Systematic and quantitative mRNA expression analysis of TRP channel genes at the single trigeminal and dorsal root ganglion level in mouse.
Somatosensory nerve fibres arising from cell bodies within the trigeminal ganglia (TG) in the head and from a string of dorsal root ganglia (DRG) located lateral to the spinal cord convey endogenous and environmental stimuli to the central nervous system. Although several members of the transient receptor potential (TRP) superfamily of cation channels have been implicated in somatosensation, the expression levels of TRP channel genes in the individual sensory ganglia have never been systematically studied. Here, we used quantitative real-time PCR to analyse and compare mRNA expression of all TRP channels in TG and individual DRGs from 27 anatomically defined segments of the spinal cord of the mouse. At the mRNA level, 17 of the 28 TRP channel genes, TRPA1, TRPC1, TRPC3, TRPC4, TRPC5, TRPM2, TRPM3, TRPM4, TRPM5, TRPM6, TRPM7, TRPM8, TRPV1, TRPV2, TRPV4, TRPML1 and TRPP2, were detectable in every tested ganglion. Notably, four TRP channels, TRPC4, TRPM4, TRPM8 and TRPV1, showed statistically significant variation in mRNA levels between DRGs from different segments, suggesting ganglion-specific regulation of TRP channel gene expression. These ganglion-to-ganglion differences in TRP channel transcript levels may contribute to the variability in sensory responses in functional studies. We developed, compared and refined techniques to quantitatively analyse the relative mRNA expression of all TRP channel genes at the single ganglion level. This study also provides for the first time a comparative mRNA distribution profile in TG and DRG along the entire vertebral column for the mammalian TRP channel family.


Vaccinations administered during off-clinic hours at a national community pharmacy: implications for increasing patient access and convenience.
Approximately 50,000 adults die annually from vaccine-preventable diseases in the United States. Most traditional vaccine providers (eg, physician offices) administer vaccinations during standard clinic hours, but community pharmacies offer expanded hours that allow patients to be vaccinated at convenient times. We analyzed the types of vaccines administered and patient populations vaccinated during off-clinic hours in a national community pharmacy, and their implications for vaccination access and convenience. We retrospectively reviewed data for all vaccinations given at the Walgreens pharmacy chain between August 2011 and July 2012. The time of vaccination was categorized as occurring during traditional hours (9:00 am-6:00 pm weekdays) or off-clinic hours, consisting of weekday evenings, weekends, and federal holidays. We compared demographic characteristics and types of vaccine. We used a logistic regression model to identify predictors of being vaccinated during off-clinic hours. During the study period, pharmacists administered 6,250,402 vaccinations, of which 30.5% were provided during off-clinic hours: 17.4% were provided on weekends, 10.2% on evenings, and 2.9% on holidays. Patients had significantly higher odds of off-clinic vaccination if they were younger than 65 years of age, were male, resided in an urban area, and did not have any chronic conditions. A large proportion of adults being vaccinated receive their vaccines during evening, weekend, and holiday hours at the pharmacy, when traditional vaccine providers are likely unavailable. Younger, working-aged, healthy adults, in particular, a variety of immunizations during off-clinic hours. With the low rates of adult and adolescent vaccination in the United States, community pharmacies are creating new opportunities for vaccination that expand access and convenience.


Bioremediation of oil by marine microbial mats.
Cyanobacterial mats developing in oil-contaminated sabkhas along the African coasts of the Gulf of Suez and in the pristine Solar Lake, Sinai, were collected for laboratory studies. Samples of both mats showed efficient degradation of crude oil in the light, followed by development of an intense bloom of Phormidium spp. and Oscillatoria spp. Isolated cyanobacterial strains, however, did not degrade crude oil in axenic cultures. Strains of sulfate-reducing bacteria and aerobic heterotrophs were capable of degrading model compounds of aliphatic and aromatic hydrocarbons. Results indicate that degradation of oil was done primarily by aerobic heterotrophic bacteria. The oxygenic photosynthesis of oil-insensitive cyanobacteria supplied the molecular oxygen for the efficient aerobic metabolism of organisms, such as Marinobacter sp. The diurnal shifts in environmental conditions at the mat surface, from highly oxic conditions in the light to anaerobic sulfide-rich habitat in the dark, may allow the combined aerobic and anaerobic degradation of crude oil at the mat surface. Hence, coastal cyanobacterial mats may be used for the degradation of coastline oil spills. Oxygen microelectrodes detected a significant inhibition of photosynthetic activity subsequent to oil addition. This prevailed for a few hours and then rapidly recovered. In addition, shifts in bacterial community structure following exposure to oil were determined by denaturing gradient gel electrophoresis of PCR-amplified fractions of 16S rRNA from eubacteria, cyanobacteria and sulfate-reducing bacteria. Since the mats used for the present study were obtained from oil-contaminated environments, they were believed to be preequilibrated for petroleum remediation. The mesocosm system at Eilat provided a unique opportunity to study petroleum degradation by mats formed under different salinities (up to 21%). These mats, dominated by cyanobacteria, can serve as close analogues to the sabkhas contaminated during the Gulf War in Kuwait and Saudi Arabia.


Association between length of residence and overweight among adult immigrants in Portugal: A nationwide cross-sectional study.
Despite the importance of immigrant population in Portugal few studies have analyzed the patterns of overweight/obesity in this subpopulation. The aims of this study are: (i) describe and compare the prevalence of overweight between immigrants and natives in Portugal; (ii) analyze the association between length of residence and overweight among adult immigrants in Portugal. A cross-sectional study (2005-2006) in a representative sample of the Portuguese population from national territory, including the Autonomous Regions of Azores and Madeira. The final sample comprised 31,685 adult participants (≥19 years old), of whom 4.6% were immigrants. Country of birth was used to determine immigrant condition. Logistic regressions were conducted to investigate the association between overweight (dependent variable) and length of residence (exposure), adjusting for all covariates in the study. A 5% confidence level and 95% CI were considered. The percentage of immigrants that are overweight [44.9% (95% CI: 42.3; 47.5)] was lower than for natives [52.8% (95% CI: 52.2; 53.4)]. The migrant condition, after adjusted for sociodemographic variables, was not associated with overweight [OR 1.004 (95% CI: 0.998; 1.010)]. Among immigrants, being women [OR 0.585 (95% CI: 0.583; 0.587)], not married [OR 0.784 (95% CI: 0.781; 0.787)] and with a higher education [OR 0.481 (95% CI: 0.478; 0.483)], are probably protective factors of being overweight. Adjusting for other factors, the odds of being overweight for a long-term immigrant (≥15 years) was 1.3 times higher [OR 1.274 (95% CI: 1.250; 1.299)] than for the newcomers (<4 years). The prevalence of overweight was higher for natives than immigrants. Length of residence (≥15 years) was positively associated with prevalence of overweight, among adult immigrant population. In the future, understanding dietary patterns and acculturation process may be important for health immigrant studies.


Antimutagenic activity of extracts from anticancer drugs in Chinese medicine.
The antimutagenic activities of extracts of 36 commonly used anticancer crude drugs from Chinese herbs were studied by using the Salmonella/microsomal system in the presence of picrolonic acid or benzo[a]pyrene to test whether they contain direct or indirect antimutagens. Each crude drug was extracted with boiling water for 2 h, the method which is commonly used by Chinese people to prepare the drug for oral intake. The extracts of Pteris multifida P. showed the highest antimutagenic activity against picrolonic acid-induced mutation. The extracts of 6 other different kinds of Chinese herbs were shown to have a moderate antimutagenic activity against picrolonic acid-induced mutation, and they are: Actinidia chinensis P., Artemisia lavendulaefolia DC. and Crotalaria sessiflora L., Prunella vulgaris L., Paris polyphylla S. and Ampelopsis brevipedunculata T. The extracts of Smilax china L., Prunella vulgaris L. and Actinidia chinensis P. were demonstrated to inhibit the mutagenicity of benzo[a]pyrene completely. The 12 other kinds of extracts of Chinese herbs which had a moderate antimutagenic activity against benzo[a]pyrene were: Pteris polyphylla S., Ampelopsis brevipedunculata T., Duchesnea indica F., Gossypium herbaceum L., Lithospermum erythrorrhizon SZ., Artemisia lavendulaefolia DC., Selaginella doederleinii H., Dianthus superbus L., Centipeda minima ABA., Curcuma zedoaria R., Marsdenia tenacissima WA. and Kalopanax septemlobus K. Among them, there were 5 kinds of crude drugs, Actinidia chinensis P., Artemisia lavendulaefolia DC., Prunella vulgaris L., Paris polyphylla S. and Ampelopsis brevipedunculata T., containing antimutagenic factors against both picrolonic acid- and benzo[a]pyrene-induced mutation.


Advanced glycation end-products suppress neuropilin-1 expression in podocytes.
Advanced glycation end products (AGEs) have been linked to the pathogenesis of diabetic nephropathy. Here we tested the effect of AGE-modified bovine serum albumin (AGE-BSA) on differentiated mouse podocytes in culture. Differential display and real-time PCR analyses showed that in addition to neuropilin-1, the entire signaling receptor complex of neuropilin-2, semaphorin-3A, and plexin-A1, was significantly reduced by AGE-BSA as was neuropilin-1 protein. The effect was specific for podocytes compared to isolated mesangial and tubular epithelial cells. Further, AGE-BSA was not toxic to podocytes. Neuropilin-1 expression was decreased in glomeruli of diabetic db/db mice compared to their non-diabetic littermates. Transcripts of both neuropilins were found to be decreased in renal biopsies from patients with diabetic nephropathy compared to transplant donors. Podocyte migration was inhibited by AGE-BSA with similar results found in the absence of AGE-BSA when neuropilin-1 expression was down-regulated by siRNA. In contrast, podocyte migration was stimulated by overexpression of neuropilin-1 even in the presence of AGE-BSA. Our study shows that AGE-BSA inhibited podocyte migration by down-regulating neuropilin-1. The decreased migration could lead to adherence of uncovered areas of the glomerular basement membrane to Bowman's capsule contributing to focal glomerulosclerosis.


Protein adsorption studies on 'standard' polymeric materials.
Adsorption of fibrinogen from buffer as a single protein and from plasma to four materials has been studied. The two NIH-NHLBI primary reference standards, filler free polydimethylsiloxane and low density polyethylene, were used along with polyvinylcholoride and cellulose materials supplied by the IUPAC Working Party. The materials were examined in both film and tubing form, except for polydimethylsiloxane which was studied only in tubing form. Adsorption was measured at room temperature using 125I-labelled fibrinogen. The order of adsorbed amounts in the single protein experiments was found to be: cellulose < PVC < PE = PDMS. Apparent adsorption affinities are in the same order. In plasma, all surfaces except cellulose showed maxima in adsorption as a function of plasma concentration after 5 min contact. This is indicative of initial adsorption followed by displacement of fibrinogen (the Vroman effect). Cellulose showed very low adsorption of fibrinogen from plasma. The Vroman maxima were more pronounced on the tubing samples than on the films, and, as for the single protein experiments, adsorption was found to be less on tubing than on film samples. A tentative interpretation of the Vroman effect data suggests that the order of procoagulant activity of the materials may be: PDMS = PE < PVC < cellulose.


NACP: Partnership for Native American Cancer Prevention.
Cancer trends over a two-decade period show a greater reduction in cancer mortality rates for non-Hispanic Whites than for Native Americans. The Partnership for Native American Cancer Prevention (NACP) was established to address cancer health disparities that impact Native Americans. The partners are Northern Arizona University, the University of Arizona Cancer Center, Arizona's tribal communities and the National Cancer Institute. The activities include outreach, research and cancer education. Overall, NACP seeks to expand capacity for culturally-sensitive and community-relevant research on cancer, and to continue developing respectful collaborations that will empower sovereign Native American communities to define, implement, and achieve their goals for cancer health equity.


Acupuncture treatment for idiopathic trigeminal neuralgia: A longitudinal case-control double blinded study.
To evaluate the treatment effect of acupuncture on patients with idiopathic trigeminal neuralgia (ITN) by case-control longitudinal blinded study. Sixty ITN patients and 30 healthy subjects were included. The ITN patients were randomly assigned to acupuncture group (15 cases), sham-acupuncture group (15 cases) and carbamazepine group (30 cases), respectively. Clinical orofacial evaluation (including pain intensity and medication doses), research diagnostic criteria for temporomandibular disorders (RDC/TMD) and Helkimo indexes (for functional evaluation of the masticatory system), and quantitative sensory testing for sensory thresholds (gustative, olfactory, cold, warm, touch, vibration and superficial and deep pain) were evaluated before treatment, immediately after treatment, and 6 months after treatment. The mean pain intensity by the Visual Analogue Scale only decreased in the acupuncture group at the last evaluation (P=0.012). Patients in the sham-acupuncture group had an increase in carbamazepine doses according to the prescriptions (P<0.01). There was a reduction in secondary myofascial pain and mandibular limitations at the acupuncture and sham-acupuncture groups, however only the acupuncture group kept the changes after 6 months (P<0.01, P=0.023). There was a decrease in mechanical thresholds in the acupuncture group (tactile, P<0.01; vibration, P=0.027) and an increase in deep pain thresholds in both acupuncture and sham-acupuncture groups (P=0.013). Acupuncture can be an option in the treatment of ITN due to its analgesic effect in both ITN and secondary myofascial pain associated with it.


A service evaluation of patient pathways and care experiences of dentally anxious adult patients.
To investigate the current patient pathways used by dentally anxious adults in Sheffield and identify how the patient experience could be improved. Questionnaires gathered stakeholder perceptions of referral pathways and services for dentally anxious adult patients. Completed questionnaires were returned by 113 dentally anxious patients who had engaged with specialised dental services and 111 general dental practitioners (GDPs) (28% and 52% response rates). The recommendations for improving dental care experience of the anxious were: increased guidance and information to GDPs regarding available care pathways; improved availability of psychological services; and more opportunities and choice for patients in the long-term management of dental anxiety. The findings from the service evaluation suggest ways in which dental services could be developed to improve the care experiences of dentally anxious adult patients.


Gefitinib or chemotherapy for non-small-cell lung cancer with mutated EGFR.
Non-small-cell lung cancer with sensitive mutations of the epidermal growth factor receptor (EGFR) is highly responsive to EGFR tyrosine kinase inhibitors such as gefitinib, but little is known about how its efficacy and safety profile compares with that of standard chemotherapy. We randomly assigned 230 patients with metastatic, non-small-cell lung cancer and EGFR mutations who had not previously received chemotherapy to receive gefitinib or carboplatin-paclitaxel. The primary end point was progression-free survival; secondary end points included overall survival, response rate, and toxic effects. In the planned interim analysis of data for the first 200 patients, progression-free survival was significantly longer in the gefitinib group than in the standard-chemotherapy group (hazard ratio for death or disease progression with gefitinib, 0.36; P<0.001), resulting in early termination of the study. The gefitinib group had a significantly longer median progression-free survival (10.8 months, vs. 5.4 months in the chemotherapy group; hazard ratio, 0.30; 95% confidence interval, 0.22 to 0.41; P<0.001), as well as a higher response rate (73.7% vs. 30.7%, P<0.001). The median overall survival was 30.5 months in the gefitinib group and 23.6 months in the chemotherapy group (P=0.31). The most common adverse events in the gefitinib group were rash (71.1%) and elevated aminotransferase levels (55.3%), and in the chemotherapy group, neutropenia (77.0%), anemia (64.6%), appetite loss (56.6%), and sensory neuropathy (54.9%). One patient receiving gefitinib died from interstitial lung disease. First-line gefitinib for patients with advanced non-small-cell lung cancer who were selected on the basis of EGFR mutations improved progression-free survival, with acceptable toxicity, as compared with standard chemotherapy. (UMIN-CTR number, C000000376.)


Mycolactone suppresses T cell responsiveness by altering both early signaling and posttranslational events.
Mycolactone is a diffusible lipid toxin produced by Mycobacterium ulcerans, the causative agent of a necrotizing skin disease referred to as Buruli ulcer. Intriguingly, patients with progressive lesions display a systemic suppression of Th1 responses that resolves on surgical excision of infected tissues. In this study, we examined the effects of mycolactone on the functional biology of T cells and identified two mechanisms by which mycolactone suppresses cell responsiveness to antigenic stimulation. At noncytotoxic concentrations, mycolactone blocked the activation-induced production of cytokines by a posttranscriptional, mammalian target of rapamycin, and cellular stress-independent mechanism. In addition, mycolactone triggered the lipid-raft association and activation of the Src-family kinase, Lck. Mycolactone-mediated hyperactivation of Lck resulted in the depletion of intracellular calcium stores and downregulation of the TCR, leading to impaired T cell responsiveness to stimulation. These biochemical alterations were not observed when T cells were exposed to other bacterial lipids, or to structurally related immunosuppressors. Mycolactone thus constitutes a novel type of T cell immunosuppressive agent, the potent activity of which may explain the defective cellular responses in Buruli ulcer patients.


The cardiovascular pathology of smoking.
Atherosclerosis, the process underlying coronary heart disease, peripheral vascular disease, and stroke, begins in childhood and progresses through several stages to result in clinically manifest disease in middle age and later. Elevated plasma low-density lipoprotein cholesterol levels, lowered high-density lipoprotein cholesterol levels, and elevated blood pressure are associated with more extensive and more severe atherosclerosis and also with greater risk of clinical disease. Cigarette smoking is firmly established as a risk factor for coronary heart disease and peripheral vascular disease. It is associated with more severe coronary atherosclerosis, but not enough to account for the twofold or greater risk of coronary heart disease among smokers. Smoking is associated with much more severe atherosclerosis of the abdominal aorta, which is consistent with the much greater risk of aortic aneurysm and peripheral vascular disease among smokers. Smoking induces a wide variety of physiologic responses, some of which appear likely to be involved in accelerating atherogenesis or increasing the probability of thrombosis. These responses include reduction in plasma high-density lipoprotein cholesterol concentration, elevation in plasma fibrinogen concentration, and elevation in white blood cell count. The rapid amelioration of the risk of cardiovascular disease after cessation of smoking suggests that these processes are readily reversible.


Removal of atenolol from aqueous solutions by multiwalled carbon nanotubes modified with ozone: kinetic and equilibrium study.
The aim of study is removal of atenolol from aqueous solutions by multiwalled carbon nanotubes modified with ozone. The design of the experiment was adopted across four levels with the L16 matrix arrangement. The factors influencing atenolol adsorption include changes in the pH value, contact time, the dose of the modified multiwall carbon nanotube, and the initial concentration of atenolol in the solution; these factors were evaluated along with the extent of their influence on removal efficiency. Data analyses were performed by the Design Expert 6 software. The results indicated that the pH, contact time, adsorbent dose, and the initial concentration were 7, 20 min, 0.15 g/L and 1 mg/L, respectively. In this state, the removal efficiency was calculated to be 75.79%. The maximum adsorption capacity was obtained as 5.05 mg/g under optimal conditions. The data were analyzed using adsorption models obtained from the isotherm fitting tool software. The results suggested that the data had a greater congruence with the Freundlich model (corrected Akaike information criterion = 2.58). Furthermore, the kinetics of the reactions followed pseudo second order kinetics (R2 = 0.95). Based on this study, it can be concluded that modified multiwall carbon nanotubes enjoy high potential and efficiency as adsorbents for the removal of atenolol from aqueous solutions.


Synaptic patterning and the timescales of cortical dynamics.
Neocortical circuits, as large heterogeneous recurrent networks, can potentially operate and process signals at multiple timescales, but appear to be differentially tuned to operate within certain temporal receptive windows. The modular and hierarchical organization of this selectivity mirrors anatomical and physiological relations throughout the cortex and is likely determined by the regional electrochemical composition. Being consistently patterned and actively regulated, the expression of molecules involved in synaptic transmission constitutes the most significant source of laminar and regional variability. Due to their complex kinetics and adaptability, synapses form a natural primary candidate underlying this regional temporal selectivity. The ability of cortical networks to reflect the temporal structure of the sensory environment can thus be regulated by evolutionary and experience-dependent processes.


A single prophylactic dose of ceftriaxone as total antibiotic therapy in transurethral surgery.
Ceftriaxone (1 g, intravenous administration, 30 to 60 minutes preoperatively) was the only antimicrobial therapy used for 103 male patients undergoing transurethral surgery. Only patients who had sterile urine were included in the study. Patients with a preoperative catheter placed were excluded. One transurethral incision of the prostate, four direct-vision internal urethrotomies, and 98 transurethral resections of the prostate were performed. Urine specimens for culture were obtained preoperatively and on first voiding after catheter removal for all patients. During the first postoperative visit (11 to 40 days after surgery), urinalysis was performed for all patients, and specimens for culture were obtained from 89 patients. No infections were noted on cultures of first voided specimens after catheter removal; there were four infections (3.88%) on first postoperative visit cultures. No significant morbidity occurred secondary to urinary tract infection. According to these findings, the use of antimicrobial agents beyond the preoperative and immediate postoperative period in uninfected patients undergoing transurethral surgery does not appear to be indicated.


A study of temperature rise in the pulp chamber during composite polymerization with different light-curing units.
The study compared pulp temperature rise during polymerization of resin-based composites (RBCs) using halogen and LED light-curing units (LCUs). A total of 32 teeth extracted from patients aged 11-18 years were used in the study. Thermocouples placed on the roof of the pulp chamber using a novel 'split-tooth' method. In Group 1 a halogen LCU with a light intensity of 450 mW cm(-2) was used and in Group 2, an LED LCU with a light intensity of 1100 mW cm(-2) was used. The teeth were placed in a water bath with the temperature regulated until both the pulp temperature and the ambient temperature were stable at 37 degrees C. Continuous temperature records were made via a data logger and computer. The increase in temperature from baseline to maximum was calculated for each specimen during the curing of both the bonding agent and the RBC. The rise in pulp temperature was significantly higher with the LED LCU than with the halogen LCU for bonding and RBC curing (p<0.05). The major rise in temperature occurred during the curing of the bonding agent. During the curing of the RBC, rises were smaller. Curing of bonding agents should be done with low intensity light and high intensity used only for curing RBC regardless of whether LED or halogen LCUs are used.


Ants are the major agents of resource removal from tropical rainforests.
Ants are diverse and abundant, especially in tropical ecosystems. They are often cited as the agents of key ecological processes, but their precise contributions compared with other organisms have rarely been quantified. Through the removal of food resources from the forest floor and subsequent transport to nests, ants play an important role in the redistribution of nutrients in rainforests. This is an essential ecosystem process and a key energetic link between higher trophic levels, decomposers and primary producers. We used the removal of carbohydrate, protein and seed baits as a proxy to quantify the contribution that ants, other invertebrates and vertebrates make to the redistribution of nutrients around the forest floor, and determined to what extent there is functional redundancy across ants, other invertebrate and vertebrate groups. Using a large-scale, field-based manipulation experiment, we suppressed ants from c. 1 ha plots in a lowland tropical rainforest in Sabah, Malaysia. Using a combination of treatment and control plots, and cages to exclude vertebrates, we made food resources available to: (i) the whole foraging community, (ii) only invertebrates and (iii) only non-ant invertebrates. This allowed us to partition bait removal into that taken by vertebrates, non-ant invertebrates and ants. Additionally, we examined how the non-ant invertebrate community responded to ant exclusion. When the whole foraging community had access to food resources, we found that ants were responsible for 52% of total bait removal whilst vertebrates and non-ant invertebrates removed the remaining 48%. Where vertebrates were excluded, ants carried out 61% of invertebrate-mediated bait removal, with all other invertebrates removing the remaining 39%. Vertebrates were responsible for just 24% of bait removal and invertebrates (including ants) collectively removed the remaining 76%. There was no compensation in bait removal rate when ants and vertebrates were excluded, indicating low functional redundancy between these groups. This study is the first to quantify the contribution of ants to the removal of food resources from rainforest floors and thus nutrient redistribution. We demonstrate that ants are functionally unique in this role because no other organisms compensated to maintain bait removal rate in their absence. As such, we strengthen a growing body of evidence establishing ants as ecosystem engineers, and provide new insights into the role of ants in maintaining key ecosystem processes. In this way, we further our basic understanding of the functioning of tropical rainforest ecosystems.


Socio-demographic patterning of referral, uptake and attendance in Physical Activity Referral Schemes.
Inadequate monitoring and participant profiling have so far prevented a detailed examination of who Physical Activity Referral Schemes (PARS) are accessible to and appropriate for. As a result, the nature of the role for PARS within public health is unknown. Participants were all those referred to a countywide PARS during a three-year period (n = 3568). Participant age, gender and the deprivation level and rurality of their area of residence were compared with the average for the county population. Characteristics associated with referral uptake (attending > or =1 exercise session) and completion (> or =80% attendance), were identified using logistic regression. Compared with the county average, participants were older, more predominantly female (61.1 vs 51.4%) and lived in more deprived areas (p < 0.001). Referral uptake (n = 2864) was most likely in those aged 60-69 years, and least likely for residents of rural villages and the most deprived areas (all p < 0.001). For participants who took up referral, completion was most likely in men and the over-seventies (p < 0.001). The PARS format may be inappropriate for younger adults or people living in relative deprivation and rural areas. They appear most appropriate for adults of middle-to-old age who are more likely to require supervision, and should be targeted accordingly.


Primary vs. secondary antibody deficiency: clinical features and infection outcomes of immunoglobulin replacement.
Secondary antibody deficiency can occur as a result of haematological malignancies or certain medications, but not much is known about the clinical and immunological features of this group of patients as a whole. Here we describe a cohort of 167 patients with primary or secondary antibody deficiencies on immunoglobulin (Ig)-replacement treatment. The demographics, causes of immunodeficiency, diagnostic delay, clinical and laboratory features, and infection frequency were analysed retrospectively. Chemotherapy for B cell lymphoma and the use of Rituximab, corticosteroids or immunosuppressive medications were the most common causes of secondary antibody deficiency in this cohort. There was no difference in diagnostic delay or bronchiectasis between primary and secondary antibody deficiency patients, and both groups experienced disorders associated with immune dysregulation. Secondary antibody deficiency patients had similar baseline levels of serum IgG, but higher IgM and IgA, and a higher frequency of switched memory B cells than primary antibody deficiency patients. Serious and non-serious infections before and after Ig-replacement were also compared in both groups. Although secondary antibody deficiency patients had more serious infections before initiation of Ig-replacement, treatment resulted in a significant reduction of serious and non-serious infections in both primary and secondary antibody deficiency patients. Patients with secondary antibody deficiency experience similar delays in diagnosis as primary antibody deficiency patients and can also benefit from immunoglobulin-replacement treatment.


Is optical coherence tomography really a new biomarker candidate in multiple sclerosis?--A structural and functional evaluation.
To assess the structural and functional status of patients with multiple sclerosis (MS) without a history of optic neuritis. Thirty-nine patients with MS who had reported no visual symptoms before and after the time of MS diagnosis were included. Thirty-eight healthy subjects were included as a control group. Retinal nerve fiber layer (RNFL) thickness was determined by optical coherence tomography. Pattern visual evoked potentials (PVEP), full-field electroretinogram (ERG), and multifocal electroretinogram (mfERG) were performed. There was a significant reduction (P = 0.011) only in temporal RNFL thickness in patients with MS. P(100) latency was significantly delayed with both 60-min arc checks (P < 0.001) and 15-min arc checks (P < 0.001); however, P(100) amplitude was significantly reduced only in 60-min arc checks (P = 0.026). Rod response b-wave implicit time and standard combined response a- and b-wave implicit times were significantly delayed in patients with MS. Patients with MS with a delayed P(100) latency (21/39; 53.8%) had significantly reduced cone response b-wave amplitude and significantly delayed cone response a- and b-wave implicit times in ERG. mfERG results did not differ between MS and control subjects and between patients with a delayed and a normal P(100) latency. Pearson correlations between RNFL thickness and P(100) amplitude and latency in patients with MS were not significant (P > 0.05). There is no correlation between RNFL thickness and P(100) response in patients with MS. PVEP seems to be a more reliable biomarker in determining visual pathway involvement in patients with no history of optic neuritis.


Measurement of acetaldehyde in exhaled breath using a laser absorption spectrometer.
A high-resolution liquid-nitrogen-free mid-infrared tunable diode laser absorption spectroscopy (TDLAS) system was used to perform real-time measurement of acetaldehyde concentrations in human exhaled breath following ingestion of an alcoholic beverage. Acetaldehyde absorption features were measured near 5.79 mum (1727 cm(-1)) using a IV-VI semiconductor laser, a 100 m long path optical gas cell, and second- harmonic detection coupled with wavelength modulation. Acetaldehyde levels were measured with a minimum detection limit of 80 ppb for 5 s integration time. The variations in exhaled acetaldehyde levels over time were analyzed prior to and following ingestion of two different amounts of white wine. A method to calibrate acetaldehyde measurements internally using water vapor absorption lines was investigated to eliminate the need for system calibration with gas standards. The potential of a TDLAS system to be used as a noninvasive clinical tool for measurements of large volatile compounds with possible applications in cancer detection is demonstrated.


Multirater agreement of the causes of anterior cruciate ligament reconstruction failure: a radiographic and video analysis of the MARS cohort.
Anterior cruciate ligament (ACL) reconstruction failure occurs in up to 10% of cases. Technical errors are considered the most common cause of graft failure despite the absence of validated studies. Limited data are available regarding the agreement among orthopaedic surgeons regarding the causes of primary ACL reconstruction failure and accuracy of graft tunnel placement. Experienced knee surgeons have a high level of interobserver reliability in the agreement about the causes of primary ACL reconstruction failure, anatomic graft characteristics, and tunnel placement. Cohort study (diagnosis); Level of evidence, 3. Twenty cases of revision ACL reconstruction were randomly selected from the Multicenter ACL Revision Study (MARS) database. Each case included the patient's history, standardized radiographs, and a concise 30-second arthroscopic video taken at the time of revision demonstrating the graft remnant and location of the tunnel apertures. All 20 cases were reviewed by 10 MARS surgeons not involved with the primary surgery. Each surgeon completed a 2-part questionnaire dealing with each surgeon's training and practice, as well as the placement of the femoral and tibial tunnels, condition of the primary graft, and the surgeon's opinion as to the causes of graft failure. Interrater agreement was determined for each question with the kappa coefficient and the prevalence-adjusted, bias-adjusted kappa (PABAK). The 10 reviewers have been in practice an average of 14 years and have performed at least 25 ACL reconstructions per year, and 9 were fellowship trained in sports medicine. There was wide variability in agreement among knee experts as to the specific causes of ACL graft failure. When participants were specifically asked about technical error as the cause for failure, interobserver agreement was only slight (PABAK = 0.26). There was fair overall agreement on ideal femoral tunnel placement (PABAK = 0.55) but only slight agreement on whether a femoral tunnel was too anterior (PABAK = 0.24) and fair agreement on whether it was too vertical (PABAK = 0.46). There was poor overall agreement for ideal tibial tunnel placement (PABAK = 0.17). This study suggests that more objective criteria are needed to accurately determine the causes of primary ACL graft failure as well as the ideal femoral and tibial tunnel placement in patients undergoing revision ACL reconstruction.


Sonographic characteristics of the cavum velum interpositum.
Differential considerations for pineal region CSF collections include both true cysts and normal cystlike anatomic variations. Our purpose was to determine the sonographic characteristics of pineal region fluid spaces that reflect the presence of a normal persistent cavum velum interpositum (CVI). Eighteen neonates and infants who had sonographic findings of "cysts" in the pineal region were examined prospectively with conventional sonography and color Doppler sonography to evaluate the shape of the fluid collection and its anatomic relationship with the color-coded internal cerebral veins. Subsequent MR images were obtained in eight of these infants to determine the exact nature of the cystlike collections. The cystlike spaces in the pineal region were of an inverted helmet shape in 14 subjects and roundish in four. All were situated inferior or slightly anteroinferior to the splenium of the corpus callosum and 2.5 to 4 mm away from the quadrigeminal plate. The internal cerebral veins were either inferior (n = 12) or inferolateral (n = 6) to the cystlike spaces at sonography. Subsequent MR studies confirmed eight of these cystlike spaces to be the posterior portion of the CVI. The CVI may appear as a cyst in the pineal region on neonatal sonograms. Usually, it has a characteristic inverted helmet shape and is situated beneath the fornices and above the internal cerebral veins.


The use of phase-inverting emulsions to show the phenomenon of interfacial crystallization on both heating and cooling.
Highly anomalous crystallization behavior has been achieved in phase-inverting emulsion systems by using nonionic surfactants that induce nucleation. In particular, nucleation can be inhibited at the phase inversion, allowing systems held at, or near, this temperature to undergo crystallization either on heating or cooling. This new phenomenon is demonstrated for 27.4 wt % aqueous glycine solutions emulsified in decane using Span 20 Tween 20 blends. The inhibitory effect on interfacial nucleation at/near the phase inversion is readily shown by the maximum in the induction time for crystallization found in systems at/near the phase-inversion temperature. These findings are unprecedented. An extremely rapid rise in nucleation rate is expected on cooling glycine solutions, owing to the associated increase in supersaturation, the driving force for crystallization. The origin of this highly anomalous behavior is thought to be the low droplet interfacial tension, gammaow, that occurs at the phase-inversion temperature, which results primarily in a substantially increased contact angle between the glycine critical nucleus and the droplet interface. This may present a paradigm shift in crystallization strategies through the use of tunable contact-angle nucleators.


A phase I randomized clinical trial of candidate human immunodeficiency virus type 1 vaccine MVA.HIVA administered to Gambian infants.
A vaccine to decrease transmission of human immunodeficiency virus type 1 (HIV-1) during breast-feeding would complement efforts to eliminate infant HIV-1 infection by antiretroviral therapy. Relative to adults, infants have distinct immune development, potentially high-risk of transmission when exposed to HIV-1 and rapid progression to AIDS when infected. To date, there have been only three published HIV-1 vaccine trials in infants. We conducted a randomized phase I clinical trial PedVacc 001 assessing the feasibility, safety and immunogenicity of a single dose of candidate vaccine MVA.HIVA administered intramuscularly to 20-week-old infants born to HIV-1-negative mothers in The Gambia. Infants were followed to 9 months of age with assessment of safety, immunogenicity and interference with Expanded Program on Immunization (EPI) vaccines. The trial is the first stage of developing more complex prime-boost vaccination strategies against breast milk transmission of HIV-1. From March to October 2010, 48 infants (24 vaccine and 24 no-treatment) were enrolled with 100% retention. The MVA.HIVA vaccine was safe with no difference in adverse events between vaccinees and untreated infants. Two vaccine recipients (9%) and no controls had positive ex vivo interferon-γ ELISPOT assay responses. Antibody levels elicited to the EPI vaccines, which included diphtheria, tetanus, whole-cell pertussis, hepatitis B virus, Haemophilus influenzae type b and oral poliovirus, reached protective levels for the vast majority and were similar between the two arms. A single low-dose of MVA.HIVA administered to 20-week-old infants in The Gambia was found to be safe and without interference with the induction of protective antibody levels by EPI vaccines, but did not alone induce sufficient HIV-1-specific responses. These data support the use of MVA carrying other transgenes as a boosting vector within more complex prime-boost vaccine strategies against transmission of HIV-1 and/or other infections in this age group. ClinicalTrials.gov NCT00982579. The Pan African Clinical Trials Registry PACTR2008120000904116.


Clean home-delivery in rural Southern Tanzania: barriers, influencers, and facilitators.
The study explored the childbirth-related hygiene and newborn care practices in home-deliveries in Southern Tanzania and barriers to and facilitators of behaviour change. Eleven home-birth narratives and six focus group discussions were conducted with recently-delivering women; two focus group discussions were conducted with birth attendants. The use of clean cloth for delivery was reported as common in the birth narratives; however, respondents did not link its use to newborn's health. Handwashing and wearing of gloves by birth attendants varied and were not discussed in terms of being important for newborn's health, with few women giving reasons for this behaviour. The lack of handwashing and wearing of gloves was most commonly linked to the lack of water, gloves, and awareness. A common practice was the insertion of any family member's hands into the vagina of delivering woman to check labour progress before calling the birth attendant. The use of a new razor blade to cut the cord was near-universal; however, the cord was usually tied with a used thread due to the lack of knowledge and the low availability of clean thread. Applying something to the cord was near-universal and was considered essential for newborn's health. Three hygiene practices were identified as needing improvement: family members inserting a hand into the vagina of delivering woman before calling the birth attendant, the use of unclean thread, and putting substances on the cord. Little is known about families conducting internal checks of women in labour, and more research is needed before this behaviour is targeted in interventions. The use of clean thread as cord-tie appears acceptable and can be addressed, using the same channels and methods that were used for successfully encouraging the use of new razor blade.


Bacteriophage T4 32 protein is required for helicase-dependent leading strand synthesis when the helicase is loaded by the T4 59 helicase-loading protein.
In the bacteriophage T4 DNA replication system, T4 gene 59 protein binds preferentially to fork DNA and accelerates the loading of the T4 41 helicase. 59 protein also binds the T4 32 single-stranded DNA-binding protein that coats the lagging strand template. Here we explore the function of the strong affinity between the 32 and 59 proteins at the replication fork. We show that, in contrast to the 59 helicase loader, 32 protein does not bind forked DNA more tightly than linear DNA. 32 protein displays a strong binding polarity on fork DNA, binding with much higher affinity to the 5' single-stranded lagging strand template arm of a model fork, than to the 3' single-stranded leading strand arm. 59 protein promotes the binding of 32 protein on forks too short for cooperative binding by 32 protein. We show that 32 protein is required for helicase-dependent leading strand DNA synthesis when the helicase is loaded by 59 protein. However, 32 protein is not required for leading strand synthesis when helicase is loaded, less efficiently, without 59 protein. Leading strand synthesis by wild type T4 polymerase is strongly inhibited when 59 protein is present without 32 protein. Because 59 protein can load the helicase on forks without 32 protein, our results are best explained by a model in which 59 helicase loader at the fork prevents the coupling of the leading strand polymerase and the helicase, unless the position of 59 protein is shifted by its association with 32 protein.


Web-Based Immersive Virtual Patient Simulators: Positive Effect on Clinical Reasoning in Medical Education.
Clinical reasoning is based on the declarative and procedural knowledge of workflows in clinical medicine. Educational approaches such as problem-based learning or mannequin simulators support learning of procedural knowledge. Immersive patient simulators (IPSs) go one step further as they allow an illusionary immersion into a synthetic world. Students can freely navigate an avatar through a three-dimensional environment, interact with the virtual surroundings, and treat virtual patients. By playful learning with IPS, medical workflows can be repetitively trained and internalized. As there are only a few university-driven IPS with a profound amount of medical knowledge available, we developed a university-based IPS framework. Our simulator is free to use and combines a high degree of immersion with in-depth medical content. By adding disease-specific content modules, the simulator framework can be expanded depending on the curricular demands. However, these new educational tools compete with the traditional teaching It was our aim to develop an educational content module that teaches clinical and therapeutic workflows in surgical oncology. Furthermore, we wanted to examine how the use of this module affects student performance. The new module was based on the declarative and procedural learning targets of the official German medical examination regulations. The module was added to our custom-made IPS named ALICE (Artificial Learning Interface for Clinical Education). ALICE was evaluated on 62 third-year students. Students showed a high degree of motivation when using the simulator as most of them had fun using it. ALICE showed positive impact on clinical reasoning as there was a significant improvement in determining the correct therapy after using the simulator. ALICE positively impacted the rise in declarative knowledge as there was improvement in answering multiple-choice questions before and after simulator use. ALICE has a positive effect on knowledge gain and raises students' motivation. It is a suitable tool for supporting clinical education in the blended learning context.


miRNA expression profiling of 51 human breast cancer cell lines reveals subtype and driver mutation-specific miRNAs.
Breast cancer is a genetically and phenotypically complex disease. To understand the role of miRNAs in this molecular complexity, we performed miRNA expression analysis in a cohort of molecularly well-characterized human breast cancer cell lines to identify miRNAs associated with the most common molecular subtypes and the most frequent genetic aberrations. Using a microarray carrying LNA™ modified oligonucleotide capture probes), expression levels of 725 human miRNAs were measured in 51 breast cancer cell lines. Differential miRNA expression was explored by unsupervised cluster analysis and was then associated with the molecular subtypes and genetic aberrations commonly present in breast cancer. Unsupervised cluster analysis using the most variably expressed miRNAs divided the 51 breast cancer cell lines into a major and a minor cluster predominantly mirroring the luminal and basal intrinsic subdivision of breast cancer cell lines. One hundred and thirteen miRNAs were differentially expressed between these two main clusters. Forty miRNAs were differentially expressed between basal-like and normal-like/claudin-low cell lines. Within the luminal-group, 39 miRNAs were associated with ERBB2 overexpression and 24 with E-cadherin gene mutations, which are frequent in this subtype of breast cancer cell lines. In contrast, 31 miRNAs were associated with E-cadherin promoter hypermethylation, which, contrary to E-cadherin mutation, is exclusively observed in breast cancer cell lines that are not of luminal origin. Thirty miRNAs were associated with p16INK4 status while only a few miRNAs were associated with BRCA1, PIK3CA/PTEN and TP53 mutation status. Twelve miRNAs were associated with DNA copy number variation of the respective locus. Luminal-basal and epithelial-mesenchymal associated miRNAs determine the subdivision of miRNA transcriptome of breast cancer cell lines. Specific sets of miRNAs were associated with ERBB2 overexpression, p16INK4a or E-cadherin mutation or E-cadherin methylation status, which implies that these miRNAs may contribute to the driver role of these genetic aberrations. Additionally, miRNAs, which are located in a genomic region showing recurrent genetic aberrations, may themselves play a driver role in breast carcinogenesis or contribute to a driver gene in their vicinity. In short, our study provides detailed molecular miRNA portraits of breast cancer cell lines, which can be exploited for functional studies of clinically important miRNAs.


Insertion of linear 8.4 μm diameter 16 channel carbon fiber electrode arrays for single unit recordings.
Single carbon fiber electrodes (d = 8.4 μm) insulated with parylene-c and functionalized with pTS have been shown to record single unit activity but manual implantation of these devices with forceps can be difficult. Without an improvement in the insertion method any increase in the channel count by fabricating carbon fiber arrays would be impractical. In this study, we utilize a water soluble coating and structural backbones that allow us to create, implant, and record from fully functionalized arrays of carbon fibers with ∼150 μm pitch. Two approaches were tested for the insertion of carbon fiber arrays. The first method used a poly(ethylene glycol) (PEG) coating that temporarily stiffened the fibers while leaving a small portion at the tip exposed. The small exposed portion (500 μm-1 mm) readily penetrated the brain allowing for an insertion that did not require the handling of each fiber by forceps. The second method involved the fabrication of silicon support structures with individual shanks spaced 150 μm apart. Each shank consisted of a small groove that held an individual carbon fiber. Our results showed that the PEG coating allowed for the chronic implantation of carbon fiber arrays in five rats with unit activity detected at 31 days post-implant. The silicon support structures recorded single unit activity in three acute rat surgeries. In one of those surgeries a stacked device with three layers of silicon support structures and carbon fibers was built and shown to readily insert into the brain with unit activity on select sites. From these studies we have found that carbon fibers spaced at ∼150 μm readily insert into the brain. This greatly increases the recording density of chronic neural probes and paves the way for even higher density devices that have a minimal scarring response.


Bystander and delayed effects after fractionated radiation exposure.
Human immortalized keratinocytes were exposed to a range of single or fractionated doses of gamma rays from (60)Co, to medium harvested from donor cells exposed to these protocols, or to a combination of radiation and irradiated cell conditioned medium (ICCM). The surviving fractions after direct irradiation or exposure to ICCM were determined using a clonogenic assay. The results show that medium harvested from cultures receiving fractionated irradiation gave lower "recovery factors" than direct fractionated irradiation, where normal split-dose recovery occurred. The recovery factor is defined here as the surviving fraction of the cells receiving two doses (direct or ICCM) separated by an interval of 2 h divided by the surviving fraction of cells receiving the same dose in one exposure. After treatment with ICCM, the recovery factors were less than 1 over a range of total doses from 5 mGy-5 Gy. Varying the time between doses from 10 min to 180 min did not alter the effect of ICCM, suggesting that two exposures to ICCM are more toxic than one irrespective of the dose used to generate the response. In certain protocols using mixtures of direct irradiation and ICCM, it was possible to eliminate the bystander effect. If bystander factors are produced in vivo, then they may reduce the sparing effect of the dose fractionation.


Sudden death due to troponin T mutations.
This study was designed to verify initial observations of the clinical and prognostic features of hypertrophic cardiomyopathy caused by cardiac tropnin T gene mutations. The most common cause of sudden cardiac death in the young is hypertrophic cardiomyopathy, which is usually familial. Mutations causing familial hypertrophic cardiomyopathy have been identified in a number of contractile protein genes, raising the possibility of genetic screening for subjects at risk. A previous report suggested that mutations in the cardiac troponin T gene were notable because they were associated with a particularly poor prognosis but only mild hypertrophy. Given the variability of some genotype:phenotype correlations, further analysis of cardiac troponin T mutations has been a priority. Deoxyribonucleic acid from subjects with hypertrophic cardiomyopathy was screened for cardiac troponin T mutations using a ribonuclease protection assay. Polymerase chain reaction-based detection of a novel mutation was used to genotype members of two affected pedigrees. Gene carriers were examined by echocardiography and electrocardiology, and a family history was obtained. A novel cardiac troponin T gene mutation, arginine 92 tryptophan, was identified in 19 of 48 members of two affected pedigrees. The clinical phenotype was characterized by minimal hypertrophy (mean [+/-SD] maximal ventricular wall thickness 11.3 +/- 5.4 mm) and low disease penetrance by clinical criteria (40% by echocardiography) but a high incidence of sudden cardiac death (mean age 17 +/- 9 years). These data support the observation that apparently diverse cardiac troponin T gene mutations produce a consistent disease phenotype. Because this is one of poor prognosis, despite deceptively mild or undetectable hypertrophy, genotyping at this locus may be particularly informative in patient management and counselling.


Facilitating peer based learning through summative assessment - An adaptation of the Objective Structured Clinical Assessment tool for the blended learning environment.
Adapting a course from face to face to blended delivery necessitates that assessments are modified accordingly. In Australia the Objective Structured Clinical Assessment tool, as a derivative from the Objective Structured Clinical Examination, has been used in the face-to-face delivery mode as a formative or summative assessment tool in medicine and nursing since 1990. The Objective Structured Clinical Assessment has been used at Charles Darwin University to assess nursing students' simulated clinical skills prior to the commencement of their clinical placements since 2008. Although the majority of the course is delivered online, students attend a one-week intensive clinical simulation block yearly, prior to attending clinical placements. Initially, the Objective Structured Clinical Assessment was introduced as a lecturer assessed summative assessment, over time it was adapted to better suit the blended learning environment. The modification of the tool from an academic to peer assessed assessment tool, was based on the empirical literature, student feedback and a cross-sectional, qualitative study exploring academics' perceptions of the Objective Structured Clinical Assessment (Bouchoucha et al., 2013a, b). This paper presents an overview of the process leading to the successful adaptation of the Objective Structured Clinical Assessment to suit the requirements of a preregistration nursing course delivered through blended learning. This is significant as many universities are moving their curriculum to fully online or blended delivery, yet little attention has been paid to adapting the assessment of simulated clinical skills. The aim is to identify the benefits and drawbacks of using the peer assessed Objective Structured Clinical Assessment and share recommendations for successful implementation.


National variation of technical factors in computed tomography of the head.
The objectives of this study were to describe head computed tomography (CT) technical factors used in the United States; assess guideline compliance and variability by sex, geographic region, institution, and manufacturer; and compare with NEXT (Nationwide Evaluation of X-ray Trends) survey. Two thousand thirty-five head CT studies from 895 sites were analyzed. Peak kilovoltage, exposure, slice and image number, slice thickness, field of view, interslice gap, reformations, examination inclusiveness, use of contrast, and reconstruction filter were compared by sex, geographic region, institution type, and manufacturer. One hundred twenty to 140 kVp (peak kilovoltage) was used by 99.8% of the sites. There was no correlation between mAs and kVp. There was a 30-fold increase in number of images and nearly 9-fold increase in number of slices from least to most. Nearly 4% use slice thickness and interslice gap greater than guidelines. There were significant regional differences in field of view, image number, and slice thickness. Some variation related to manufacturers was found. Minor difference was also found between academic and nonacademic institutions. There were significant differences in kVp and mAs compared with NEXT. Significant variations in head CT technique exist in the United States.


Microparticle movements in optical funnels and pods.
Three-dimensional microparticle movements induced by laser beams with a funnel- and tubular pod-like structure, in the neighbourhood of the focal plane of an optical trapping setup, are experimentally studied. The funnel and pod beams constructed as coherent superpositions of helical Laguerre-Gaussian modes are synthesized by a computer generated hologram using a phase-only spatial light modulator. Particle tracking is achieved by in-line holography method which allows an accurate position measurement. It is experimentally demonstrated that the trapped particle follows different trajectories depending on the orbital angular momentum density of the beam. In particular applying the proposed pod beam the particle rotates in opposite directions during its movement in the optical trap. Possible applications of these single-beam traps for volumetric optical particle manipulation are discussed.


Lactic acid bacteria and yeasts involved in the fermentation ofamabere amaruranu, a Kenyan fermented milk.
Indigenous fermented milk products contain microbiota composed of technologically important species and strains which are gradually getting lost with new technologies. We investigated the microbial diversity inamabere amaruranu, a traditionally fermented milk product from Kenya. Sixteen samples of the product from different containers were obtained. One hundred and twenty isolates of lactic acid bacteria (LAB) and 67 strains of yeasts were identified using API 50 CH and API 20 C AUX identification kits, respectively. The average pH of all the traditional fermented samples was 4.00 ± 0.93. Lactobacilli, yeasts, and molds as well asEnterobacteriaceae counts from the plastic containers were significantly higher (P < 0.05) than those from gourd.Enterobacteriaceae were below 1.00 ± 1.11 log10 cfu/mL in products from the gourds and 2.17 ± 1.92 log10 cfu/mL from the plastic containers. The LAB species were identified asStreptococcus thermophilus (25%),Lactobacillus plantarum (20%), andLeuconostoc mesenteroides (20%). The predominant yeasts wereSaccharomyces cerevisiae (25%),Trichosporum mucoides (15%),Candida famata (10%), andCandida albicans (10%). The type of vessel used for fermentation had no significant influence on the type of isolated and identified species. The diverse mixture of LAB and yeasts microflora forms a potential consortium for further product innovation inamabere amaruranu and other fermented milk products.


The efficacy of L. (L.) chagasi excreted-secreted antigens (ESAs) for visceral leishmaniasis diagnosis is due to low levels of cross-reactivity.
The analysis of promastigote excreted-secreted antigen (ESA) reactivity with 53 visceral leishmaniasis (VL) cases showed that each sample reacted regardless of the antigen or the Leishmania species used in enzyme-linked immunosorbent assay (ELISA) displayed 100% positivity with the L. (L.) chagasi ESA-blot recognizing bands of molecular weight ranging from 26.5 to 31.5 kDa. The analysis of 160 non-visceral cases showed that 5% of the samples cross-reacted with the L. (L.) chagasi ESA-ELISA and 9.4% reacted with the ESA isolated from L. (L.) amazonensis and L. (V.) braziliensis, whereas a high cross-reaction ranging from 24.4% to 25% was observed with total crude promastigote antigens (PRO-ELISA). The ESA-blot of L. (L.) chagasi tested with non-visceral sera samples showed a cross-reaction with 8.8% of cases; most of these cases represented tegumentary leishmaniasis and only one acute chagasic case. These data lead us to recommend the use of ESA as an alternative antigen in VL diagnosis.


Gemella morbillorum as the Culprit Organism of Post-Colonoscopy Necrotizing Perineal Soft Tissue Infection in a Diabetic Patient With Crohn's Disease.
Gemellsa morbillorum (G. morbillorum) is a Gram-positive facultative anaerobe and a known commensal organism of the oropharyngeal and gastrointestinal tracts. It is considered a rare cause of infections in humans. Most of the documented infections, whereas G. morbillorum has been implicated as a causative pathogen, were infective endocarditis and deep visceral abscesses. However, there are only a handful of cases in the current literature that have reported G. morbillorum as the primary organism causing necrotizing soft tissue infections. The authors presented a rare case of post-colonoscopy necrotizing perineal soft tissue infections in an elderly patient with poorly controlled diabetes mellitus and Crohn's disease with G. morbillorum being the culprit pathogen of this necrotizing infection. The reported case raises concerns for this commensal organism as an emerging virulent pathogen in certain high-risk patients. The authors proposed that a combination of the long-standing Crohn's disease and the recent colonoscopy with rectal polypectomy has predisposed the patient to G. morbillorum bacteremia with perineal sepsis in the setting of diabetic immunosuppression. Further studies are warranted to ascertain whether G. morbillorum is acquiring increased virulence that would have enabled this organism to cause novel soft tissue infections.


An assessment of implications of adaptive licensing for pharmaceutical intellectual property and regulatory exclusivity rights in the European Union.
One of the key advantages of adaptive licensing (AL) is to align the licensing of new medicines more closely with patient needs for earlier access to beneficial treatments. From an innovators perspective, "earlier" market access may seem an obvious incentive to gain earlier revenue generation. However, this is offset with an "earlier" start to patent and regulatory protection periods, which, depending on the technology, disease, population, and timing of subsequent asset protection periods, can present a conflict.


Home mechanical ventilation through mask: monitoring leakage and nocturnal oxygenation at home.
Leakage is common in patients receiving home mechanical ventilation (HMV) via a face mask. Although pressure ventilators have partial compensatory capacity, excessive leakage can compromise the effectiveness of treatment. Home ventilators are equipped with built-in software which provides information on leakage. However, the values of leakage and their effects in routine clinical practice are currently little known. To measure leakage in stable patients on nocturnal HMV and its impact on treatment effectiveness. Consecutive outpatients on HMV were recruited. Nocturnal pulse oximetry was performed at home and leakage was measured using the ventilator's built-in software. We measured: mean SpO(2), percentage of time with SpO(2) <90% (T90), mean leakage (meanL), maximum leakage (maxL), and minimum leakage (minL) during the ventilation session. We estimated ventilator capacity to compensate for leakage according to inspiratory positive airway pressure and divided the patients into two groups: those with leak compensation and those without. The study included 41 patients [mean age, 64 years (SD 11.9); 23 (56%) women]. Nocturnal pulse oximetry showed an SpO(2) of 94% (±2.9) and a T90 of 10% (±21.7). Leakage (in l/min) was: meanL, 32.2 (±15.3); maxL, 64.8 (±28.5), and minL, 18.8 (±10.6). Seven cases (17%) had leakage greater than the ventilator compensatory capacity, but no significant difference in SpO(2) or T90 was observed between patients with or without leak compensation. A wide variation between maxL and minL was observed in our series; 17% of cases had higher leakage values than the compensatory capacity of the ventilator, but this did not affect nocturnal oxygenation.


The dynamics of grooming interactions: maintenance of partner choice and the consequences of demographic variation for female mandrills.
A large body of evidence suggests that female Old World monkeys maintain selective long-term grooming interactions with fitness benefits. The last two decades have produced evidence that the regulation of social interactions among primates can be, in part, explained by the Biological Markets theory, with grooming behaviour as the focus of these studies. Grooming facilitates bonding between individuals, constituting an essential part of the regulation of social relationships among female cercopithecids. In contrast to the well-studied baboons (Papio spp), knowledge about the nature of grooming interactions and their regulation is generally lacking for the large, terrestrial species of mandrills (Mandrillus sphinx). We used a combination of social network analysis tools and well-established methods for assessing partner diversity and reciprocity to characterise grooming networks, partner choice and patterns of trade (be groomed, give grooming) among females in a captive group of mandrills, both within and across two separate observation periods. Our results suggest that, even though the relatively stable conditions of captivity allowed the studied females to maintain selective grooming interactions across time, small scale demographic changes affected the grooming dynamics of the group in accordance with the expectations of the Biological Markets theory. In particular, the maturation and consequent integration of a high ranking female into the group's grooming network from one period to the next resulted in a more pronounced effect of rank on the regulation of grooming interactions. In addition, the influence of the maturation of a dependent infant on the grooming interactions of his mother were evident between periods. Our results also demonstrate that grooming networks are dynamic and that high ranking individuals are not necessarily the most central in grooming networks. Finally, we discuss the potential of social network analysis to identify cases of social exclusion and its consequences for captive management.


Medical and pharmacy coverage decision making at the population level.
Medicare is one of the largest health care payers in the United States. As a result, its decisions about coverage have profound implications for patient access to care. In this commentary, the authors describe how Medicare used evidence on heterogeneity of treatment effects to make population-based decisions on health care coverage for implantable cardiac defibrillators. This case is discussed in the context of the rapidly expanding availability of comparative effectiveness research. While there is a potential tension between population-based and patient-centered decision making, the expanded diversity of populations and settings included in comparative effectiveness research can provide useful information for making more discerning and informed policy and clinical decisions.


Studies of the mechanism of cytolysis by HIV-1-specific CD4+ human CTL clones induced by candidate AIDS vaccines.
Vaccine-induced, virus-specific CTLs may rapidly eliminate the host cells that first become infected after virus exposure, thereby preventing disseminated infection. Thus, there is much interest in the ability of candidate AIDS vaccines to elicit CTLs. All HIV-1 envelope (env) protein-based vaccines tested to date in seronegative humans induce CTLs from the CD4+ subset. Because the mechanism of cytolysis by CD4+ CTLs is controversial, a detailed study of the cytolytic reactions mediated by vaccine-induced, HIV-1-specific human CD4+ CTL clones was conducted. CD4+ CTL clones induced rapid destruction of Ag-pulsed target cells. Lysis was readily detectable within 15 min. Lysis was not a result of syncytium formation between CD4+ effector cells and env-expressing targets. Target cell destruction was not dependent upon de novo RNA or protein synthesis in either the effector or the target cell. Expression of perforin mRNA was detected by Northern blotting and reverse-transcriptase-PCR in CD4+ CTL clones but not in autologous B lymphoblastoid cell lines. Immunohistochemical studies demonstrated perforin protein in cytoplasmic granules in CD4+ CTL clones. Lysis by CD4+ CTLs was strictly dependent upon extracellular Ca2+ and was highly specific, with no lysis of innocent bystander cells. DNA fragmentation was detectable in target cells, but did not precede 51Cr release. Taken together, these results provide a dramatically different view of cytolysis by human CD4+ CTLs. Target cells are lysed by a rapid and efficient mechanism that involves a preformed mediator and that is functionally similar to the mechanism used by CD8+ CTLs.


Stem cells for lupus nephritis: a concise review of current knowledge.
Lupus nephritis (LN), a common manifestation of systemic lupus erythematosus (SLE), accounts for significant morbidity and mortality in SLE patients. Since the available standard therapies and biologic agents for LN are yet to achieve the desired response and have considerable secondary effects, stem cell therapy has now emerged as a new approach. This therapy involves the transplantation of hematopoietic stem cells (HSCs) and mesenchymal stem cells (MSCs). Our current review will highlight the progress of stem cell therapy for LN, along with the challenges encountered and the future direction of this approach.


Collagen vitrigel: a novel scaffold that can facilitate a three-dimensional culture for reconstructing organoids.
Three-dimensional reconstructed organoids in vitro are valuable for not only regenerative medicine but also drug development. However, the manipulation of conventional three-dimensional cultures is not simple. We describe a nylon membrane ring-embedded or a pressed silk sheet-embedded scaffold made of collagen "vitrigel" that can facilitate three-dimensional cultures for reconstructing an epithelial-mesenchymal model or a hard connective tissue model, respectively. Here we define vitrigel as a gel in a stable state produced by rehydration after the vitrification of a traditional hydrogel. The collagen vitrigel was successfully prepared in three steps involving a gelation process in which a cold and clear neutral salt solution containing type I collagen formed an opaque and soft gel by incubation at 37 degrees C, a vitrification process in which the gel becomes a rigid material like glass after sufficient drying out, and finally a rehydration process to convert the vitrified material into a thin and transparent gel membrane with enhanced gel strength. The framework-embedded collagen vitrigel scaffold that can be easily reversed by forceps was prepared by inserting a nylon ring or a silk sheet in the collagen solution prior to the gelation. The scaffold enabled culturing anchorage-dependent cells on both surfaces of the collagen vitrigel by the manipulation of two-dimensional cultures and consequently resulted in reconstructing a three-dimensional organoid. An intestinal epithelial-mesenchymal model was reconstructed by coculturing fibroblasts on the opposite side of monolayered Caco-2 cells on the nylon ring-embedded collagen vitrigel. Also, fibroblasts seeded on both surfaces of the silk sheet-embedded collagen vitrigel proliferated well and formed multilayers and some cells invaded into the vitrigel framed by the network of numerous strong silk filaments, suggesting a reconstruction of a hard connective tissue model. These data demonstrate that the collagen vitrigel is a valuable scaffold for tissue engineering.


Aggressive B-cell Lymphoma with MYC/TP53 Dual Alterations Displays Distinct Clinicopathobiological Features and Response to Novel Targeted Agents.
Diffuse large B-cell lymphoma (DLBCL) is the major type of aggressive B-cell lymphoma. High-grade B-cell lymphoma (HGBCL) with MYC/BCL2 double-hit (DH) represents a distinct entity with dismal prognosis after standard immunochemotherapy in the current WHO lymphoma classification. However, whether TP53 mutation synergizes with MYC abnormalities (MYC rearrangement and/or Myc protein overexpression) contributing to HGBCL-like biology and prognosis is not well investigated. In this study, patients with DLBCL with MYC/TP53 abnormalities demonstrated poor clinical outcome, high-grade morphology, and distinct gene expression signatures. To identify more effective therapies for this distinctive DLBCL subset, novel MYC/TP53/BCL-2-targeted agents were investigated in DLBCL cells with MYC/TP53 dual alterations or HGBCL-MYC/BCL2-DH. A BET inhibitor INCB057643 effectively inhibited cell viability and induced apoptosis in DLBCL/HGBCL cells regardless of MYC/BCL2/TP53 status. Combining INCB057643 with a MDM2-p53 inhibitor DS3032b significantly enhanced the cytotoxic effects in HGBCL-DH without TP53 mutation, while combining with the BCL-2 inhibitor venetoclax displayed potent therapeutic synergy in DLBCL/HGBCL cells with and without concurrent TP53 mutation. Reverse-phase protein arrays revealed the synergistic molecular actions by INCB057643, DS3032b and venetoclax to induce cell-cycle arrest and apoptosis and to inhibit AKT/MEK/ERK/mTOR pathways, as well as potential drug resistance mechanisms mediated by upregulation of Mcl-1 and RAS/RAF/MEK/ERK pathways. In summary, these findings support subclassification of DLBCL/HGBCL with dual MYC/TP53 alterations, which demonstrates distinct pathobiologic features and dismal survival with standard therapy, therefore requiring additional targeted therapies. IMPLICATIONS: The clinical and pharmacologic studies suggest recognizing DLBCL with concomitant TP53 mutation and MYC abnormalities as a distinctive entity necessary for precision oncology practice. VISUAL OVERVIEW: http://mcr.aacrjournals.org/content/molcanres/19/2/249/F1.large.jpg.


Aromatase inhibitors and musculoskeletal pain in patients with breast cancer.
Aromatase inhibitors are recommended for use by postmenopausal women who have estrogen receptor-positive early-stage breast cancer. They reduce local and distant recurrence more effectively than tamoxifen. Anastrozole (Arimidex, AstraZeneca Pharmaceuticals LP), letrozole (Femara, Novartis Pharmaceuticals Corporation), and exemestane (Aromasin, Pfizer Inc.) inhibit aromatase activity, thus significantly decreasing estrogen production in tissues such as liver, muscle, and fat. Very low levels of estrogen may be one cause of musculoskeletal pain, a common side effect associated with the drugs. In the major adjuvant aromatase inhibitor clinical trials, 25%-30% of the patients enrolled experienced musculoskeletal pain. Although quality-of-life studies demonstrate that aromatase inhibitors are well tolerated overall, some women discontinue this treatment because of musculoskeletal pain. Little is known about how to predict, measure, or manage musculoskeletal pain caused by aromatase inhibition. Oncology nurses play an important role in the assessment and management of side effects related to cancer. This article provides an overview of the current knowledge about musculoskeletal pain in patients with breast cancer receiving aromatase inhibitor therapy.


Trafficking of Kv2.1 Channels to the Axon Initial Segment by a Novel Nonconventional Secretory Pathway.
Kv2.1 is a major delayed-rectifier voltage-gated potassium channel widely expressed in neurons of the CNS. Kv2.1 localizes in high-density cell-surface clusters in the soma and proximal dendrites as well as in the axon initial segment (AIS). Given the crucial roles of both of these compartments in integrating signal input and then generating output, this localization of Kv2.1 is ideal for regulating the overall excitability of neurons. Here we used fluorescence recovery after photobleaching imaging, mutagenesis, and pharmacological interventions to investigate the molecular mechanisms that control the localization of Kv2.1 in these two different membrane compartments in cultured rat hippocampal neurons of mixed sex. Our data uncover a unique ability of Kv2.1 channels to use two molecularly distinct trafficking pathways to accomplish this. Somatodendritic Kv2.1 channels are targeted by the conventional secretory pathway, whereas axonal Kv2.1 channels are targeted by a nonconventional trafficking pathway independent of the Golgi apparatus. We further identified a new AIS trafficking motif in the C-terminus of Kv2.1, and show that putative phosphorylation sites in this region are critical for the restricted and clustered localization in the AIS. These results indicate that neurons can regulate the expression and clustering of Kv2.1 in different membrane domains independently by using two distinct localization mechanisms, which would allow neurons to precisely control local membrane excitability.SIGNIFICANCE STATEMENT Our study uncovered a novel mechanism that targets the Kv2.1 voltage-gated potassium channel to two distinct trafficking pathways and two distinct subcellular destinations: the somatodendritic plasma membrane and that of the axon initial segment. We also identified a distinct motif, including putative phosphorylation sites, that is important for the AIS localization. This raises the possibility that the destination of a channel protein can be dynamically regulated via changes in post-translational modification, which would impact the excitability of specific membrane compartments.


Downregulation of Cancer Stemness by Novel Diterpenoid Ovatodiolide Inhibits Hepatic Cancer Stem Cell-Like Traits by Repressing Wnt/[Formula: see text]-Catenin Signaling.
The hierarchical tumor propagation or cancer stem cells (CSCs) model of carcinogenesis postulates that like physiologic adult stem cell (ASC), the CSCs positioned at the apex of any tumor population form the crux of tumor evolution with a constitutive regenerative capacity and differentiation potential. The propagation and recurrence of the characteristically heterogeneous and therapy-resistant hepatocellular carcinoma (HCC), adds to accumulating evidence to support this CSCs model. Based on the multi-etiologic basis of HCC formation which among others, focuses on the disruption of the canonical Wnt signaling pathway, this study evaluated the role of cembrane-type phytochemical, Ovatodiolide, in the modulation of the Wnt/[Formula: see text]-catenin pathway, and its subsequent effect on liver CSCs' activities. Our fluorescence-activated cell sorting (FACS) and quantitative RT-PCR analyses of side population (SP) indicated that CD133+ cells were [Formula: see text]-catenin-overexpressing, more aggressive, and resistant to the conventional anticancer agents, Cisplatin and Doxorubicin, when compared to [Formula: see text]-catenin-downregulated group. We demonstrated that marked upregulation of [Formula: see text]-catenin and its downstream targets effectively enhanced hepatosphere formation, with an associated induction of CD133, OCT4 and Sox2 expression and also caused an significant enhancement of HCC proliferation. However, treatment with Ovatodiolide induced downregulation of [Formula: see text]-catenin and its downstream effector genes, abolished hepatosphere formation and reversed the [Formula: see text]-catenin-associated enhancement of HCC growth. In summary, we demonstrated for the first time that Ovatodiolide suppressed the canonical Wnt signaling pathway, and inhibited the generation of liver CSCs; Thus, projecting Ovatodiolide as a putatively effective therapeutic agent for anti-HCC target therapy.


Altered profile of secondary metabolites in the root exudates of Arabidopsis ATP-binding cassette transporter mutants.
Following recent indirect evidence suggesting a role for ATP-binding cassette (ABC) transporters in root exudation of phytochemicals, we identified 25 ABC transporter genes highly expressed in the root cells most likely to be involved in secretion processes. Of these 25 genes, we also selected six full-length ABC transporters and a half-size transporter for in-depth molecular and biochemical analyses. We compared the exuded root phytochemical profiles of these seven ABC transporter mutants to those of the wild type. There were three nonpolar phytochemicals missing in various ABC transporter mutants compared to the wild type when the samples were analyzed by high-performance liquid chromatography-mass spectrometry. These data suggest that more than one ABC transporter can be involved in the secretion of a given phytochemical and that a transporter can be involved in the secretion of more than one secondary metabolite. The primary and secondary metabolites present in the root exudates of the mutants were also analyzed by gas chromatography-mass spectrometry, which allowed for the identification of groups of compounds differentially found in some of the mutants compared to the wild type. For instance, the mutant Atpdr6 secreted a lower level of organic acids and Atmrp2 secreted a higher level of amino acids as compared to the wild type. We conclude that the release of phytochemicals by roots is partially controlled by ABC transporters.


The Effects of Bilirubin and Lumirubin on the Differentiation of Human Pluripotent Cell-Derived Neural Stem Cells.
The 'gold standard' treatment of severe neonatal jaundice is phototherapy with blue-green light, which produces more polar photo-oxidation products that are easily excreted via the bile or urine. The aim of this study was to compare the effects of bilirubin (BR) and its major photo-oxidation product lumirubin (LR) on the proliferation, differentiation, morphology, and specific gene and protein expressions of self-renewing human pluripotent stem cell-derived neural stem cells (NSC). Neither BR nor LR in biologically relevant concentrations (12.5 and 25 µmol/L) affected cell proliferation or the cell cycle phases of NSC. Although none of these pigments affected terminal differentiation to neurons and astrocytes, when compared to LR, BR exerted a dose-dependent cytotoxicity on self-renewing NSC. In contrast, LR had a substantial effect on the morphology of the NSC, inducing them to form highly polar rosette-like structures associated with the redistribution of specific cellular proteins (β-catenin/N-cadherin) responsible for membrane polarity. This observation was accompanied by lower expressions of NSC-specific proteins (such as SOX1, NR2F2, or PAX6) together with the upregulation of phospho-ERK. Collectively, the data indicated that both BR and LR affect early human neurodevelopment in vitro, which may have clinical relevance in phototherapy-treated hyperbilirubinemic neonates.


A single-bout of one-hour spinning exercise increases troponin T in healthy subjects.
While long-term endurance exercise is known to increase cardiac biomarkers, only a few studies on short-term exercise and these markers have been reported. The aim of this study was to investigate the acute effects of a one-hour bicycle spinning on cardiac biomarkers in healthy individuals. Serum levels of high-sensitive troponin T (TnT), creatinine kinase MB fraction (CK-MB), N-terminal pro-brain natriuretic peptide (NT-proBNP), creatinine kinase (CK) and myoglobin were measured at baseline, 1 and 24 hour after one hour of spinning exercise in ten healthy and fit (age 31.0 ± 6.6 years) individuals. TnT doubled one hour post-exercise (All values ≤ 5 - 9.7 ± 6.0 ng/L, p < 0.001). Two individuals had TnT levels above upper reference limit, URL (20.7 and 20.2 ng/L, URL = 12 ng/L). Myoglobin levels increased 72% one hour post-exercise (38 ± 20 - 66 ± 41 mg/L, p < 0.02). TnT and myoglobin levels returned to baseline 24 hour post-exercise. Serum levels of CK-MB, NT-proBNP and CK were not significantly changed. A single-bout of one-hour bicycle spinning transiently increases TnT and myoglobin in healthy subjects. Some subjects even have TnT release above URL. Thus, recently performed exercise also of short duration should be taken into consideration in the evaluation of acute chest pain with release of cardiac TnT.


Material chemistry of graphene oxide-based nanocomposites for theranostic nanomedicine.
The promising biomedical applications of graphene oxide (GO) have quickly catalyzed a generation of various GO-based functional composites with desirable nanostructures and composites to satisfy the specific biomedical applications of GO-based nanosystems. This comprehensive review summarizes and discusses the very recent developments in rational design, chemical fabrication and versatile biomedical applications of GO-based functional composites, including metal oxide-GO, metal-GO, fluorescent nanoparticles-GO, silica-GO, polymer-GO and other specific GO-based composite nanosystems. The integration of multi-components with GO is also discussed. Their versatile theranostic applications include diagnostic imaging-guided therapy (photothermal therapy, photodynamic therapy and chemotherapy), biosensing-based tumor detection, and synergistic therapy. The biocompatibility and biosafety of these promising GO-based nanocomposites are also briefly mentioned. Finally, the unresolved critical issues and challenges facing the design/fabrication/application of these unique GO-based functional nanocomposites are proposed to further promote their clinical translation. The unique multi-composition, nanostructure and unique theranostic performance of GO-based composites are highly expected to pave a new way for the development of highly efficient theranostic protocols/approaches for precise personalized biomedicine to benefit human health.


Invasive vegetation affects amphibian skin microbiota and body condition.
Invasive plants are major drivers of habitat modification and the scale of their impact is increasing globally as anthropogenic activities facilitate their spread. In California, an invasive plant genus of great concern is Eucalyptus. Eucalyptus leaves can alter soil chemistry and negatively affect underground macro- and microbial communities. Amphibians serve as excellent models to evaluate the effect of Eucalyptus invasion on ground-dwelling species as they predate on soil arthropods and incorporate soil microbes into their microbiotas. The skin microbiota is particularly important to amphibian health, suggesting that invasive plant species could ultimately affect amphibian populations. To investigate the potential for invasive vegetation to induce changes in microbial communities, we sampled microbial communities in the soil and on the skin of local amphibians. Specifically, we compared Batrachoseps attenuatus skin microbiomes in both Eucalyptus globulus (Myrtaceae) and native Quercus agriflolia (Fagaceae) dominated forests in the San Francisco Bay Area. We determined whether changes in microbial diversity and composition in both soil and Batrachoseps attenuatus skin were associated with dominant vegetation type. To evaluate animal health across vegetation types, we compared Batrachoseps attenuatus body condition and the presence/absence of the amphibian skin pathogen Batrachochytrium dendrobatidis. We found that Eucalyptus invasion had no measurable effect on soil microbial community diversity and a relatively small effect (compared to the effect of site identity) on community structure in the microhabitats sampled. In contrast, our results show that Batrachoseps attenuatus skin microbiota diversity was greater in Quercus dominated habitats. One amplicon sequence variant identified in the family Chlamydiaceae was observed in higher relative abundance among salamanders sampled in Eucalyptus dominated habitats. We also observed that Batrachoseps attenuatus body condition was higher in Quercus dominated habitats. Incidence of Batrachochytrium dendrobatidis across all individuals was very low (only one Batrachochytrium dendrobatidis positive individual). The effect on body condition demonstrates that although Eucalyptus may not always decrease amphibian abundance or diversity, it can potentially have cryptic negative effects. Our findings prompt further work to determine the mechanisms that lead to changes in the health and microbiome of native species post-plant invasion.


Impact of hypertension stratified by diabetes on the lifetime risk of cardiovascular disease mortality in Japan: a pooled analysis of data from the Evidence for Cardiovascular Prevention from Observational Cohorts in Japan study.
Lifetime risk is an informative estimate to motivate people to change lifestyle behaviors, especially from a younger age, in public health education. The impact of the combination of hypertension and diabetes on the lifetime risk of cardiovascular mortality has not been investigated in Asian populations. A pooled analysis of individual data from nine cohorts was performed. A total of 57,339 Japanese men and women were eligible for the analysis. We used the modified Kaplan-Meier approach and estimated the remaining lifetime risk of cardiovascular mortality starting from the index age of 35 years. Participants were classified into four categories defined by hypertension and diabetes. The lifetime risk was increased in the order of those without either risk, those without hypertension but with diabetes, those with hypertension but without diabetes, and those with both risks. The lifetime risk of cardiovascular mortality at the 35-year index age was as follows: 7.8% in men and 6.2% in women for those without either hypertension or diabetes, 13.2% in men and 9.5% in women for those without hypertension but with diabetes, 17.2% in men and 11.7% in women for those with hypertension but without diabetes, and 19.4% in men and 15% in women for those with both risks. These findings reinforce the need for a life-course perspective in the management of hypertension and diabetes from a younger age.


In Vitro Production of Radiolabeled Red Clover (Trifolium pratense) Isoflavones.
Red clover isoflavones are increasingly used in dietary supplements for their purported estrogenic effects. However, little is known about their metabolism in animals due to a lack of commercially available isotopically-labeled tracers. The goal of this research was to establish red clover cell culturing methodology for (14)C-biolabeling of isoflavones. When root, leaf, and petiole-derived suspension cultures were grown in darkness or light, dark-grown, petiole-derived solution cultures produced the highest concentrations of the two major red clover isoflavones, formononetin (0.67 mg/g FM inoculum) and biochanin A (0.13 mg/g FM inoculum). Varying levels and timing of copper chloride elicitor did not significantly affect isoflavone accumulation. Approximately 38% of the (14)C-sucrose dose accumulated in the cells. Eighteen percent of the initial labeled dose was detected in the isoflavone-rich methanolic extract and of that, 22% accumulated in isoflavones.


Communication: an efficient analytic gradient theory for approximate spin projection methods.
Spin polarized and broken symmetry density functional theory are popular approaches for treating the electronic structure of open shell systems. However, spin contamination can significantly affect the quality of predicted geometries and properties. One scheme for addressing this concern in studies involving broken-symmetry states is the approximate projection method developed by Yamaguchi and co-workers. Critical to the exploration of potential energy surfaces and the study of properties using this method will be an efficient analytic gradient theory. This communication introduces such a theory formulated, for the first time, within the framework of general post-self consistent field (SCF) derivative theory. Importantly, the approach taken here avoids the need to explicitly solve for molecular orbital derivatives of each nuclear displacement perturbation, as has been used in a recent implementation. Instead, the well-known z-vector scheme is employed and only one SCF response equation is required.


Central Venous Catheterization: Are We Using Ultrasound Guidance?
To assess the self-reported frequency of use of ultrasound guidance for central venous catheterization by emergency medicine (EM) residents, describe residents' perceptions regarding the use of ultrasound guidance, and identify barriers to the use of ultrasound guidance. A longitudinal cross-sectional study was conducted at 5 academic institutions. A questionnaire on the use of ultrasound guidance for central venous catheterization was initially administered to EM residents in 2007. The same questionnaire was distributed again in the 5 EM residency programs in 2013. In 2007 and 2013, 147 and 131 residents completed questionnaires, respectively. A significant increase in the use of ultrasound guidance for central venous catheterization was reported in 2013 compared to 2007 (P< .001). In 2007, 53% (95% confidence interval, 44%-61%) of residents reported that they were initially trained in central venous catheterization using ultrasound guidance compared to 96% (95% confidence interval, 92%-99%) in 2013 (P < .0001). In 2007, more residents thought that faculty were insufficiently adopting ultrasound (42% versus 9%), and there was a lack of ultrasound teaching during residency training (14% versus 5%) compared to 2013. The use of self-reported ultrasound guidance for central venous catheterization significantly increased from 2007 to 2013 at academic institutions. Most residents were aware of the benefits of using ultrasound guidance. Although faculty adoption of ultrasound for central venous catheterization remains a barrier, it has decreased.


Enhanced marrow adipogenesis and bone resorption in estrogen-deprived rats treated with the PPARgamma agonist BRL49653 (rosiglitazone).
Thiazolidinediones are insulin-sensitizing agents and in clinical use for the treatment of type II diabetes. Under specific experimental conditions, these molecules induce adipogenic differentiation of mesenchymal precursor cells at the expense of osteoblasts in vitro, suggesting possible negative effects on the skeleton. We measured effects of the thiazolidinedione BRL49653 on bone tissue of intact and estrogen-deprived skeletally mature adult female Wistar rats (6-9 months old). Weight gain and decreased plasma triglyceride levels confirmed the effectiveness of the treatment. However, no change in bone mass or fat marrow volume was observed in intact rats treated for 8 weeks with 5, 10, or 20 mg/kg of BRL49653. Study of marrow cultures established at necropsy revealed a higher responsiveness to adipogenic differentiation protocols of cultures established from the 10-mg/kg group compared to vehicle control. In a second study, the effects of thiazolidinedione treatment on the skeleton of estrogen-deprived rats were investigated. Application of 10 mg/kg of BRL49653 for 12 weeks resulted in enhanced bone loss (+31%; pQCT) and increased fat marrow volume (+117%; histomorphometry) compared to vehicle-treated OVX control. Interestingly, osteoblast number was comparable in both cases. Bone resorption parameters were significantly increased in the treatment group (+27% osteoclast number, +30% eroded surface). Enhanced bone loss due to treatment was consistently observed in the tibia, femur, and the lumbar spine. Our data indicate that thiazolidinediones may enhance bone loss induced by estrogen deprivation.