Structural determinants of substrates for the prostaglandin transporter PGT.
We recently identified a broadly expressed transporter, PGT, that transports primarily prostaglandins E2 and F2 alpha (PGE2 and PGF2 alpha). In the current study, we examined the structural determinants of potential PGT substrates in detail. Rat PGT was transiently expressed in HeLa cells, the timed uptake of tracer PGE2 was determined in the presence of various concentrations of unlabeled prostanoids; and the resulting inhibitory constants (Ki) were determined by curve-fitting. PGE2 and PGF2 alpha, both known to be transported, had similar affinities for PGT (Ki = 49-50 nM). The strongest interaction (Ki = 13-19 nM) was obtained with prostanoids lacking the 9- or 11-position oxygen groups. A relatively high affinity was also obtained for the bicycloendoperoxides U44069, PGH2, and U46619 (Ki = 29-39 nM). However, a radioactive representative from this group, U46619, was not transported. Structural modifications that produced a moderately reduced affinity relative to that of PGE2 (Ki = 56-286 nM) included reduction in C5 = C6, the addition of a benzene group at position C18, and isomerization at the C8 position. In complementary studies, tracer isoprostane B-iso-PGF2 alpha was found to be transported at approximately 13% the rate of tracer PGE2. Substantially weaker interaction (Ki = > 700 nM) was seen when the 1-position COO- anionic group was neutralized or when the 15(S)-OH group was changed to 15(R)-OH or to 15-keto. These results with the cloned rat PGT are very similar to those previously reported in the in vitro perfused rat lung and indicate that PGT probably represents the predominant route by which certain prostanoids, including F2 isoprostanes, are transported across plasma membranes.


Annual Baseline King-Devick Oculomotor Function Testing Is Needed Due to Scores Varying by Age.
To document baseline King-Devick (K-D) oculomotor function scores for male and female participants aged between 4 and 20 years old. Utilising a cross section of schools, rugby clubs and gymnastic clubs, 1936 participants (1300 male, 636 female) completed the spiral-bound K-D test for the identification of disturbed oculomotor function. This study identified that overall, the baseline scores of the K-D test became faster by 1.4 (0.3 to 4.5) s per year, when compared with the previous age group in the same number of reading card groups. When comparing normative values of the original K-D validation study with the same age groups of the current cohort, participants aged 6 to 11 years recorded a faster baseline time (range 3.5 to 8.6 s), while those in the 12 to 14 years. age group recorded slower baseline times (range -3.9 to -7.9 s). In general, there were age group differences, but not sex differences, for K-D test times in the current cohort. Analysis of single card times, across all age groups, showed changes likely due to improved reading time. The results support the need for individualised annual pre-injury baseline testing of the K-D test.


Nutrition of women with hair loss problem during the period of menopause.
During the period of menopause as an effect of changes in hormone status, one of the most common ailments for women is hair loss. Taking into consideration fact that the ingredients of diet contained in various groups of consumed food products are both precursors in steroid hormones synthesis as well as have direct impact on structure, growth and keeping hair in skin integument, this is the reason why nourishing support for women during this period of life as well as during the hair loss therapy is reasonable. Standard value proteins containing Sulphur amino-acids: cysteine and methionine as precursor to keratin hair protein synthesis are basic element of diet conditioning of hair building. Irreplaceable having impact on keeping hair in skin integument is exogenous L-lysine, mainly present in the inner part of hair root is responsible for hair shape and volume. Fats present in the diet take part in steroid hormones synthesis (from cholesterol) thus have influence on keeping hair in skin integument. Women diet should contain products rich in complex carbohydrates, with low glycemic index and load containing fiber regulating carbohydrate-lipid metabolism of the body. Vitamins also have impact on the state of hair: C vitamin, group B and A vitamins. Minerals which influence hair growth are: Zn, Fe, Cu, Se, Si, Mg and Ca. It is worthwhile to pay closer attention to diet in women who besides hormone changes and undertaken pharmacotherapy are additionally exposed to chronic stress and improperly conducted cosmetic's and hairdresser's treatments.


Relationship between leukocyte telomere length and personality traits in healthy subjects.
It has been shown that certain personality traits are related to mortality and disease morbidity, but the biological mechanism linking them remains unclear. Telomeres are tandem repeat DNA sequences located at the ends of chromosomes, and shorter telomere length is a predictor of mortality and late-life disease morbidity. Thus, it is possible that personality traits influence telomere length. In the present study, we examined the relationship of leukocyte telomere length with personality traits in healthy subjects. The subjects were 209 unrelated healthy Japanese who were recruited from medical students at 4th-5th grade. Assessment of personality traits was performed by the Revised NEO Personality Inventory (NEO-PI-R) and the Temperament and Character Inventory (TCI). Leukocyte relative telomere length was determined by a quantitative real-time PCR method for a ratio of telomere/single copy gene. In the stepwise multiple regression analysis, shorter telomere length was related to lower scores of neuroticism (P<0.01) and conscientiousness (P<0.05) of the NEO-PI-R, and lower scores of harm avoidance (P<0.05) and reward dependence (P<0.05) of the TCI. The present study suggests that leukocyte telomere length is associated with some personality traits, and this association may be implicated in the relationship between personality traits and mortality.


[Periodic classification of DNA sequences recognized by restrictases: properties of symmetry and regularity].
A circular periodic map of short palindromic DNA sequences is constructed using the sequence homology and symmetry. It is applied to compare sequences recognised by class II restriction and modification enzymes with other similar DNA sequences. All known restriction sites have two strong properties: they are enriched by GC pairs, and clustered (purine-purine, pyrimidine-pyrimidine) bonds predominant upon alternating ones. The preference of AT/GC alternation is only slight. These properties were compared quantitatively with the help of suggested numerical methods among different groups of restriction enzymes. The map is applied for prediction of new specificities of restriction modification systems. Possible mechanism of DNA sequence recognition by these enzymes and their evolution are discussed.


Preliminary report of a multicentric study on environmental risk factors in Ta-T1 transitional cell carcinoma of the bladder. A study from Gruppo Studi Tumori Urologici Foundation.
The distribution of potential environmental risk factors among patients affected by superficial transitional cell carcinoma of the bladder (TCCB) has been analyzed. Patients affected by superficial TCCB underwent TUR and early intravesical chemotherapy. Detailed data about age, sex, residence, employment, active and passive cigarette smoking, water resource and hair dye use were centralized. Analysis has been conducted on 474 patients affected by Ta-T1 G1-2 TCCB at medium risk for recurrence. Patients with primary single Ta G1-2, Tis or T1G3 tumors were excluded from the present analysis. Over 80% of the patients lived in urban areas, 22% were employed in industries presumed at risk for bladder cancer, 8% used hair dye and 75% were smokers. Bottled water was the only water resource in 42% of the patients. Employment in industry at risk (p = 0.01) and cigarette smoking (p = 0.04) resulted in being statistically related to tumor multiplicity. Moreover, the period of cigarette smoking was significantly longer in patients with recurrent tumors (p = 0.026). The municipal water supply represented the main water source in never-smokers (p = 0.01) rather than in smokers and in patients harboring T1 rather than Ta tumors (p = 0.03). Employment in industry at risk and cigarette smoking resulted in being related to tumor multiplicity. The length of exposure to cigarette smoking was related to the natural history of the tumor. A drinkable water source emerged as a risk factor in absence of cigarette smoking.


Medical rehousing.
A local authority and its medical adviser collaborated to assess the needs of applicants who have special requirements for medical rehousing. Some characteristics of the applicants were examined, together with how successfully their needs were met. Over half the applicants were aged over 55. Some three years after their initial application 41% of applicants considered to have medical priority had been rehoused compared with 36% of those with no medical priority.


Enhanced understanding of infectious diseases by fusing multiple datasets: a case study on malaria in the Western Brazilian Amazon region.
A common challenge to the study of several infectious diseases consists in combining limited cross-sectional survey data, collected with a more sensitive detection method, with a more extensive (but biased) syndromic sentinel surveillance data, collected with a less sensitive method. Our article describes a novel modeling framework that overcomes this challenge, resulting in enhanced understanding of malaria in the Western Brazilian Amazon. A cohort of 486 individuals was monitored using four cross-sectional surveys, where all participants were sampled regardless of symptoms (aggressive-active case detection), resulting in 1,383 microscopy and 1,400 polymerase chain reaction tests. Data on the same individuals were also obtained from the local surveillance facility (i.e., passive and active case detection), totaling 1,694 microscopy tests. Our model accommodates these multiple pathogen and case detection methods. This model is shown to outperform logistic regression in terms of interpretability of its parameters, ability to recover the true parameter values, and predictive performance. We reveal that the main infection determinant was the extent of forest, particularly during the rainy season and in close proximity to water bodies, and participation on forest activities. We find that time residing in Acrelandia (as a proxy for past malaria exposure) decreases infection risk but surprisingly increases the likelihood of reporting symptoms once infected, possibly because non-naïve settlers are only susceptible to more virulent Plasmodium strains. We suggest that the search for asymptomatic carriers should focus on those at greater risk of being infected but lower risk of reporting symptoms once infected. The modeling framework presented here combines cross-sectional survey data and syndromic sentinel surveillance data to shed light on several aspects of malaria that are critical for public health policy. This framework can be adapted to enhance inference on infectious diseases whenever asymptomatic carriers are important and multiple datasets are available.


Uptake of colorectal cancer screening among Ontarians with intellectual and developmental disabilities.
Under-screening for cancer may contribute to a greater disease burden in individuals with intellectual and developmental disabilities (IDD) as their life expectancy increases. In 2008, the province of Ontario launched Canada's first population-based colorectal cancer screening program. Our objectives were to compare the proportions of Ontarians with and without IDD who have undergone colorectal cancer screening and to examine factors associated with screening uptake among Ontarians with IDD. Records for Ontario residents 50-64 years of age were linked across various administrative health and social services datasets to identify individuals with IDD and to select a random sample of the age-equivalent Ontario population without IDD as a comparison group. Logistic regression models were fit to examine the odds of screening uptake among individuals with IDD while controlling for age, sex, urban or rural residence, neighbourhood income quintile, expected use of health care resources, and being enrolled with or seeing a physician in a patient enrolment model (any of several primary care practice models designed to improve patient access and quality of care in Ontario), and to examine the association between these variables and colorectal cancer screening in the IDD population. The odds of having had a fecal occult blood test in the previous two years and being up-to-date with colorectal tests were 32% and 46% lower, respectively, for Ontarians with IDD compared to those without IDD. Being older, female, having a greater expected use of health care resources, and being enrolled with or seeing a physician in a primary care patient enrolment model were all significantly associated with higher odds of having been screened for colorectal cancer in the IDD population. These findings underscore the need for targeted interventions aimed at making colorectal cancer screening more equitable.


Intrinsic Expression of Immune Checkpoint Molecule TIGIT Could Help Tumor Growth in vivo by Suppressing the Function of NK and CD8+ T Cells.
TIGIT, an immune checkpoint molecule widely expressed on NK cells, activated T cells and Tregs, has been involved in delivering inhibitory signals through the interaction with PVR. The blockade of TIGIT/PVR interaction is a promising approach in cancer immunotherapy. Here, we unexpectedly discovered the expression of TIGIT in murine tumor cells. To elucidate the mechanism of such intrinsic expression, TIGIT knockout murine colorectal CT26 and MC38 cell lines were generated by using CRISPR/Cas9 system. Although TIGIT knockout showed no effects on proliferation and colony formation of tumor cells in vitro, the tumor growth in mice was considerably inhibited. TIGIT knockout led to the increase of IFN-γ secretion by NK and CD8+ T cells. Further, in BABL/c nude mice, CD8+ T cells depleting mice and NK cells depleting nude mice, the promotion of tumor growth was significantly diminished, suggesting that both NK cells and CD8+ T cells were involved in the tumor promoting process mediated by intrinsic TIGIT. In addition, blocking TIGIT/PVR interaction by the antibody or recombinant PVR protein could elicit anti-tumor effects by facilitating the tumor infiltration and restoring the function of CD8+ T cells, and the antibody-mediate TIGIT blockade could inhibit MC38 tumor growth through blocking TIGIT expressed on tumor cells. We therefore propose a novel TIGIT/PVR interaction mode that tumor intrinsic TIGIT delivers inhibitory signals to CD8+ T cells and NK cells by engaging with PVR.


Natural history of atypical and equivocal melanocytic lesions in children: an observational study of 19 cases.
Digital dermoscopy follow-up helps to identify patterns of change typical of common atypical nevi and early melanoma and improves the follow-up of patients with atypical nevi. We report the morphologic changes observed over time in 19 atypical or equivocal acquired melanocytic nevi that underwent dermoscopic follow-up. Two observers retrospectively examined digitalized dermoscopic images of 19 atypical melanocytic nevi from 15 children and young adults (median age 12 years, range 3-26 years). The images were assessed for global dermoscopic patterns at baseline and after a median 25-month (range 6-138 mos) follow-up. Ten (52.6%) nevi changed and nine (47.4%) retained a stable dermoscopic pattern. Of the 10 changing lesions, 2 of 4 homogeneous nevi evolved into a reticular pattern and 2 into a mixed pattern; 1 of 2 nevi with a mixed pattern evolved into a homogeneous nevus and 1 into a regressing nevus; 1 of 2 nevi with "other" patterns, such as negative pigment network and peppering throughout the lesion, evolved into a mixed nevus and 1 into a regressing nevus; 1 globular nevus evolved into a mixed pattern; and 1 starburst nevus evolved into a homogeneous nevus. The most striking results of our study were that atypical nevi can evolve into common nevi or they can regress, as documented by long-term dermoscopic follow-up. In children and young adults, dermoscopic follow-up of atypical nevi might be a valid alternative to surgical excision and enables us to achieve new insights into the natural history of these nevi.


The expression of microRNA miR-107 decreases early in Alzheimer's disease and may accelerate disease progression through regulation of beta-site amyloid precursor protein-cleaving enzyme 1.
MicroRNAs (miRNAs) are small regulatory RNAs that participate in posttranscriptional gene regulation in a sequence-specific manner. However, little is understood about the role(s) of miRNAs in Alzheimer's disease (AD). We used miRNA expression microarrays on RNA extracted from human brain tissue from the University of Kentucky Alzheimer's Disease Center Brain Bank with near-optimal clinicopathological correlation. Cases were separated into four groups: elderly nondemented with negligible AD-type pathology, nondemented with incipient AD pathology, mild cognitive impairment (MCI) with moderate AD pathology, and AD. Among the AD-related miRNA expression changes, miR-107 was exceptional because miR-107 levels decreased significantly even in patients with the earliest stages of pathology. In situ hybridization with cross-comparison to neuropathology demonstrated that particular cerebral cortical laminas involved by AD pathology exhibit diminished neuronal miR-107 expression. Computational analysis predicted that the 3'-untranslated region (UTR) of beta-site amyloid precursor protein-cleaving enzyme 1 (BACE1) mRNA is targeted multiply by miR-107. From the same RNA material analyzed on miRNA microarrays, mRNA expression profiling was performed using Affymetrix Exon Array microarrays on nondemented, MCI, and AD patients. BACE1 mRNA levels tended to increase as miR-107 levels decreased in the progression of AD. Cell culture reporter assays performed with a subset of the predicted miR-107 binding sites indicate the presence of at least one physiological miR-107 miRNA recognition sequence in the 3'-UTR of BACE1 mRNA. Together, the coordinated application of miRNA profiling, Affymetrix microarrays, new bioinformatics predictions, in situ hybridization, and biochemical validation indicate that miR-107 may be involved in accelerated disease progression through regulation of BACE1.


A functional analysis of disease-associated mutations in the androgen receptor gene.
Mutations in the androgen receptor (AR) are associated with a variety of diseases including androgen insensitivity syndrome and prostate cancer, but the way in which these mutations cause disease is poorly understood. We present a method for distinguishing likely disease-causing mutations from mutations that are merely associated with disease but have no causal role. Our method uses a measure of nucleotide conservation, and we find that conservation often correlates with severity of the clinical phenotype. Further, by only including mutations whose pathogenicity has been proven experimentally, this correlation is enhanced in the case of prostate cancer-associated mutations. Our method provides a means for assessing the significance of single nucleotide polymorphisms (SNPs) and cancer-associated mutations.


Acs in paediatrics.
Abdominal Compartment Syndrome (ACS) occurs relatively infrequent in a paediatric population when compared with adults. Overall mortality is still high. Also, the pathophysiologic mechanism that leads to ACS is different in children. In this review, we will present an overview on ACS in children admitted to a paediatric intensive care unit.


18F-FDG PET/CT Findings Following Repeated Intramuscular Injections of "Site Enhancement Oil" in the Upper Extremities.
We present the findings on F-FDG PET/CT in a 50-year-old man known to self-administer intramuscular injections with site enhancement oil in the upper extremities. PET images show diffuse pathological high FDG uptake in soft tissue of the upper arms and in scanned portions of the forearms. On the CT images, the muscles in the upper arm are swollen with a moth-eaten appearance and surrounding edema. Interspersed in the muscle tissue are several "cystic" lesions interpreted as oil deposits termed oleomas.


Linear kinematics at take-off in horses jumping the wall in an international Puissance competition.
Sagittal plane SVHS video recordings (50 Hz) were made of horses jumping the wall at an international Puissance competition. Video sequences were manually digitized and six kinematic variables at take-off were analyzed. Nine horses started the competition with the fence height at 1.80 m, and two horses attempted the fence in the fifth and final round with the fence height at 2.27 m. For successful performances, fence height was correlated with the following take-off variables: vertical velocity of the centre of mass (r = 0.45, p = 0.03); height of centre of mass (r = 0.44, p = 0.04); distance of centre of mass from fence (r = 0.46, p = 0.03); and distance from leading hind limb to centre of mass (r = -0.61, p < 0.01). These results indicated that body position at take-off is the most important aspect when jumping high fences. This is the first known study that has examined horses jumping over a Puissance wall. The results should help horse riders and trainers improve performance in Puissance jumping horses, and perhaps help in the early selection of horses with a talent for jumping high fences.


Objectively Measured Physical Activity and Polypharmacy Among Brazilian Community-Dwelling Older Adults.
Previous observations regarding association between physical activity (PA) and use of medicines among older adults are derived from self-reported PA. This study aimed to evaluate the association between objectively measured PA and polypharmacy among older adults with multimorbidity in Southern Brazil. This study included 875 noninstitutionalized older people, aged ≥60 years. Prescribed medicines used in the 15 days prior to the interview, socioeconomic data, and the presence of comorbidities were self-reported. Accelerometers were used to evaluate PA following the interview. Prevalence of polypharmacy (≥5 medicines) was 38.3% (95% confidence interval, 35.0-41.5); those belonging to the lowest tertile of PA used more medicines. The authors observed a significant inverse association for polypharmacy between men belonging to the second and third tertiles of PA for objectively measured overall PA and light PA compared with the most inactive tertile. For women, the association between PA and polypharmacy was significant for overall, light, and moderate to vigorous PA only in the third tertile. Overall, light and moderate to vigorous PA were inversely associated to polypharmacy and differed by gender. Promotion of PA in older adults may be an effective intervention to reduce the number of medicines used independent of the number of comorbidities.


Statin Use and Reduced Risk of Pneumonia in Melioidosis Patients: A Lung-Specific Statin Association.
HMG-CoA reductase inhibitor (statin) use is associated with a lower risk of incident pneumonia and, less robustly, with non-pulmonary infections. Whether statin use is associated with a lower risk of pneumonia compared to other clinical presentations of infection with the same pathogen is unknown. To assess whether pre-admission statin use is associated with a lower risk of pneumonia compared to non-pneumonia presentations among patients hospitalized with Burkholderia pseudomallei infection (melioidosis). We performed a secondary analysis of a prospective multicenter cohort study of patients hospitalized with culture-confirmed melioidosis. We used Poisson regression with robust standard errors to test for an association between statin use and pneumonia. We then performed several sensitivity analyses that addressed healthy user effect and indication bias. Of 1372 patients enrolled in the parent cohort, 1121 were analyzed. 980/1121 (87%) were statin nonusers and 141/1121 (13%) were statin users. 46/141 (33%) statin users presented with pneumonia compared with 432/980 (44%) of statin nonusers. Statin use was associated with a lower risk of pneumonia in unadjusted analysis (relative risk (RR) 0.74, 95% CI 0.58-0.95, p=0.02) and after adjustment for demographic variables, comorbidities, and environmental exposures (RR 0.73; 95% CI 0.57-0.94; p=0.02). Sensitivity analyses including active comparator analysis and inverse probability of treatment weighting were consistent with the primary analysis. In hospitalized melioidosis patients, pre-admission statin use was associated with a lower risk of pneumonia compared with other clinical presentations of melioidosis, suggesting a lung-specific protective effect of statins.


Pregnancy is associated with elevation of liver enzymes in HIV-positive women on antiretroviral therapy.
The objective of this study is to assess whether pregnancy is associated with an increased risk of liver enzyme elevation (LEE) and severe LEE in HIV-positive women on antiretroviral therapy (ART). Two observational studies: the UK Collaborative HIV Cohort (UK CHIC) study and the UK and Ireland National Study of HIV in Pregnancy and Childhood (NSHPC). Combined data from UK CHIC and NSHPC were used to identify factors associated with LEE (grade 1-4) and severe LEE (grade 3-4). Women starting ART in 2000-2012 were included irrespective of pregnancy status. Cox proportional hazards were used to assess fixed and time-dependent covariates including pregnancy status, CD4 cell count, drug regimen and hepatitis B virus/hepatitis C virus (HBV/HCV) coinfection. One-quarter (25.7%, 982/3815) of women were pregnant during follow-up, 14.2% (n = 541) when starting ART. The rate of LEE was 14.5/100 person-years in and 6.0/100 person-years outside of pregnancy. The rate of severe LEE was 3.9/100 person-years in and 0.6/100 person-years outside of pregnancy. The risk of LEE and severe LEE was increased during pregnancy [LEE: adjusted hazard ratio (aHR) 1.66 (1.31-2.09); severe LEE: aHR 3.57 (2.30-5.54)], including in secondary analyses excluding 541 women pregnant when starting ART. Other factors associated with LEE and severe LEE included lower CD4 cell count (<250 cells/μl), HBV/HCV coinfection and calendar year. Although few women developed severe LEE, this study provides further evidence that pregnancy is associated with an increased risk of LEE and severe LEE, reinforcing the need for regular monitoring of liver biomarkers during pregnancy.


Interaction of phenylthiolato-(2,2',2"-terpyridine)platinum(II) cation with DNA.
The interaction between a novel aromatic thiolato derivative from the family of DNA-intercalating platinum complexes, phenylthiolato-(2,2',2"-terpyridine)platinum(II)-[PhS(ter py)Pt+], and nucleic acids was studied by using viscosity, equilibrium-dialysis and kinetic measurements. Viscosity measurements with sonicated DNA provide direct evidence for intercalation, and show that at binding ratios below 0.2 molecules per base-pair PhS(terpy)Pt+ causes an increase in contour length of 0.2 nm per bound molecule. However, helix extension diminishes at greater extents of binding, indicating the existence of additional, non-intercalated, externally bound forms of the ligand. The ability of PhS(terpy)Pt+ to aggregate in neutral aqueous buffers at a range of ionic strengths and temperatures was assessed by using optical-absorption methods. Scatchard plots for binding to calf thymus DNA at ionic strength 0.01 (corrected for dimerization) are curvilinear, concave upward, providing further evidence for two modes of binding. The association constant decreases at higher ionic strengths, in accord with the expectations of polyelectrolyte theory, although the number of cations released per bound unipositive ligand molecule is substantially greater than 1. Stopped-flow kinetic measurements confirm the complexity of the binding reaction by revealing multiple bound forms of the ligand whose kinetic processes are both fast and closely coupled. Thermal denaturation of DNA radically alters the shapes of binding isotherms and either has little effect on, or enhances, the affinity of potential binding sites, depending on experimental conditions. Scatchard plots for binding to natural DNA species with differing nucleotide composition show that the ligand has a requirement for a single G X C base-pair at the highest-affinity intercalation sites.


Real-time fluorescence assays to monitor duplex unwinding and ATPase activities of helicases.
Many physiological functions of helicases are dependent on their ability to unwind nucleic acid duplexes in an ATP-dependent fashion. Determining the kinetic frameworks of these processes is crucial to understanding how these proteins function. We recently developed a fluorescence assay to monitor RNA duplex unwinding by DEAD-box helicases in real time. In this assay, two fluorescently modified short reporter oligonucleotides are annealed to an unmodified RNA loading strand of any length so that the fluorescent moieties of the two reporters find themselves in close proximity to each other and fluorescence is quenched. One reporter is modified with cyanine 3 (Cy3), whereas the other is modified with a spectrally paired black-hole quencher (BHQ). As the helicase unwinds the loading strand, the enzyme displaces the Cy3-modified reporter, which will bind to a capture or competitor DNA strand, permanently separating it from the BHQ-modified reporter. Complete separation of the Cy3-modified reporter strand is thus detected as an increase in total fluorescence. This assay is compatible with reagentless biosensors to monitor ATPase activity so that the coupling between ATP hydrolysis and duplex unwinding can be determined. With the protocol described, obtaining data and analyzing results of unwinding and ATPase assays takes ∼4 h.


[Micrometric in situ marking apparatus for an object on a microscope grid. Application of observation of serial sections].
A device for the precise localization (better than 0.5 micron) of an object on a grid has been developed and connected to the translation system of an electron microscope. Applied to biological thin sections, this device enables to easily find and to observe a selected microcrystal projection obtained by the grid sectioning technique. Moreover, low dose observations on ribbons of ultrathin sections can be made because the information is repeated in the successive sections of the ribbon: With the device the position of the selected object is determined at high dose in one section and low dose measurement is made on the following section.


Ca2+ influx through P2X1 receptors amplifies P2Y1 receptor-evoked Ca2+ signaling and ADP-evoked platelet aggregation.
Many cells express both P2X cation channels and P2Y G-protein-coupled receptors that are costimulated by nucleotides released during physiologic or pathophysiologic responses. For example, during hemostasis and thrombosis, ATP-gated P2X1 channels and ADP-stimulated P2Y1 and P2Y12 G-protein coupled receptors play important roles in platelet activation. It has previously been reported that P2X1 receptors amplify P2Y1-evoked Ca(2+) responses in platelets, but the underlying mechanism and influence on function is unknown. In human platelets, we show that maximally activated P2X1 receptors failed to stimulate significant aggregation but could amplify the aggregation response to a submaximal concentration of ADP. Costimulation of P2X1 and P2Y1 receptors generated a superadditive Ca(2+) increase in both human platelets and human embryonic kidney 293 (HEK293) cells via a mechanism dependent on Ca(2+) influx rather than Na(+) influx or membrane depolarization. The potentiation, due to an enhanced P2Y1 response, was observed if ADP was added up to 60 seconds after P2X1 activation. P2X1 receptors also enhanced Ca(2+) responses when costimulated with type 1 protease activated and M1 muscarinic acetylcholine receptors. The P2X1-dependent amplification of Gq-coupled [Ca(2+)]i increase was mimicked by ionomycin and was not affected by inhibition of protein kinase C, Rho-kinase, or extracellular signal-regulated protein kinase 1/2, which suggests that it results from potentiation of inositol 1,4,5-trisphosphate receptors and/or phospholipase C. We conclude that Ca(2+) influx through P2X1 receptors amplifies Ca(2+) signaling through P2Y1 and other Gq-coupled receptors. This represents a general form of co-incidence detection of ATP and coreleased agonists, such as ADP at sites of vascular injury or synaptic transmitters acting at metabotropic Gq-coupled receptors.


Spatiotemporal patterns of gamma frequency oscillations tetanically induced in the rat hippocampal slice.
1. We used transverse and longitudinal rat hippocampal slices to study the synchronization of gamma frequency (> 20 Hz) oscillations, across distances of up to 4.5 mm. gamma oscillations were evoked in the CA1 region by tetanic stimulation at one or two sites simultaneously, and were associated with population spikes. Tetanic stimuli that were strong enough to induce oscillations were associated with depolarization of both pyramidal cells and interneurones, largely produced by activation of metabotropic glutamate receptors. 2. Computer simulations of gamma oscillations were also performed in a model with pyramidal cells and interneurones, arranged in a chain of five cell groups. This model had suggested previously that interneurone networks alone could generate synchronous gamma oscillations locally, but that pyramidal cell firing, by inducing spike doublets in interneurones, was necessary for the occurrence of highly correlated oscillations with small phase lag (< 2.5 ms), in a distributed network possessing long axon conduction delays. 3. In both experiment and model, pyramidal cell spikes occurred in phase with local population spikes, as did the first spike of the interneurone doublet. 4. The conductance of the interneurone alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid (AMPA) receptor-mediated conductance was manipulated in the model, while the relation between oscillations at opposite ends of the chain was examined. When the conductance was large enough for doublet firing to be synaptically induced in interneurones, oscillation phase lags were < 2.25 ms across the chain. As predicted, experimental blockade of AMPA receptors resulted in increased phase lags between two sites oscillating simultaneously, compared with control conditions. 5. Both in model and in experiment, when stimuli to the two ends of the network were slightly different, cross-network synchronization occurred with a shorter phase lag at high frequencies than at lower frequencies. 6. These data suggest that, while interneurone networks alone can generate locally synchronized gamma oscillations, firing of pyramidal cells, and the synaptically induced doublet firing in interneurones, contribute to the stability and tight synchrony of the oscillations in distributed networks.


Smoking habits and obesity in young adults.
The aim of this work was to study the association between obesity and smoking habits in young adults. Specifically, we tested the hypothesis that obesity does not prevent young adults from smoking and conversely smoking does not protect against obesity. Trained nurses interviewed participants concerning demographic data and health behaviors such as smoking. At the time of the interview, weight and height were measured. Data were analyzed retrospectively. A representative sample of Israel Defense Force (IDF) personnel upon discharge from compulsory service, usually at the age of 20-21 years. Overall, 29 745 participants were included during the 13-year study (16,363 males and 13,382 females). Smoking rates were higher among obese participants than among overweight and non-obese participants (34.9%, 37.1%, 43.6% for non-obese, overweight and obese, respectively; P < 0.001). Mean number of cigarettes smoked per day were also higher among smokers that were obese and overweight compared to the non-obese (15.2 +/- 9.2, 15.6 +/- 10.7, 18.0 +/- 9.8, respectively; P < 0.001). Overweight and obesity were associated with the father's lower academic educational level. In logistic regression analysis, obesity, year of study and parental academic education were correlated independently with smoking (P < 0.001). The positive association between obesity and smoking suggests that obesity is not a deterrent to smoking and also that smoking does not help to prevent obesity.


[A multicenter randomized controlled study of the efficacy and safety of oseltamivir in the treatment of influenza in a high risk population].
To evaluate the efficacy and safety of oseltamivir in the treatment of influenza in a high risk population. A randomized, open, control trial was conducted from Nov. 2002 to Feb. 2003. Patients with chronic respiratory disease, such as chronic bronchitis, obstructive emphysema, bronchial asthma, bronchiectasis or chronic cardiac disease, and with symptoms of influenza were enrolled. They should satisfy the following criteria: Fever > or = 37.8 degrees C plus at least two of the following influenza symptoms: coryza/nasal congestion, sore throat, cough, myalgia/muscle aches and pain, fatigue, headache and chills/sweats. Within 48 h after the onset of the symptoms, the patients were randomly assigned to oseltamivir group (oseltamivir 75 mg, twice daily for 5 days) or control group (symptom relief medicine only). Fifty-six of the 108 recruited patients were identified as influenza-infected through laboratory test. They were defined as intent-to-treat infected population (ITTI) (27 oseltamivir, 29 control). The duration of influenza symptom was 64 h shorter (36.7%) and AUC score of the influenza symptom was decreased by 618 (43.1%) in the oseltamivir group as compared with those in the control group. The fever duration was 46.8 h (45.0%) less in the oseltamivir group than that in the control group. It took 6 d for the oseltamivir group and 11 days for the control group to recover to the basic health status. Secondary complications such as bronchitis, sinusitis and pneumonia occurred 11% (3/27) in the oseltamivir group and 45% (13/29) in the control group. The treatment expense for influenza and its complication was 587.4 RMB in the oseltamivir group and 786.5 RMB in the control group, which showed no significant difference (P = 0.246). It is suggested that oseltamivir is effective and well tolerated in patients with chronic respiratory or cardiac diseases. It can reduce the fever duration and severity of influenza symptom, and decrease the incidence of secondary complications and antibiotic use, while does not increase the total medical cost.


Glycemic Measures and Development and Resolution of Nonalcoholic Fatty Liver Disease in Nondiabetic Individuals.
Type 2 diabetes (T2D) is closely associated with nonalcoholic fatty liver disease (NAFLD); however, evidence regarding the link between blood glucose, especially below the threshold for T2D, and NAFLD is scarce. The objective of this work is to examine the associations of fasting glucose, oral glucose tolerance test (OGTT) 2-hour glucose, and hemoglobin A1c (HbA1c), and changes in these measures with development and resolution of NAFLD in nondiabetic individuals. This longitudinal cohort study comprised 4273 Chinese adults age 40 years or older and free of baseline T2D from 2010 to 2015. Blood sampling was performed during the OGTT test. NAFLD was ascertained by hepatic ultrasonography. Risk ratios (RRs) were calculated using modified Poisson regression models. During a mean 4.4 years of follow-up, NAFLD occurred in 573 (17.9%) of the 3209 participants without baseline NAFLD and resolved in 304 (28.6%) of the 1064 participants with baseline NAFLD. OGTT 2-h glucose was positively associated with NAFLD incidence (RR per 1-SD increase: 1.16, 95% CI: 1.08-1.25), whereas fasting (RR: 0.86, 95% CI: 0.78-0.94) and 2-hour glucose (RR: 0.85, 95% CI: 0.77-0.93) were inversely associated with resolution of NAFLD. Glycemic deterioration conferred increased risk of developing NAFLD and decreased likelihood of resolution of NAFLD than maintaining normal glycemic regulation (NGR). The strongest associations were observed for individuals who developed T2D. Meanwhile, baseline or incident NAFLD significantly increased the risk of deterioration in glucose metabolism. Increased glycemic levels within the nondiabetic range, as well as progression from NGR to T2D or prediabetes, were adversely associated with development and improvement of NAFLD.


Immune status and survival of opiate- and cocaine-treated mice infected with Friend virus.
In as much as the immunomodulatory effects of opiates and cocaine are known to modify spontaneous host defenses against infection, we investigated the effects of morphine, pentazocine and cocaine on the time course of Friend virus infection in mice. Repeated i.p. injections with increasing doses of morphine hydrochloride (10-100 mg/kg for 10 days before infection, a dose regimen which induced tolerance to the acute antinociceptive effects of the drug, followed by 30 mg/kg for 14 days postinfection) did not increase the mortality due to Friend virus infection. This regimen did not significantly affect the immune response of infected mice assessed in terms of delayed hypersensitivity (ear thickness) and the hemagglutination assay. In contrast, a single challenge with a large dose of morphine (up to 300 mg/kg), which is not lethal in noninfected mice, increased mortality markedly (up to 100%) in infected mice when administered at day 14 or 21 postinfection. Repeated i.p. injections with pentazocine (50 mg/kg b.i.d. for 5 days before infection, followed by 30 mg/kg for 14 days postinfection) had no influence on mortality or immune responses in infected mice; similar results were obtained with a single high-dose injection (up to 100 mg/kg). Lastly, repeated i.p. injections of cocaine, using the same experimental procedure as that for pentazocine, decreased immune responsiveness and slightly increased mortality, whereas a single injection was devoid of lethal effect. These findings suggest that chronic opioid treatment does not lower host resistance to viral infection but that the latter could increase the toxicity of a single high dose of morphine.(ABSTRACT TRUNCATED AT 250 WORDS)


Integration of signals from receptor tyrosine kinases and g protein-coupled receptors.
Activation of G protein-coupled receptors (GPCRs) leads to stimulation of classical G protein signaling pathways. In addition, GPCRs can activate the mitogen-activated protein kinases (MAPKs) such as the extracellular signal-regulated kinases, c-Jun NH(2)-terminal kinases (JNKs), and p38 MAPKs, and thereby influence cell proliferation, cell differentiation and mitogenesis. Cross talk between GPCRs and receptor tyrosine kinases (RTKs) is an incredibly complex process, and the exact signaling molecules involved are largely dependent on the cell type and the type of receptor that is activated. In this review we investigate recent advances that have been made in understanding the mechanisms of cross talk between GPCRs and RTKs, with a focus on GPCR-mediated activation of the Ras/MAPK pathway, GPCR-induced transactivation of RTKs, GPCR-mediated activation of JNK, and p38 MAPK, integration of signals by RhoGTPases, and activation of G protein signaling pathways by RTKs.


Effect of tree nuts on glycemic control in diabetes: a systematic review and meta-analysis of randomized controlled dietary trials.
Tree nut consumption has been associated with reduced diabetes risk, however, results from randomized trials on glycemic control have been inconsistent. To provide better evidence for diabetes guidelines development, we conducted a systematic review and meta-analysis of randomized controlled trials to assess the effects of tree nuts on markers of glycemic control in individuals with diabetes. MEDLINE, EMBASE, CINAHL, and Cochrane databases through 6 April 2014. Randomized controlled trials ≥3 weeks conducted in individuals with diabetes that compare the effect of diets emphasizing tree nuts to isocaloric diets without tree nuts on HbA1c, fasting glucose, fasting insulin, and HOMA-IR. Two independent reviewer's extracted relevant data and assessed study quality and risk of bias. Data were pooled by the generic inverse variance method and expressed as mean differences (MD) with 95% CI's. Heterogeneity was assessed (Cochran Q-statistic) and quantified (I2). Twelve trials (n = 450) were included. Diets emphasizing tree nuts at a median dose of 56 g/d significantly lowered HbA1c (MD = -0.07% [95% CI:-0.10, -0.03%]; P = 0.0003) and fasting glucose (MD = -0.15 mmol/L [95% CI: -0.27, -0.02 mmol/L]; P = 0.03) compared with control diets. No significant treatment effects were observed for fasting insulin and HOMA-IR, however the direction of effect favoured tree nuts. Majority of trials were of short duration and poor quality. Pooled analyses show that tree nuts improve glycemic control in individuals with type 2 diabetes, supporting their inclusion in a healthy diet. Owing to the uncertainties in our analyses there is a need for longer, higher quality trials with a focus on using nuts to displace high-glycemic index carbohydrates. ClinicalTrials.gov NCT01630980.


[Syndromes 10. Beckwith-Wiedemann syndrome].
Beckwith-Wiedemann syndrome is characterised by variable clinical combinations of exomphalos, macroglossia, gigantism, hypoglycemia and an increased incidence of tumors. This article briefly describes its clinical symptomatology. Recently, hemangiomatous lesions in tongues have been determined.


Sialic acid as the potential link between lipid metabolism and inflammation in the pathogenesis of atherosclerosis.
In the modern world, cardiovascular diseases have a special place among the most common causes of death. Naturally, this widespread problem cannot escape the attention of scientists and researchers. One of the main conditions preceding the development of fatal cardiovascular diseases is atherosclerosis. Despite extensive research into its pathogenesis and possible prevention and treatment strategies, many gaps remain in our understanding of this disease. For example, the concept of multiple low-density lipoprotein modifications was recently stated, in which desialylation is of special importance. Apart from this, sialic acids are known to be important contributors to processes such as endothelial dysfunction and inflammation, which in turn are major components of atherogenesis. In this review, we have collected information on sialic acid metabolism, analyzed various aspects of its implication in atherosclerosis at different stages, and provided an overview of the role of particular groups of enzymes responsible for sialic acid metabolism in the context of atherosclerosis.


The fitness cost of mismatch repair mutators in Saccharomyces cerevisiae: partitioning the mutational load.
Mutational load is the depression in a population's mean fitness that results from the continual influx of deleterious mutations. Here, we directly estimate the mutational load in a population of haploid Saccharomyces cerevisiae that are deficient for mismatch repair. We partition the load in haploids into two components. To estimate the load due to nonlethal mutations, we measure the competitive fitness of hundreds of randomly selected clones from both mismatch-repair-deficient and -proficient populations. Computation of the mean clone fitness for the mismatch-repair-deficient strain permits an estimation of the nonlethal load, and the histogram of fitness provides an interesting visualization of a loaded population. In a separate experiment, in order to estimate the load due to lethal mutations (i.e. the lethal mutation rate), we manipulate thousands of individual pairs of mother and daughter cells and track their fates. These two approaches yield point estimates for the two contributors to load, and the addition of these estimates is nearly equal to the separately measured short-term competitive fitness deficit for the mismatch-repair-deficient strain. This correspondence suggests that there is no need to invoke direct fitness effects to explain the fitness difference between mismatch-repair-deficient and -proficient strains. Assays in diploids are consistent with deleterious mutations in diploids tending towards recessivity. These results enhance our understanding of mutational load, a central population genetics concept, and we discuss their implications for the evolution of mutation rates.


Patient-matched instruments versus standard instrumentation in total knee arthroplasty: a prospective randomized study.
Optimal positioning of implants and restoration of neutral mechanical axis are two primary surgical goals in total knee arthroplasty (TKA). Despite modern instruments and improved surgical techniques, malalignment remains an important cause of early failure after TKA. The aim of this prospective randomized study was to compare the value of a new patient-matched instrument system (PMI) (Signature(TM); Biomet, Inc, Warsaw, Indiana) to that of standard TKA surgical instrumentation (STD) in terms of coronal mechanical alignment, time of operation and blood loss. A total of 38 patients waiting for primary TKA were enrolled and randomized into two groups (19 PMI and 19 STD). Magnetic resonance imaging was performed in all patients in the PMI group, and specific instruments for the femur and tibia were designed preoperatively. All patients were operated on using the standard medial parapatellar approach with no use of tourniquet. Mechanical axis, time for the operation, and blood loss were evaluated. Patients in both groups had comparable age, body mass index, preoperative mechanical axis, Knee Society Score, and level of hemoglobin. Postoperative results showed that the PMI group fell significantly closer to neutral mechanical axis (STD: 2.7 ± 1.7, PMI: 1.7 ± 0.9; P = 0.013) with no outliers and a reduced time for the operation. There was no difference in the evaluation blood loss. The use of PMI can contribute in achieving better mechanical axis with reduction in outliers and decreased operation time. Due to small differences between PMI and standard instruments, additional research are needed to confirm these preliminary results, and to discover potential benefits and functional improvements in the long-term outcome.


Psychophysics of reading--X. Effects of age-related changes in vision.
This study examined the influence of age-related changes in vision on reading performance. Maximum reading speed was measured in groups of young (n = 16, mean age 21.6 years) and old (n = 14, mean age 68.3 years) subjects, all with acuities of 20/32 or better. A psychophysical procedure was used for measuring reading speed that has proven reliable and sensitive to visual factors in previous research. Data were collected for character sizes ranging from .15 degrees to 12 degrees. Research revealed that old subjects who were free of eye disease read as fast as the young subjects for character sizes ranging from .3 degrees to 1.0 degrees. This is the range in which reading speed is maximum for young subjects. Research also revealed that old subjects showed a deficit when reading text composed of very small or very large characters. Their speeds dropped to about 70% of the young adult speeds. These deficits may be due to age-related losses in visual contrast sensitivity.


Titin-based regulations of diastolic and systolic functions of mammalian cardiac muscle.
Titin is the largest protein in mammals; it forms an elastic filament along the myofibril of cardiac and skeletal muscles. Novel studies employing the recently available varied technologies have revealed the molecular mechanisms by which titin generates passive force in the sarcomere in response to external stretch. Changes in titin stiffness occur during heart disease via a shift in the expression ratio of the two main titin isoforms, called N2B (stiff type) and N2BA (compliant type) titins. Protein kinase (PK)A, PKG and PKC phosphorylate the cardiac specific I-band titin segment, resulting in an acute decrease (by PKA and PKG) or increase (by PKC) in passive force. It has also been discovered that titin performs roles that go beyond passive force generation, by enhancing or terminating active force production, thereby adjusting the Frank-Starling mechanism of the heart. Therefore, titin is a self-adjustable and multi-functional spring that is indispensable for proper heart functions. Here, we discuss how titin regulates the passive and active properties of cardiac muscle in normal physiological conditions as well as in chronic heart disease.


The influence of fixed orthodontic appliances on masticatory and swallowing threshold performances.
To test the hypothesis that treatment with orthodontic appliances disturbs masticatory and swallowing performances. Twenty-seven subjects with malocclusions requiring orthodontic treatment were included in this prospective study. The masticatory and swallowing performances were evaluated at five different times: before bracket placement (T0), immediately after archwire placement (T1), 48 h after archwire placement (T2), 30 days after archwire placement (T3) and 3 months after the initial appointment (T4). Masticatory performance was determined by the median particle sizes for the Optocal test food after 15 chewing strokes, and the swallowing thresholds were registered for both the test food and a natural food (peanuts). Pain during mastication was evaluated using a 100-mm visual analogue scale. Masticatory performance was significantly reduced at T2, at which time patients reported the highest pain values. The time spent to the first swallow was increased at T2 for the natural food but not for the test food. The values for pain, masticatory and swallowing performances at T3 and T4 were similar to those at T0. Orthodontic patient masticatory function is only reduced during the period of higher pain experience, which could also disrupt the deglutition of harder foods. However, neither mastication nor deglutition processes were disturbed by orthodontic appliances in long-term treatment.


Incidence of symptomatic venous thromboembolism in oncological oral and maxillofacial operations: retrospective analysis.
We retrospectively analysed the incidence of symptomatic venous thromboembolism (VTE) and associated risk factors in operations under general anaesthesia for cancer of the oral cavity. To identify symptoms related to deep venous thrombosis (DVT) and pulmonary embolism (PE), together with associated risk factors, we reviewed medical records of patients operated on in the department of oral and maxillofacial surgery at the Queen Elizabeth Hospital, Birmingham, United Kingdom, between June 2007 and October 2012. All patients were categorised according to their level of risk of VTE. The incidence of VTE was calculated with univariate associations and odds ratios with related 95% confidence intervals, where possible. In total, 233 patients were included, comprising 244 operations (mean (SD) age at operation 60.9 (13) years). Almost all patients (97%) were classified as having the highest risk of VTE. Swelling of an extremity, expectoration of blood, and tightness of the chest were the most common symptoms for suspected cases. An incidence of 0.41% was found for symptomatic VTE; one man developed a PE 2 days after operation. Associations between the analysed factors and symptomatic VTE were not significant. The development of the complication in oncological oral and maxillofacial operations seems to be rare, even in patients with a high risk. We cannot recommend the use of routine thromboprophylaxis, but it could be advocated in patients with obvious serious risk factors.


Cerebral metastatic choriocarcinoma: intensive therapy and prognosis.
Choriocarcinoma is known to be sensitive to chemotherapy. Remission rates of 70% are reported for patients with metastatic disease. The Southeastern Regional Trophoblastic Disease Center of Duke University has reviewed its experience with the treatment of cerebral metastases from choriocarcinoma. Fourteen patients were identified as having cerebral metastases from a group of more than 500 patients with gestational trophoblastic disease (GTD) other than primary hydatidiform mole. The remission rate of 50% (7/14) was achieved by vigorous, multiagent chemotherapy and combined cerebral irradiation therapy. This series of patients is reviewed with regard to diagnosis, details of multiagent chemotherapy with cerebral irradiation, complications, and survival. The key factors for successful outcome seem to be early diagnosis and vigorous therapy.


Design, synthesis and efficacy of novel G protein-coupled receptor kinase 2 inhibitors.
G protein-coupled receptor kinase 2 (GRK2) is a relevant signaling node of the cellular transduction network, playing major roles in the physiology of various organs/tissues including the heart and blood vessels. Emerging evidence suggests that GRK2 is up regulated in pathological situations such as heart failure, hypertrophy and hypertension, and its inhibition offers a potential therapeutic solution to these diseases. We explored the GRK2 inhibitory activity of a library of cyclic peptides derived from the HJ loop of G protein-coupled receptor kinases 2 (GRK2). The design of these cyclic compounds was based on the conformation of the HJ loop within the X-ray structure of GRK2. One of these compounds, the cyclic peptide 7, inhibited potently and selectively the GRK2 activity, being more active than its linear precursor. In a cellular system, this peptide confirms the beneficial signaling properties of a potent GRK2 inhibitor. Preferred conformations of the most potent analog were investigated by NMR spectroscopy.


The growing recognition of immunotherapy-responsive seizure disorders with autoantibodies to specific neuronal proteins.
The concept of epilepsy and seizure disorders caused by autoantibodies to specific neuronal membrane proteins has developed significantly during the past few years. Antibodies to cell-surface membrane proteins such as voltage-gated potassium channels or N-methyl-D-aspartate receptors, or to glutamic acid decarboxylase, are found in patients with different forms of limbic encephalitis, and in a few patients with epilepsy as their main or only condition. Many of these patients do not show a good response to conventional antiepileptic drugs, but respond to immunotherapies. By contrast, studies of other antibodies in idiopathic forms of epilepsy, or epilepsy associated with systemic lupus erythematosus or coeliac disease, have not in general disclosed consistent, clinically helpful results. There are a growing number of specific antibodies associated with new onset epilepsy. These patients are likely to have an immune-mediated disorder that may benefit from immunotherapies. In autoimmune diseases such as systemic lupus erythematosus or coeliac disease, antibodies to specific membrane targets may also prove to be important in the future.


Labeling quinone-binding sites in photosynthetic reaction centers: A 38-kilodalton protein associated with the acceptor side of photosystem II.
2-Acetoxymethyl-1,4-naphthoquinone (2-AcOMeNQ) binds with rapid kinetics and high affinity to the primary quinone Q(A) site of reaction centers from Rhodopseudomonas capsulata. Binding of 2-AcOMeNQ fully restores electron-transfer activity with kinetics that is similar, but not identical, to that seen with ubiquinone-50. When bound at the Q(A) site, 2-AcOMeNQ preferentially labels the L subunit. This preference suggests that 2-AcOMeNQ labels primarily the region of a quinone-binding site that is close to the first isoprenoid unit of the side chain, which is expected from the location and structure of the reaction region of the molecule. In photosystem II particles from Synechococcus sp., 2-AcOMeNQ primarily labels two polypeptides with apparent molecular masses of 38 and 19 kDa. Labeling of only the 38-kDa polypeptide is sufficiently sensitive to 3-(3,4-dichlorophenyl)-1,1-dimethylurea (DCMU) to conclude that it is involved in binding quinones on the acceptor side of photosystem II. Although we have not yet identified the 38-kDa protein, its properties suggest that it is the D2 protein. From the DCMU-sensitive labeling and from homologies to functionally important regions of the bacterial reaction-center subunits, we propose that the 38-kDa protein is intimately involved in binding the cofactors that mediate primary photochemistry.


Transmucosal passage of bacteria across rat intestinal epithelium in the Ussing chamber: effect of nutritional factors and bacterial virulence.
Transmucosal passage of bacteria across the intestine, the essential and prerequisite step for bacterial translocation, cannot be effectively studied using in vivo models of translocation. We have adapted the Ussing chamber into a fresh, sterile organ culture system that can facilitate the study of bacterial-epithelial interactions. Intestinal membranes were mounted in the Ussing chamber and perfused with a solution rich in putative mucosal micronutrients. The transmembrane potential difference was constantly monitored as a marker of intestinal integrity. Transmucosal passage of various bacteria across the normal intestinal epithelium was quantitated, and the mucosal membrane was examined by light and transmission electron microscopy. The addition of potassium cyanide to the mucosal perfusate resulted in an irreversible loss of potential difference. Oxygen deprivation also led to a precipitous drop in potential difference, but it was reversible with prompt reoxygenation. In contrast, intestinal membranes perfused with a solution consisting of Dulbecco's modified Eagle's medium + 20 mM glutamine maintained their potential difference for a sustained period (>180 min). Both the viability and structural integrity of the ileal intestinal membrane were maintained in culture ex vivo using this perfusate. Qualitative differences were observed in the mechanism of transmucosal passage of mild to moderately virulent bacteria such as Escherichia coli C-25 and Proteus mirabilis M-13, which pass through the intestinal epithelium without causing overt damage to the mucosa, and more virulent organisms such as Salmonella typhimurium, which cause extensive mucosal damage by light and transmission electron microscopy. The Ussing system should provide a useful model of intact organ culture for the study of the mechanisms of bacterial translocation and the pathogenesis of enteric infections.


Chemotherapy in pediatric brain tumor and the challenge of the blood-brain barrier.
Pediatric brain tumors (PBT) stand as the leading cause of cancer-related deaths in children. Chemoradiation protocols have improved survival rates, even for non-resectable tumors. Nonetheless, radiation therapy carries the risk of numerous adverse effects that can have long-lasting, detrimental effects on the quality of life for survivors. The pursuit of chemotherapeutics that could obviate the need for radiotherapy remains ongoing. Several anti-tumor agents, including sunitinib, valproic acid, carboplatin, and panobinostat, have shown effectiveness in various malignancies but have not proven effective in treating PBT. The presence of the blood-brain barrier (BBB) plays a pivotal role in maintaining suboptimal concentrations of anti-cancer drugs in the central nervous system (CNS). Ongoing research aims to modulate the integrity of the BBB to attain clinically effective drug concentrations in the CNS. However, current findings on the interaction of exogenous chemical agents with the BBB remain limited and do not provide a comprehensive explanation for the ineffectiveness of established anti-cancer drugs in PBT. We conducted our search for chemotherapeutic agents associated with the blood-brain barrier (BBB) using the following keywords: Chemotherapy in Cancer, Chemotherapy in Brain Cancer, Chemotherapy in PBT, BBB Inhibition of Drugs into CNS, Suboptimal Concentration of CNS Drugs, PBT Drugs and BBB, and Potential PBT Drugs. We reviewed each relevant article before compiling the information in our manuscript. For the generation of figures, we utilized BioRender software. We focused our article search on chemical agents for PBT and subsequently investigated the role of the BBB in this context. Our search criteria included clinical trials, both randomized and non-randomized studies, preclinical research, review articles, and research papers. Our research suggests that, despite the availability of potent chemotherapeutic agents for several types of cancer, the effectiveness of these chemical agents in treating PBT has not been comprehensively explored. Additionally, there is a scarcity of studies examining the role of the BBB in the suboptimal outcomes of PBT treatment, despite the effectiveness of these drugs for other types of tumors.


Dopaminergic terminals in the rat cortex.
The destruction of ascending noradreniergic pathways by bilateral microinjections of 6-hydroxydopamnine made laterally to the pedunculus cerebellaris superior completely abolished the in vitro synthesis of [(3)H]norepinephrine from L-[(3)H]tyrosine in slices and in synaptosomes of the rat cortex. However, normal [(3)H]dopamine synthesis could still be observed in both cortical preparations from animals with lesions. These results provide the first biochemical support for the existence of dopaminergic terminals independent of noradrenergic terminals in the rat cortex.


NADPH oxidase deficiency underlies dysfunction of aged CD8+ Tregs.
Immune aging results in progressive loss of both protective immunity and T cell-mediated suppression, thereby conferring susceptibility to a combination of immunodeficiency and chronic inflammatory disease. Here, we determined that older individuals fail to generate immunosuppressive CD8+CCR7+ Tregs, a defect that is even more pronounced in the age-related vasculitic syndrome giant cell arteritis. In young, healthy individuals, CD8+CCR7+ Tregs are localized in T cell zones of secondary lymphoid organs, suppress activation and expansion of CD4 T cells by inhibiting the phosphorylation of membrane-proximal signaling molecules, and effectively inhibit proliferative expansion of CD4 T cells in vitro and in vivo. We identified deficiency of NADPH oxidase 2 (NOX2) as the molecular underpinning of CD8 Treg failure in the older individuals and in patients with giant cell arteritis. CD8 Tregs suppress by releasing exosomes that carry preassembled NOX2 membrane clusters and are taken up by CD4 T cells. Overexpression of NOX2 in aged CD8 Tregs promptly restored suppressive function. Together, our data support NOX2 as a critical component of the suppressive machinery of CD8 Tregs and suggest that repairing NOX2 deficiency in these cells may protect older individuals from tissue-destructive inflammatory disease, such as large-vessel vasculitis.


[Cell proliferation in mouse tissues after thymectomy and the administration of T-activin].
The epithelium of mouse cornea and lymph nodes was examined for DNA-synthetic and mitotic activity at different times after thymectomy and administration of T-activin, an active factor of the thymus. Thymectomy entails retardation of the rate of corneal epithelium regeneration, diminution in both tissues under study of the amplitude of oscillations in cell proliferation throughout the day. Administration to the animals of the immunoactive thymic factor T-activin makes the circadian rhythm of cell proliferation return to normal. It is assumed that T-activin raises the capacity of lymphocytes to interact with epithelial cells, which manifests itself in the enhancement of their mitotic activity.


The sensitivity of Porphyromonas gingivalis and Fusobacterium nucleatum to different (pseudo)halide-peroxidase combinations compared with mutans streptococci.
Previous studies have shown that the peroxidase system with iodide is particularly effective against Actinobacillus actinomycetemcomitans. In the present study, the effects of iodide, chloride and thiocyanate in combinations with lactoperoxidase (LP) and myeloperoxidase (MP) on the viability of Porphyromonas gingivalis, Fusobacterium nucleatum, Streptococcus mutans and S. rattus were analysed. Bacteria were incubated in buffer solution containing peroxidase, substrate(s) and H2O2 (all in oral physiological concentrations), and plated after 0, 0.5 and 1 h. The oxidation product of iodide was the most bactericidal against all the bacteria tested. The effect was significantly weaker on mutans streptococci. Physiological concentrations of thiocyanate abolished the effects of LP-H2O2-iodide and MP-H2O2-iodide/chloride combinations. Thiocyanate-peroxidase systems have already been used in oral hygiene products. The incorporation of iodide into these products could make them much more potent against periodontal pathogens, and also help to prevent transmission of these pathogens from person to person via saliva.


Methods to assess tissue-specific distribution and metabolism of drugs.
Most drugs exert their effects not within the plasma compartment, but in defined target tissues into which drugs have to distribute from the central compartment. Unfortunately, a complete and lasting equilibration between blood and tissue cannot always be taken for granted. Drug distribution processes may be characterized by a high intertissue- and intersubject variability and target site drug levels may substantially differ from corresponding plasma levels. Suboptimal target site concentrations may have important clinical implications, as it is a potential explanation for therapeutic failures. Therefore, determination of drug tissue penetration plays an important role in clinical drug development. In recent years, the assessment of tissue concentrations after administration of very low, sub-pharmacological drug amounts (microdosing) has attracted major interest in early clinical drug development, which calls for the availability of highly sensitive analytical methods. The present article will review the most important techniques that are currently available for studying drug disposition in humans. These can be classified as semi-invasive (microdialysis, MD) and non-invasive (positron emission tomography, PET, and magnetic resonance spectroscopy, MRS). We will discuss individual strengths and shortcomings of each method and provide some recent examples with particular focus on antiinfective and anticancer drugs. Whereas MD and MRS also lend themselves to the assessment of tissue-specific drug metabolism, PET usually does not provide metabolic information. For some drugs, such as the anticancer agents 5-fluorouracil and capecitabine, measurement of drug metabolites is particularly important as these represent the therapeutically active species.


Identification of a novel coronavirus in patients with severe acute respiratory syndrome.
The severe acute respiratory syndrome (SARS) has recently been identified as a new clinical entity. SARS is thought to be caused by an unknown infectious agent. Clinical specimens from patients with SARS were searched for unknown viruses with the use of cell cultures and molecular techniques. A novel coronavirus was identified in patients with SARS. The virus was isolated in cell culture, and a sequence 300 nucleotides in length was obtained by a polymerase-chain-reaction (PCR)-based random-amplification procedure. Genetic characterization indicated that the virus is only distantly related to known coronaviruses (identical in 50 to 60 percent of the nucleotide sequence). On the basis of the obtained sequence, conventional and real-time PCR assays for specific and sensitive detection of the novel virus were established. Virus was detected in a variety of clinical specimens from patients with SARS but not in controls. High concentrations of viral RNA of up to 100 million molecules per milliliter were found in sputum. Viral RNA was also detected at extremely low concentrations in plasma during the acute phase and in feces during the late convalescent phase. Infected patients showed seroconversion on the Vero cells in which the virus was isolated. The novel coronavirus might have a role in causing SARS.


Use of the chicken heart in a Langendorff preparation.
Hearts isolated from healthy chickens were perfused with oxygenated Krebs-Henseleit buffer. Balloons were inserted into the left ventricle, filled with saline and attached to a pressure transducer for measuring left ventricular pressures. A bipolar electrogram was obtained between the atria and ventricles. Hearts so perfused manifested mechanical alternans for approximately 15 minutes, after which they sustained a regular sinus rhythm without mechanical alternans for at least 60 minutes. By infusing 0.1 ml increments of saline into the balloon located within the ventricle, Frank-Starling curves could be generated. This preparation has potential for use in pharmacologic evaluations.


Is single port incisionless-intracorporeal conventional equipment-endoscopic surgery feasible in patients with retrocecal acute appendicitis?
Since laparoscopic appendectomy was first described, various modifications, such as single port incisionless-intracorporeal conventional equipment-endoscopic surgery (SPICES), have been described for reducing pain and improving cosmetic results. In the retrocecal and retrocolic positions, attachments to the lateral peritoneum and cecum may lead to difficulties during SPICES, which is performed with only one port. Here, we present the effects of variations in the position of the vermiform appendix in treating acute appendicitis with SPICES. We retrospectively reviewed 52 children who underwent SPICES for acute appendicitis between March 2010 and November 2011 in our institution. One group (group A) consisted of 30 patients (mean age, 10.5 ± 2.5 years) with retrocecal appendix, while the other group (group B) included 22 patients (mean age, 10.9 ± 2.3 years) with the appendix lying free in the peritoneal cavity. There were no significant differences between groups in terms of patient age, gender, success rate of SPICES, mean operating time, mean follow-up period, overall complication rates or mean postoperative hospitalization period. These results suggest that SPICES is a safe and feasible approach even in patients with retrocecal acute appendicitis.


Acute lymphoblastic leukemia in Weaver syndrome.
Weaver syndrome comprises pre- and postnatal overgrowth, accelerated osseous maturation, characteristic craniofacial appearance and developmental delay; it is a generally sporadic disorder, although autosomal dominant inheritance has been reported. Some of the manifestations characterize both the Weaver and Sotos syndrome, and distinction between the two is mainly by clinical examination and molecular testing. Most of the patients with Sotos syndrome have NSD1 gene deletions or mutations; however, the molecular basis of most of the Weaver syndrome patients is unknown. Patients with overgrowth syndromes have an increased frequency of tumors; the risk in Sotos syndrome patients has been estimated to be about 2-3%, with leukemia and lymphoma accounting for 44% of the malignancies. We report on a 4(1/2)-year-old girl with typical Weaver syndrome who developed acute lymphoblastic leukemia, an association not previously reported, and review the reported cases of Weaver syndrome patients who developed malignancies. Malignancy in Weaver syndrome has been reported previously in six patients. While searching the literature for all reported cases with Weaver syndrome and counting the cases with malignancy, we found that the frequency of tumors or hematologic malignancy was 10.9%. This is likely to be an overestimate, biased by failure to report cases without tumors and by over-reporting cases with this rare association. While the presence of acute lymphoblastic leukemia in our patient might be incidental, we cannot exclude a possible causative association between Weaver syndrome and hematologic malignancy.


Screening for transient ischemic attacks in hemodialysis patients.
Rapid recognition and management of transient ischemic attacks (TIAs) reduce incident strokes in the general population, but similar data are lacking in dialysis patients, who form a high-risk group for this pathology. We systematically screened hemodialysis patients for TIA to estimate its incidence and determine whether there was significant scope to reduce subsequent strokes by risk modification. Patients established on hemodialysis at a satellite dialysis unit at our center were screened prospectively using weekly symptom questionnaires over a 12-month period. Following clinical review, patients who screened positive were urgently referred to a TIA clinic, and all stroke and TIA and stroke events were recorded. A total of 304 patients were screened over 2,594 total patient months of follow-up (1st November 2009 to 1st November 2010). Six strokes occurred, of which 5 were ischemic (a rate of 23.1/1,000 patient-years). No patients screened positive for a TIA, despite predicted rates of 4.2/1,000 patient-years (95% confidence interval, 1.4-9.7/1,000 patient-years). One ischemic stroke was preceded by symptoms compatible with a TIA, although this was ascertained in retrospect and not during screening. Based on the first study of its kind to date, systematic screening for TIA has a low yield and cannot be relied on alone to identify patients at higher risk of cerebrovascular events. The confounding presence of symptoms attributable to uremia, neuropathy, hypotension and dysglycemia could reduce the sensitivity of established tests, with significant implications for the detection and treatment of TIA in dialysis.


Evolution of conformational order during self-assembly of n-alkanethiols on Hg droplets: an infrared spectromicroscopy study.
This Letter describes Fourier-transform infrared spectroscopy evidence for the evolution of conformational order and coverage during the formation of n-alkanethiol monolayers on microdroplets of mercury from the solution phase. At the highest coverages obtained by self-assembly, the monolayer is characterized by predominantly all-trans conformational order. For partial monolayers obtained at arbitrarily quenched incubation periods, we find a continuous evolution of the chain conformational order with monolayer coverage. Analyzing these results in light of previously reported models from X-ray scattering reveals a complex self-assembly process in which the density-dependent evolution of the chain conformational order is coupled with that of molecular orientation and density.


Genomics-Based Identification of Microorganisms in Human Ocular Body Fluid.
Advances in genomics have the potential to revolutionize clinical diagnostics. Here, we examine the microbiome of vitreous (intraocular body fluid) from patients who developed endophthalmitis following cataract surgery or intravitreal injection. Endophthalmitis is an inflammation of the intraocular cavity and can lead to a permanent loss of vision. As controls, we included vitreous from endophthalmitis-negative patients, balanced salt solution used during vitrectomy and DNA extraction blanks. We compared two DNA isolation procedures and found that an ultraclean production of reagents appeared to reduce background DNA in these low microbial biomass samples. We created a curated microbial genome database (>5700 genomes) and designed a metagenomics workflow with filtering steps to reduce DNA sequences originating from: (i) human hosts, (ii) ambiguousness/contaminants in public microbial reference genomes and (iii) the environment. Our metagenomic read classification revealed in nearly all cases the same microorganism that was determined in cultivation- and mass spectrometry-based analyses. For some patients, we identified the sequence type of the microorganism and antibiotic resistance genes through analyses of whole genome sequence (WGS) assemblies of isolates and metagenomic assemblies. Together, we conclude that genomics-based analyses of human ocular body fluid specimens can provide actionable information relevant to infectious disease management.


Spinal muscular atrophy: Broad disease spectrum and sex-specific phenotypes.
Spinal muscular atrophy (SMA) is one of the major genetic disorders associated with infant mortality. More than 90% of cases of SMA result from deletions of or mutations in the Survival Motor Neuron 1 (SMN1) gene. SMN2, a nearly identical copy of SMN1, does not compensate for the loss of SMN1 due to predominant skipping of exon 7. The spectrum of SMA is broad, ranging from prenatal death to infant mortality to survival into adulthood. All tissues, including brain, spinal cord, bone, skeletal muscle, heart, lung, liver, pancreas, gastrointestinal tract, kidney, spleen, ovary and testis, are directly and/or indirectly affected in SMA. Accumulating evidence on impaired mitochondrial biogenesis and defects in X chromosome-linked modifying factors, coupled with the sexual dimorphic nature of many tissues, point to sex-specific vulnerabilities in SMA. Here we review the role of sex in the pathogenesis of SMA.


Toxicity of car exhausts and opportunity for suicide: comparison between Britain and the United States.
The rate of car exhaust suicides in the United States has declined following the introduction of emission controls in the mid-1960s, though not as much as the decline in CO emitted by cars. In Britain, where emission controls have not been introduced, the rate of these suicides, initially much lower than in the United States, has greatly increased since the beginning of the 1970s and is now about double that of the United States. This rise cannot be explained simply on the basis of an increase in the opportunities for suicide as represented by an increase in the number of cars but may be due to increased knowledge of the method. While these results are interpreted as generally supporting the potential for opportunity-reducing preventive measures, they also demonstrate that much more research is needed into the complex nature of the opportunity structure for suicide.


Genetic control of the cell-division cycle in yeast. I. Detection of mutants.
Time-lapse photomicroscopy has been utilized to detect temperature-sensitive yeast mutants that are defective in gene functions needed at specific stages of the cell-division cycle. This technique provides two types of information about a mutant: the time at which the defective gene function is normally performed, defined as the execution point, and the stage at which cells collect when the function is not performed, defined as the termination point. Mutants carrying lesions in three genes that control the cell-division cycle are described. All three genes, cdc-1, cdc-2, and cdc-3, execute early in the cell cycle at about the time of bud initiation, but differ in their termination points. Cells carrying the cdc-1 mutation terminate at the execution point, most cells ending up with a tiny bud that does not develop further. Cells carrying the cdc-2 mutation terminate at mitosis. Cells carrying the cdc-3 mutation are defective in cell separation but show no definite termination point since other processes of the cell cycle, such as bud initiation and nuclear division, continue despite the block in cell separation.


Autoimmune diseases and infections as risk factors for schizophrenia.
Immunological hypotheses have become increasingly prominent when studying the etiology of schizophrenia. Autoimmune diseases, and especially the number of infections requiring hospitalization, have been identified as significant risk factors for schizophrenia in a dose-response relationship, which seem compatible with an immunological hypothesis for subgroups of patients with schizophrenia. Inflammation and infections may affect the brain through many different pathways that are not necessarily mutually exclusive and can possibly increase the risk of schizophrenia in vulnerable individuals. However, the findings could also be an epiphenomenon and not causal, due to, for instance, common genetic vulnerability, which could be supported by the observations of an increased prevalence of autoimmune diseases and infections in parents of patients with schizophrenia. Nevertheless, autoimmune diseases and infections should be considered in the treatment of individuals with schizophrenia symptoms, and further research is needed of the immune system's possible contributing pathogenic factors in the etiology of schizophrenia.


Single-molecule detection of dihydroazulene photo-thermal reaction using break junction technique.
Charge transport by tunnelling is one of the most ubiquitous elementary processes in nature. Small structural changes in a molecular junction can lead to significant difference in the single-molecule electronic properties, offering a tremendous opportunity to examine a reaction on the single-molecule scale by monitoring the conductance changes. Here, we explore the potential of the single-molecule break junction technique in the detection of photo-thermal reaction processes of a photochromic dihydroazulene/vinylheptafulvene system. Statistical analysis of the break junction experiments provides a quantitative approach for probing the reaction kinetics and reversibility, including the occurrence of isomerization during the reaction. The product ratios observed when switching the system in the junction does not follow those observed in solution studies (both experiment and theory), suggesting that the junction environment was perturbing the process significantly. This study opens the possibility of using nano-structured environments like molecular junctions to tailor product ratios in chemical reactions.


A chromatin landmark and transcription initiation at most promoters in human cells.
We describe the results of a genome-wide analysis of human cells that suggests that most protein-coding genes, including most genes thought to be transcriptionally inactive, experience transcription initiation. We found that nucleosomes with H3K4me3 and H3K9,14Ac modifications, together with RNA polymerase II, occupy the promoters of most protein-coding genes in human embryonic stem cells. Only a subset of these genes produce detectable full-length transcripts and are occupied by nucleosomes with H3K36me3 modifications, a hallmark of elongation. The other genes experience transcription initiation but show no evidence of elongation, suggesting that they are predominantly regulated at postinitiation steps. Genes encoding most developmental regulators fall into this group. Our results also identify a class of genes that are excluded from experiencing transcription initiation, at which mechanisms that prevent initiation must predominate. These observations extend to differentiated cells, suggesting that transcription initiation at most genes is a general phenomenon in human cells.


A "Universal" Spectroscopic Map for the OH Stretching Mode in Alcohols.
Empirical maps are presented for the OH stretching vibrations in neat alcohols in which the relevant spectroscopic quantities are expressed in terms of the electric field exerted on the hydrogen atom by the surrounding liquid. It is found, by examination of the four lowest linear alcohols, methanol, ethanol, n-propanol, and n-butanol, that a single map can be used for alcohols with different alkyl groups. This "universal" map is in very good agreement with maps optimized for the individual alcohols but differs from those previously developed for water. This suggests that one map can be used for all alcohols, perhaps even those not examined in the present study. The universal map gives IR lineshapes in good agreement with measured spectra for isotopically dilute methanol and ethanol, while the two-dimensional IR photon echo spectra give results that differ from experiments. The role of non-Condon effects, reorientation dynamics, hydrogen bonding, and spectral diffusion is discussed.


Outpatient treatment of thyroid cancer using high doses of iodine 131.
To develop a protocol for outpatient high-dose iodine 131 therapy to treat thyroid cancer, to allow therapy to be performed at the patient's convenience and save considerable hospital costs without causing undue radiation exposure to members of the public. The radiation safety hazard--both external (from gamma rays emitted from the patient) and internal (from radioactive material in the patient's excretions)--can be calculated according to guides issued by the US Nuclear Regulatory Commission. The protocol aimed to produce an "occupancy factor" of 0.125 or less for the "critical person" (person caring most for the patient) for the entire 72-hour period after administration of 131I. This is the equivalent of 3 hours per day at a distance of 1 m from the patient. Each patient's suitability for outpatient therapy is determined on the basis of the patient's home environment, ability to understand the risks involved and likelihood of compliance, by the referring physician's opinion, through a self-report questionnaire and through a patient interview with the radiation safety officer and the nuclear medicine physician. This protocol has been approved by the Atomic Energy Control Board and has been used to screen 8 patients to date, with 1 patient being denied outpatient treatment. Outpatient therapies with relatively high doses of 131I can be performed safely. Care must be taken to ensure that the patient's home environment is suitable and that the patient can understand and comply with precautions. If external exposure can be minimized, only basic precautions are needed to ensure that internal contamination does not lead to excessive doses to members of the public.


Metal-enhanced fluorescence of an organic fluorophore using gold particles.
Particulate gold films were deposited on glass substrates by vapor deposition. Rabbit immunoglobulin G (IgG) was immobilized by physiosorption and then Alexa Fluor anti Rabbit IgG was bound to the protein-coated surfaces. Fluorescence was enhanced with increasing the Au thickness and reached saturation at 30 nm when Alexa Fluor555 anti IgG was used. We also examined the effect of silica spacers between the gold film and the labeled protein. The maximum enhancement was dependent on the thickness of silica and reach maximum at 10 nm. The maximum increase in intensity was about 6-fold. We also bound Alexa Fluor-680 anti IgG to the protein-coated surface, and the maximum enhancement was about 10-fold.


Self-Assembly of Artificial Sweetener Aspartame Yields Amyloid-like Cytotoxic Nanostructures.
Recent reports have revealed the intrinsic propensity of single aromatic metabolites to undergo self-assembly and form nanostructures of amyloid nature. Hence, identifying whether aspartame, a universally consumed artificial sweetener, is inherently aggregation prone becomes an important area of investigation. Although the reports on aspartame-linked side effects describe a multitude of metabolic disorders, the mechanistic understanding of such destructive effects is largely mysterious. Since aromaticity, an aggregation-promoting factor, is intrinsic to aspartame's chemistry, it is important to know whether aspartame can undergo self-association and if such a property can predispose any cytotoxicity to biological systems. Our study finds that aspartame molecules, under mimicked physiological conditions, undergo a spontaneous self-assembly process yielding regular β-sheet-like cytotoxic nanofibrils of amyloid nature. The resultant aspartame fibrils were found to trigger amyloid cross-seeding and become a toxic aggregation trap for globular proteins, Aβ peptides, and aromatic metabolites that convert native structures to β-sheet-like fibrils. Aspartame fibrils were also found to induce hemolysis, causing DNA damage resulting in both apoptosis and necrosis-mediated cell death. Specific spatial arrangement between aspartame molecules is predicted to form a regular amyloid-like architecture with a sticky exterior that is capable of promoting viable H-bonds, electrostatic interactions, and hydrophobic contacts with biomolecules, leading to the onset of protein aggregation and cell death. Results reveal that the aspartame molecule is inherently amyloidogenic, and the self-assembly of aspartame becomes a toxic trap for proteins and cells, exposing the bitter side of such a ubiquitously used artificial sweetener.


The role of peroxisome proliferator-activated receptor gamma in colon cancer and inflammatory bowel disease.
Review the role and therapeutic potential of peroxisome proliferator-activated receptor (PPAR) gamma in colonic disorders. Recent peer-reviewed scientific literature focusing on PPAR gamma in the colon. Research reports using animal models, cultured cell lines, and clinical material were examined for content related to the role of PPAR gamma in normal colon cell function, colon cancer, and inflammatory bowel disease. Issues concerned with potential therapeutic use were also considered. Key points pertaining to PPAR function and involvement in colon pathology were extracted and noted. Potential compromises to therapeutic utility are identified. The emerging important role of PPAR gamma in normal tissue homeostasis and pathologic outcomes suggests this receptor is a good candidate as a drug target. Several potential problems with this approach will require further investigation prior to widespread recommendations for modulation of PPAR as an efficacious therapy for cancer, chemoprevention of colon cancer, or treatment of inflammatory bowel disease. The widespread use of PPAR gamma ligands for management of type 2 diabetes (such as the glitazone class of drugs including rosiglitazone and pioglitazone) may provide a fortuitous assessment of the efficacy of long-term PPAR modulation.


Preliminary release inventories of unintentionally generated dl-PCB and HCB from sources in China: Base year 2015.
This research presents release inventories of unintentionally generated dioxin-like polychlorinated biphenyls (dl-PCB) and hexachlorobenzene (HCB), which so far have not been developed or assessed. For the inventory development, the amended Toolkit methodology as developed for reporting under the Stockholm Convention on Persistent Organic Pollutants, has been applied. Based on the activity rate (AR) obtained from various statistical yearbooks, reports, or scientific literature, and the emission factors (EFs) suited to each technology level, we estimated the preliminary release inventories of unintentionally produced dl-PCB and HCB from 36 source categories in China for the reference year 2015. The result showed that in 2015, 656 g TEQ of unintentionally produced dl-PCB and 2,145,504 g (or 2146 kg) of unintentionally generated HCB were released in China from these 36 source categories. Most of dl-PCB and HCB was released to air, 71% or 60%, respectively. For comparison - and for the sources which could be estimated for all three unintentional POPs - the total releases of PCDD/PCDF were 5695 g TEQ per year. Of these, 78% were released to air. For dl-PCB and HCB, the vast majority of the releases from the source group 7 - Production and use of consumer goods - is found in the products and not in air. With respect to source attribution and releases to air, the source groups SG3 - Heat and power generation and SG2 - Ferrous and non-ferrous metal production dominate the air emission inventory. Due to the lack of EFs, the calculated releases to the water and land were not well covered, which overall results in an underestimation of the total releases for all unintentional POPs. For comparison, the release inventories from Japan and from several other countries that were developed using methodologies other than the UNEP Toolkit are presented as well.


Editor-in-Chief's Picks From 2014: Part One.
As I spent countless hours pouring over hundreds of manuscripts to select those that rose to the top over the past year, I became incredibly excited about being part of a Journal that produces such wonderfully rich and diverse content each year. I have personally selected the papers (both original investigations and review articles) from 13 distinct specialties for your review. There are approximately 150 articles selected across this 2-part series, which represents less than 3% of the papers submitted to JACC in 2014. In order to present the full breadth of this important research in a consumable fashion, we will present these manuscripts over the course of 2 issues of JACC. Part One includes the sections: Congenital Heart Disease, Coronary Disease & Interventions, Genetics, Omics, & Tissue Regeneration, CV Prevention & Health Promotion, Cardiac Failure, and Cardiomyopathies (1-70). Part Two includes the sections: Hypertension, Imaging, Metabolic Disorders & Lipids, Neurovascular & Neurodegenerative Disorders, Rhythm Disorders, Valvular Heart Disease, and Vascular Medicine.