universal hybridization without compromising duplex thermostability has proven challenging. Here we have used the "click reaction" to synthesize four C2'-pyrene-functionalized triazole-linked 2'-deoxyuridine phosphoramidites. We demonstrate that oligodeoxyribonucleotides modified with the corresponding monomers display (a) minimally decreased thermal affinity toward DNA/RNA complements relative to reference strands, (b) highly robust universal hybridization characteristics (average differences in thermal denaturation temperatures of matched vs mismatched duplexes involving monomer W are <1.7 °C), and (c) exceptional affinity toward DNA targets containing abasic sites opposite of the modification site (ΔT(m) up to +25 °C). The latter observation, along with results from absorption and fluorescence spectroscopy, suggests that the pyrene moiety is intercalating into the duplex whereby the opposing nucleotide is pushed into an extrahelical position. These properties render C2'-pyrene-functionalized triazole-linked DNA as promising universal hybridization probes for applications in nucleic acid chemistry and biotechnology.


The hereditary autoinflammatory disorders uncovered.
There is a thriving interest in the field of hereditary autoinflammatory disorders (HAID), a gamut of heterogeneous conditions deriving from an aberrant orchestration of innate immunity, unified by the common feature of seemingly unprovoked inflammation, which might be systemic or occur in localized niches of the organism. Recurrent fever and episodic inflammation in the joints, serosal membranes, skin, gut, and other organs are the common denominator of HAID. Mutations in the inflammasome-related genes have been associated with different HAID, showing the intimate link existing between interleukin-1 (IL-1)-structured inflammasome and their pathogenesis. Differential diagnosis of HAID can be challenging, as there are no universally accepted diagnostic protocols, and near half of patients may remain without any genetic abnormality identified. The use of IL-1-antagonists has been associated with beneficial effects in a large number of HAID associated with excessive IL-1 signalling, such as cryopyrin-associated periodic syndromes, familial Mediterranean fever, and deficiency of IL-1 receptor antagonist. This review will discuss about the key-clues of HAID which might guide for an early recognition and drive decisions for treatment.


Cerebral oedema and increased intracranial pressure in chronic liver disease.
Cerebral oedema is a cause of morbidity and mortality in fulminant hepatic failure but has not been well documented as a complication of chronic liver diseases. We report here the development of cerebral oedema and increased intracranial pressure in 12 patients with chronic liver disease. Between July 1, 1987, and Dec 31, 1993, we studied 12 patients aged 29-67 years with end-stage chronic liver disease. All the patients had cirrhosis, portal hypertension, hypoprothrombinaemia, hepatic encephalopathy, and decreased serum concentrations of albumin (<25 g/L). During the study, the patients developed signs of increased intracranial pressure and had documented intracranial hypertension, cerebral oedema, or both. Intracranial hypertension was suspected on physical examination and confirmed by epidural catheters. We detected cerebral oedema by computed axial tomography of the head and necropsy of the brain when possible. All the patients had intracranial hypertension and cerebral oedema. Two patients had successful treatment of cerebral hypertension with improvement of intracranial pressure such that orthotopic liver transplantation was undertaken. Both patients became neurologically normal after transplantation. Eight patients had only a transient response to treatment and died of cerebral oedema before a transplant could be done. Cerebral oedema and increased intracranial pressure can occur in chronic liver disease and presents as neurological deterioration. Treatment guided by monitoring of intracranial pressure can lead to the reversal of intracranial hypertension, but in most patients cerebral oedema contributes to death or places them at too high a risk for liver transplantation.


Reverse lactate threshold: a novel single-session approach to reliable high-resolution estimation of the anaerobic threshold.
The multisession maximal lactate steady-state (MLSS) test is the gold standard for anaerobic threshold (AnT) estimation. However, it is highly impractical, requires high fitness level, and suffers additional shortcomings. Existing single-session AnT-estimating tests are of compromised validity, reliability, and resolution. The presented reverse lactate threshold test (RLT) is a single-session, AnT-estimating test, aimed at avoiding the pitfalls of existing tests. It is based on the novel concept of identifying blood lactate's maximal appearance-disappearance equilibrium by approaching the AnT from higher, rather than from lower exercise intensities. Rowing, cycling, and running case data (4 recreational and competitive athletes, male and female, aged 17-39 y) are presented. Subjects performed the RLT test and, on a separate session, a single 30-min MLSS-type verification test at the RLT-determined intensity. The RLT and its MLSS verification exhibited exceptional agreement at 0.5% discrepancy or better. The RLT's training sensitivity was demonstrated by a case of 2.5-mo training regimen following which the RLT's 15-W improvement was fully MLSS-verified. The RLT's test-retest reliability was examined in 10 trained and untrained subjects. Test 2 differed from test 1 by only 0.3% with an intraclass correlation of 0.997. The data suggest RLT to accurately and reliably estimate AnT (as represented by MLSS verification) with high resolution and in distinctly different sports and to be sensitive to training adaptations. Compared with MLSS, the single-session RLT is highly practical and its lower fitness requirements make it applicable to athletes and untrained individuals alike. Further research is needed to establish RLT's validity and accuracy in larger samples.


Transfer of student learning in medical education.
Transfer of learning among the cognitive, psychomotor, and affective domains and among three clinical disciplines--medicine, pediatrics, and surgery--was examined in the final year of a medical student clerkship program. Rather than following the traditional psychological approach, the authors used a model based on ethnographic analysis followed by performance measurement. The general assumption that what students learn in one subject area is transferred to another area was questioned. A total of 120 medical students were assessed in knowledge, skills, and attitudes prior to and after three clinical rotations. The results indicated that learning was discipline-specific as well as specific to the learning domain studied. Therefore, no generalizations can be made from the study of one subject area or one specific aspect of student learning to another. The use of an alternative methodology to study complex behavior rather than the traditional paradigm is suggested.


Developmental and environmental factors that enhance binding of Bordetella pertussis to human epithelial cells in relation to sudden infant death syndrome (SIDS).
Asymptomatic infection due to Bordetella pertussis has been suggested to be one cause of sudden infant death syndrome (SIDS). We examined developmental and environmental factors previously found to affect binding of another toxigenic species, Staphylococcus aureus, to human epithelial cells: expression of the Lewis(a) antigen; infection with respiratory syncytial virus (RSV); exposure to cigarette smoke; and the inhibitory effect of breast milk on bacterial binding. Binding of two strains of B. pertussis (8002 and 250825) to buccal epithelial cells was significantly reduced by treating the cells with monoclonal antibodies to Lewis(a) (P < 0.05) and Lewis(x) (P < 0.01) antigens. Both strains bound in significantly greater numbers to cells from smokers compared with cells from non-smokers (P < 0.05). HEp-2 cells infected with RSV subtypes A or B had higher binding indices for both 8002 (P < 0.001) and 250825 (P < 0.01). On RSV-infected cells, there was significantly enhanced binding of monoclonal antibodies to Lewis(x) (P < 0.05), CD14 (P < 0.001) and CD18 (P < 0.01); and pre-treatment of cells with anti-CD14 or CD18 also significantly reduced binding of both strains of B. pertussis. Pre-treatment of the bacteria with human milk significantly reduced their binding to epithelial cells. The results are discussed in relation to our three-year survey of bacterial carriage among 253 healthy infants, their mothers and local SIDS cases between 1993-1995 and in relation to the change to an earlier immunisation schedule for infants and the recent decline in SIDS in Britain.


Physician response to a medication alert system in inpatients with levodopa-treated diseases.
To evaluate the appropriateness of dopamine receptor antagonist prescriptions in hospitalized patients with dopamine-requiring diseases after implementation of an automated prescription alert system. We examined dopamine receptor antagonist prescriptions in hospitalized patients with dopamine-requiring diseases and physician response to an automated drug contraindication alert system at Barnes-Jewish Hospital from 2009 to 2013. A detailed review of patient medical records was performed for all alert events generated when a physician prescribed a dopamine receptor antagonist concurrently with a dopamine receptor agonist in hospitalized patients. Two movement disorders neurologists determined the appropriateness of each prescription, based on patient medical history, through consensus. Physician response to alert was compared by indication for the prescription and physician specialty. Of 237 orders, 197 (83.1%) prescriptions for dopamine receptor antagonists were considered inappropriate. The prevalence of inappropriate dopamine receptor antagonist prescriptions per levodopa prescriptions was 16.10% (95% confidence interval 9.47, 22.73) in psychiatry, 7.51% (6.16, 8.86) in general medicine, 6.14% (4.49, 7.79) in the surgical specialties, and 0.85% (0.46, 1.25) in the neurologic/neurosurgical specialties. Of the inappropriate prescriptions, 146 (74.1%) were continued despite the alert. The strongest predictor of discontinuation of dopamine receptor antagonist medications was use of the medication to treat nausea or emesis (p < 0.001). Despite successfully identifying instances when dopamine antagonists were prescribed to patients with dopamine-requiring diseases, the alert system modestly affected physician prescribing behavior, highlighting the need for improved education of health care providers.


Comparison of gene expression profiles induced by fresh or ozone-oxidized black carbon particles in A549 cells.
Epidemiological studies have showed an association between black carbon (BC) exposure and adverse health effects. This study intends to investigate the influence of oxidation processes in atmosphere on the initial cellular responses of BC. The changes of gene expressions induced by fresh BC (FBC) and ozone-oxidized BC (OBC) in human lung epithelial A549 cells were analyzed. And their toxic effects presented by viability, LDH release and DNA damage were compared. Totally 47, 000 genes in A549 cells were examined using Affymetrix Human U133 plus 2.0 chips. Some of the differentially expressed genes were verified by reverse transcription-quantitative polymerase chain reaction (RT-qPCR). The results showed that 1446 genes (including 756 up-regulated and 690 down-regulated) and 1594 genes (including 788 up-regulated and 806 down-regulated genes) were significantly changed by FBC and OBC respectively. Only 4 of 14 (FBC)/15 (OBC) oxidative stress related genes, up- or down-regulated by FBC and OBC, were identical; 13 of 29 (FBC)/31 (OBC) inflammation related genes, and 6 of 20 (FBC)/18 (OBC) autophagy related genes were identical. No obvious differences were observed between the toxic effects of FBC and OBC. The cytotoxicity of OBC and FBC in A549 cells is at least partially induced by oxidative stress and consequent inflammation or autophagy process. Previous studies indicated that OBC may be more toxic than FBC. However, our results suggested that FBC and OBC might lead to diverse toxic endpoints through activating different molecular pathways.


A Comparison of Seven Cox Regression-Based Models to Account for Heterogeneity Across Multiple HIV Treatment Cohorts in Latin America and the Caribbean.
Many studies of HIV/AIDS aggregate data from multiple cohorts to improve power and generalizability. There are several analysis approaches to account for cross-cohort heterogeneity; we assessed how different approaches can impact results from an HIV/AIDS study investigating predictors of mortality. Using data from 13,658 HIV-infected patients starting antiretroviral therapy from seven Latin American and Caribbean cohorts, we illustrate the assumptions of seven readily implementable approaches to account for across cohort heterogeneity with Cox proportional hazards models, and we compare hazard ratio estimates across approaches. As a sensitivity analysis, we modify cohort membership to generate specific heterogeneity conditions. Hazard ratio estimates varied slightly between the seven analysis approaches, but differences were not clinically meaningful. Adjusted hazard ratio estimates for the association between AIDS at treatment initiation and death varied from 2.00 to 2.20 across approaches that accounted for heterogeneity; the adjusted hazard ratio was estimated as 1.73 in analyses that ignored across cohort heterogeneity. In sensitivity analyses with more extreme heterogeneity, we noted a slightly greater distinction between approaches. Despite substantial heterogeneity between cohorts, the impact of the specific approach to account for heterogeneity was minimal in our case study. Our results suggest that it is important to account for across cohort heterogeneity in analyses, but that the specific technique for addressing heterogeneity may be less important. Because of their flexibility in accounting for cohort heterogeneity, we prefer stratification or meta-analysis methods, but we encourage investigators to consider their specific study conditions and objectives.


Host resistance and pathogen infectivity in host populations with varying connectivity.
Theory predicts that hosts and pathogens will evolve higher resistance and aggressiveness in systems where populations are spatially connected than in situations in which populations are isolated and dispersal is more local. In a large cross-inoculation experiment we surveyed patterns of host resistance and pathogen infectivity in anther-smut diseased Viscaria alpina populations from three contrasting areas where populations range from continuous, through patchy but spatially connected to highly isolated demes. In agreement with theory, isolated populations of V. alpina were more susceptible on average than either patchily distributed or continuous populations. While increased dispersal in connected systems increases disease spread, it may also increase host gene flow and the potential for greater host resistance to evolve. In the Viscaria-Microbotryum system, pathogen infectivity mirrored patterns of host resistance with strains from the isolated populations being the least infective and strains from the more resistant continuous populations being the most infective on average, suggesting that high resistance selects for high infectivity. To our knowledge this study is the first to characterize the impacts of varying spatial connectivity on patterns of host resistance and pathogen infectivity in a natural system.


Lower hospital volume is associated with higher mortality after oesophageal, gastric, pancreatic and rectal cancer resection.
In various countries, the association of lower hospital volume and higher mortality after oesophageal, gastric, pancreatic and rectal cancer resection has been clearly demonstrated. However, scientific evidence regarding the volume-outcomes relationship for high-risk visceral surgical procedures in Switzerland is lacking. The a priori hypothesis of this retrospective population-based cohort study analysis was that low-volume hospitals in Switzerland have a higher rate of postoperative mortality after oesophageal, gastric, pancreatic and rectal cancer resection. Patients undergoing elective resection of oesophageal, gastric, pancreatic and rectal cancer between 1999 and 2012 were identified in the inpatient database of the Swiss Federal Statistical Office. Nonparametric correlation analyses were used to assess time trends. Mortality was assessed in univariable and risk-adjusted conditional logistic regression analyses with stratification for year of surgery. A total of 1487 oesophageal, 4404 gastric, 2668 pancreatic and 9743 rectal cancer patients were identified. For all cancer entities, significant treatment centralisation was observed over the time period (all p <0.001). The rate of mortality was inversely related to the annual number of patients treated at a certain hospital. The decrease of postoperative mortality from low-volume to high-volume hospitals was 6.3% to 3.3% for oesophageal cancer (p = 0.019), 4.9% to 3.3% for gastric cancer (p = 0.023), 5.4% to 2.0% for pancreatic cancer (p = 0.037), and 2.4% to 1.6% for rectal cancer (p = 0.008). These results were confirmed in risk-adjusted analyses with a decreased odds of pos-operative death by 49% for oesophageal (odds ratio [OR] 0.51, 95% confidence interval [CI] 0.22-1.18; p = 0.085), 32% for gastric (OR 0.68, 95% CI 0.48-0.98; p = 0.032), 68% for pancreatic (OR 0.32, 95% CI 0.11-0.89; p = 0.011) and 29% for rectal cancer (OR 0.71, 95% CI 0.52-0.98; p = 0.033). This population-based analysis - the first of its kind in the literature - demonstrates a higher postoperative mortality in low-volume hospitals for patients undergoing oesophageal, gastric, pancreatic and rectal cancer resection in Switzerland. Hence, such operations should preferably be performed in high-volume hospitals.


Euglycemic hyperinsulinemia augments the cytokine and endocrine responses to endotoxin in humans.
Type 2 diabetes is associated with biochemical evidence of low-grade inflammation, and experimental studies have suggested that both insulin and glucose affect inflammatory responses. To determine the effect of in vivo changes in glucose availability and plasma insulin concentrations in humans, we administered 20 U/kg Escherichia coli lipopolysaccharide (LPS) or saline (control) to 14 subjects during a euglycemic hyperinsulinemic clamp (n = 6) or an infusion of sterile saline (n = 8). Parallel in vitro studies on human whole blood were undertaken to determine whether there was a direct effect of glucose, insulin, and leptin on proinflammatory cytokine production. Infusion of glucose and insulin significantly amplified and/or prolonged the cardiovascular, plasma interleukin-6 (IL-6), tumor necrosis factor-alpha (TNF-alpha), and counterregulatory hormone responses to LPS, whereas the effects on fever, plasma norepinephrine concentrations, and oxygen consumption were unaffected. In vitro studies showed no modulation of LPS-stimulated IL-6 or TNF-alpha production by glucose, insulin, or leptin at physiologically relevant concentrations. Hyperinsulinemia indirectly enhances key components of the systemic inflammatory and stress responses in this human model of infection.


Three-dimensional scheme for time-domain fluorescence molecular tomography based on Laplace transforms with noise-robust factors.
As a visualizing and quantitative method, Fluorescence Molecular Tomography (FMT) has many potential applications in biomedical field and its three-dimensional (3D) implementation is needed in both theory and practice. In this paper, we propose a 3D scheme for time-domain FMT within the normalized Born-ratio formulation. A finite element method solution to the Laplace transformed time-domain coupled diffusion equations is employed as the forward model, and the resultant linear inversions at two distinct transform-factors are solved with an algebraic reconstruction technique to separate fluorescent yield and lifetime images. The algorithm is validated using simulated data for 3D cylinder phantoms, and the spatial resolution and quantitativeness of the reconstruction assessed. We demonstrate that the proposed approach can accurately retrieve the positions and shapes of the targets with high spatial resolution and quantitative accuracy, and tolerate a signal-to-noise ratio down to 25dB by appropriately choosing the transform factors.


TrichOME: a comparative omics database for plant trichomes.
Plant secretory trichomes have a unique capacity for chemical synthesis and secretion and have been described as biofactories for the production of natural products. However, until recently, most trichome-specific metabolic pathways and genes involved in various trichome developmental stages have remained unknown. Furthermore, only a very limited amount of plant trichome genomics information is available in scattered databases. We present an integrated "omics" database, TrichOME, to facilitate the study of plant trichomes. The database hosts a large volume of functional omics data, including expressed sequence tag/unigene sequences, microarray hybridizations from both trichome and control tissues, mass spectrometry-based trichome metabolite profiles, and trichome-related genes curated from published literature. The expressed sequence tag/unigene sequences have been annotated based upon sequence similarity with popular databases (e.g. Gene Ontology, Kyoto Encyclopedia of Genes and Genomes, and Transporter Classification Database). The unigenes, metabolites, curated genes, and probe sets have been mapped against each other to enable comparative analysis. The database also integrates bioinformatics tools with a focus on the mining of trichome-specific genes in unigenes and microarray-based gene expression profiles. TrichOME is a valuable and unique resource for plant trichome research, since the genes and metabolites expressed in trichomes are often underrepresented in regular non-tissue-targeted cDNA libraries. TrichOME is freely available at http://www.planttrichome.org/.


Bereitschaftspotential in depressed and non-depressed patients with Parkinson's disease.
Impaired initiation and slowed execution of movements are two of the principal characteristics of Parkinson's disease (PD). A similar pattern of movement impairments (psychomotor retardation) can be seen frequently in patients with idiopathic depression. In addition, affective disorders have been frequently reported in patients with different basal ganglia disorders. The aim of this study was to determine whether there are some particularities in the cerebral electrical activity during the preparation and execution of voluntary internally paced movements (i.e., Bereitschaftspotential, BP) in depressed PD patients, which can distinguish them from non-depressed PD patients, as well as from healthy controls. The BPs were recorded in 16 patients with idiopathic PD, eight of whom were depressed (PD-D), and eight of whom were not (PD-ND). Additional recordings were taken from a group of eight age- and sex-matched healthy subjects. Depression was classified using the Research Diagnostic Criteria and the two PD groups were matched for age, disease severity, and disease duration. The amplitudes and slopes of the BPs from PD patients were generally smaller than in controls, but there was no specific pattern of BP changes that distinguished depressed from non-depressed PD patients. In addition, there was no particular association between measures of depression severity and BP parameters. The data suggest that presence of depression in PD might not have any additional deteriorating influence on already impaired preparation for self-paced spontaneous movements.


Endothelial NO synthase phosphorylated at SER635 produces NO without requiring intracellular calcium increase.
Shear stress stimulates NO production involving the Ca2+-independent mechanisms in endothelial cells. We have shown that exposure of bovine aortic endothelial cells (BAEC) to shear stress stimulates phosphorylation of eNOS at S635 and S1179 by the protein kinase A- (PKA-) dependent mechanisms. We examined whether phosphorylation of S635 of eNOS induced by PKA stimulates NO production in a calcium-independent manner. Expression of a constitutively active catalytic subunit of PKA (Cqr) in BAEC induced phosphorylation of S635 and S1179 residues and dephosphorylation of T497. Additionally, Cqr expression stimulated NO production, which could not be prevented by treating cells with the intracellular calcium chelator BAPTA-AM. To determine the role of each eNOS phosphorylation site in NO production, HEK-293 cells transfected with eNOS point mutants whereby S116, T497, S635, and S1179 were mutated to either A or D. Maximum NO production from S635D-expressing cells was significantly higher than that of either wild type or S635A in both basal and elevated [Ca2+]i conditions. More interestingly, S635D cells produced NO even when [Ca2+]i was nearly depleted by BAPTA-AM. We confirmed these results obtained in HEK-293 cells in BAEC transfected with S635D, S635A, or wild-type eNOS vector. These findings suggest that, once phosphorylated at S635 residue, eNOS produces NO without requiring any changes in [Ca2+]i. PKA-dependent phosphorylation of eNOS S635 and subsequent basal NO production in a Ca2+-independent manner may play an important role in regulating vascular biology and pathophysiology.


Stepped Care as an Implementation and Service Delivery Model for Cognitive Behavioral Therapy for Psychosis.
Cognitive behavioral therapy for schizophrenia spectrum disorders is an evidence-based treatment that is recommended by United States schizophrenia treatment guidelines. Based on recent estimates, only 0.3% of individuals with a primary psychotic disorder are able to access this treatment in the United States. Stepped care interventions have shown promise as an applied treatment delivery model in other settings and for other psychotherapeutic interventions. The current paper describes how the stepped care model can be applied to CBT for psychosis in the US to increase access to the intervention in community mental health settings by leveraging the multidisciplinary team.


Voltage-gated sodium channels in cardiac microvascular endothelial cells.
The goal of this study was to determine whether inward Na+ or Ca2+ currents could be measured in cardiac microvascular endothelial cells (CMEC). CMEC were isolated from rat ventricular muscle and studied during days 1-4 in culture. Differential uptake of fluorescently labeled acetylated low-density lipoproteins (LDL) indicated that the primary culture contained > 90% CMEC. Membrane currents were measured with the use of the whole cell arrangement of the patch-clamp technique with a Cs+ internal solution to prevent contamination by outward K+ currents. Voltage steps positive to -30 mV resulted in the activation of a fast, inward Na+ current (INa). In 20 cells examined, the peak inward current measured at 0 mV was 2.1 pA/pF. The half-maximal voltage required for inactivation of INa was -45 mV, and the current recovered from inactivation with a time constant of 10 ms. Inward currents were eliminated by replacement of external sodium with N-methylglucamine and were blocked by both tetrodotoxin (TTX) (dissociation constant = 5 nM) and saxitoxin (50 nM). Stimulation of protein kinase C, through application of phorbol 12,13-dibutyrate, resulted in an increase in the amplitude of INa without any change in the voltage dependence of current activation. Thus the endothelium of cardiac microvessels may be unique in expressing voltage gated, TTX-sensitive Na+ channels.


Effectiveness of rapid diagnostic tests to assess pathogens of fishers (Martes pennanti) and gray foxes (Urocyon cinereoargenteus).
Wildlife managers often need to assess the current health status of wildlife communities before implementation of management actions involving surveillance, reintroductions, or translocations. We estimated the sensitivity and specificity of a commercially available domestic canine rapid diagnostic antigen test for canine parvovirus and a rapid enzyme-linked immunosorbent assay for the detection of antibodies toward Anaplasma phagocytophilum on populations of fishers (Martes pennanti) and sympatric gray foxes (Urocyon cinereoargenteus). Eighty-two fecal samples from 66 fishers and 16 gray foxes were tested with both SNAP((R)) PARVO rapid diagnostic test (RDT) and a nested polymerase chain reaction (PCR). Whole blood samples from 23 fishers and 53 gray foxes were tested with both SNAP 4Dx RDT and immunofluorescence assays (IFAs). The SNAP PARVO RDT detected no parvovirus, whereas PCR detected the virus in 17 samples. Eleven samples were positive using the SNAP 4Dx RDT, whereas 46 samples tested by IFA were positive for A. phagocytophilum. Both RDTs had low sensitivity and poor test agreement. These findings clearly demonstrate the importance of validating RDTs developed for domesticated animals before using them for wildlife populations.


Hunter-Gatherer Social Networks and Reproductive Success.
Individuals' centrality in their social network (who they and their social ties are connected to) has been associated with fertility, longevity, disease and information transmission in a range of taxa. Here, we present the first exploration in humans of the relationship between reproductive success and different measures of network centrality of 39 Agta and 38 BaYaka mothers. We collected three-meter contact ('proximity') networks and reproductive histories to test the prediction that individual centrality is positively associated with reproductive fitness (number of living offspring). Rather than direct social ties influencing reproductive success, mothers with greater indirect centrality (i.e. centrality determined by second and third degree ties) produced significantly more living offspring. However, indirect centrality is also correlated with sickness in the Agta, suggesting a trade-off. In complex social species, the optimisation of individuals' network position has important ramifications for fitness, potentially due to easy access to different parts of the network, facilitating cooperation and social influence in unpredictable ecologies.


Pitfalls in bone mineral density measurements.
Bone mineral density (BMD) measurements have been the single greatest advancement for osteoporosis. BMD measurements have helped define a prefracture diagnosis of osteoporosis, predict fracture risk in postmenopausal women and elderly men, and monitor the course of disease processes that negatively affect bone or therapeutic agents that can improve bone strength. Despite the large amount of clinical, epidemiologic, and basic science data that has advanced our understanding of BMD performance and interpretation, many pitfalls in BMD performance and interpretation pervade the practice of bone densitometry. However, all of these pitfalls can be overcome. Proper quality control and clinical interpretation of BMD computer printout reports are paramount for correct diagnosis, risk assessment, and serial BMD measurements. Though BMD application(s) are a clinical tool that can and should be used by many different primary care and specialty physicians, the performances and interpretations are not simple processes. Proper education and training in the use of BMD technologies provides the means of achieving the great intent that BMD measurements are capable of providing.


Melatonin for Acute Treatment of Migraine in Children and Adolescents: A Pilot Randomized Trial.
To determine what dose of melatonin is most effective for treating migraine acutely in children and adolescents. Acute migraine medications may not work for all patients and may cause side effects. Melatonin is effective for migraine prevention in adults and has been used acutely for procedural pain in children. Our goal was to determine whether a "high" or "low" dose of melatonin is more effective for treating migraine acutely in youth. In this pilot, randomized, open-label, single-center, dose-finding trial, children and adolescents aged 4-17 years with episodic migraine were randomized to "high-dose" or "low-dose" dose melatonin (<40 kg: 4 mg vs. 1 mg; ≥40 kg: 8 mg vs. 2 mg). The primary outcome measure was change in mean pain score between time 0 and 2 hours. Secondary outcomes included 2-hour pain-relief and pain-freedom rates. Eighty-four participants (n = 42 per group) were enrolled in this study. Mean (SD) participant age was 11.8 (3.5) years and 55% (46/84) were female. Mean (SD) headache days/month was 5.6 (3.8). Sixty-six (79%) participants provided outcome data and were included in the analyses, n = 24 in the high-dose group and n = 22 in the low-dose group. The drop-out rate was 43% (18/42) in the high-dose group vs. 48% (20/42) in the low-dose group. Mean (SD) change in pain intensity at 2 hours was -2.7 (2.1) cm in the high-dose group vs. -2.3 (2.1) cm in the low-dose group (p = .581), a difference of 0.4 cm (95% CI: -1.17 to 1.92). Two-hour pain-freedom rate was 41% (7/17) vs. 27% (4/15) in the high-dose vs. low-dose groups (p = .415), and 2-hour pain-relief rate was 94% (16/17) vs. 80% (12/15), (p = .482). There were no serious adverse events. Napping occurred in the majority (67% (14/21) high dose vs. 47% (9/19) low dose). Higher mg/kg dose of melatonin and napping were each independently associated with greater headache benefit. As an acute treatment for pediatric migraine, both low and high doses of melatonin were associated with pain reduction; however, study drop-out was high. Higher dose and napping after treatment predicted greater benefit.


Mutations in the G-quadruplex silencer element and their relationship to c-MYC overexpression, NM23 repression, and therapeutic rescue.
We have demonstrated that a parallel G-quadruplex structure in the c-MYC promoter functions as a transcriptional repressor element. Furthermore, a specific G-to-A mutation in this element results in destabilization of the G-quadruplex repressor element and an increase in basal transcriptional activity. To validate this model in an in vivo context, we have examined the sequence of this region in human colorectal tumors and the surrounding normal tissue. We have found that approximately 30% of tumors contain one of two specific G-to-A mutations, not present in the surrounding normal tissue, that destabilize the parallel G-quadruplex, which would be expected to give rise to abnormally high expression of c-MYC in these cells. In contrast, G-quadruplex-disruptive mutations were absent in 20 colon adenomas, suggesting that these mutations occur late in tumorigenesis. We have also demonstrated that these same mutations are found in established colorectal cell lines. NM23-H2 levels are lower in cancer tissues and cell lines that harbor these mutations. In cells with repressed levels of NM23-H2, the mutated and destabilized G-quadruplex silencer element can be reinstated by the addition of G-quadruplex-stabilizing compounds, providing an opportunity for therapeutic intervention for patients carrying these mutations.


Maternal and obstetrical outcome in 35 cases of well-differentiated thyroid carcinoma during pregnancy.
Thyroid cancer, with 6% to 10% of cancer diagnoses, is one of the most common malignancies during pregnancy. Its treatment poses a risk for the pregnancy, as the thyroid gland plays a crucial role in the evolution of pregnancy. The aim of this study is to evaluate treatment of primary well-differentiated thyroid carcinoma during pregnancy and fetal and maternal outcomes. This is an international cohort study. Primary thyroid cancer patients were identified from the database of the International Network on Cancer, Infertility, and Pregnancy registration study. Data on histopathological characteristics, diagnostic and therapeutic interventions, outcome (obstetrical, neonatal, and maternal) and maternal follow-up were analyzed. Thirty-five patients with well-differentiated thyroid carcinoma were eligible. All 35 patients underwent surgery, 29 (83%) of which during pregnancy. Procedures during pregnancy were mainly total thyroidectomies (n = 24). The median number of days between diagnosis and surgical treatment was different between the groups with surgery during and after pregnancy (27 vs. 139 days, P < .001). Both maternal and neonatal outcomes were uncomplicated, regardless of gestational age during surgery. Well-differentiated thyroid carcinoma diagnosed during pregnancy has a favorable outcome for both mother and child. Surgical management during pregnancy has no negative impact on the pregnancy regardless of the trimester at the time of surgery. However, the potential negative effects of thyroid surgery early in pregnancy demand management of these patients in an experienced multidisciplinary team to provide the best possible care for these patients and their unborn babies. 4. Laryngoscope, 128:1493-1500, 2018.