In the carotid body, galanin is a signal for neurogenesis in young, and for neurodegeneration in the old and in drug-addicted subjects.
The carotid body is a highly specialized chemoreceptive structure for the detection of and reaction to hypoxia, through induction of an increase in hypoxia inducible factor. As tissue hypoxia increases with aging and can have dramatic effects in respiratory depression induced by drug addiction, we investigated the carotid body in young and old healthy subjects in comparison with drug-addicted subjects, including the expression of the neurotransmitter galanin. Galanin expression was recently reported for neuronal-like cells of the human carotid body, and it is implicated in several functions in neurons. In particular, this includes the regulation of differentiation of neural stem cells, and participation in the development and plasticity of the nervous system. Using immunohistochemistry detection, we demonstrate that galanin expression in the human carotid body in healthy older subjects and drug-addicted subjects is significantly reduced in comparison with healthy young subjects. This demonstrates not only the effects of normal aging and senescence, but also in the drug-addicted subjects, this appears to be due to a disorganization of the chemo-sensory region. With both aging and drug addiction, this results in a physiological reduction in neuronal-like cells, coupled with interlobular and intralobular increases in connective tissue fibers. Consequently, in both aging and drug addiction, this reduction of neuronal-like cells and the regeneration suggest that the carotid body is losing its sensory capabilities, with the transmission of chemoreceptive signals dramatically and vitally reduced. The level of galanin expression would thus provide a signal for neurogenesis in young subjects, and for neurodegeneration in older and drug-addicted subjects.


Preparation and Characterization of Stable Nanosuspension for Dissolution Rate Enhancement of Furosemide: A Quality by Design (QbD) Approach.
Nano drug delivery systems have the potential to address the challenges of delivering BCS Class II and IV drugs like furosemide. The purpose of the current study is to prepare stable nanosuspension and investigate in vitro dissolution performance of the model compound furosemide using quality by design (QbD) approach. Nanosuspension batches with uniform particle size were prepared for furosemide using the antisolvent precipitation method. A quality by design (Qbd) approach was explored to understand the impact of process parameters (stirring time, stirring speed, temperature, and injection rate) and material attributes (drug concentration, stabilizer type, drug: stabilizer ratio, and antisolvent: solvent ratio) on the quality attributes of furosemide nanosuspension using a full factorial experimental design. Multiple linear regression and ANOVA were employed to estimate and identify the critical process parameters and material attributes. Injection rate and stirring time were identified as the most critical process parameters' affecting the quality attributes of furosemide nanosuspension. Individual material attributes did not show significant impact on the average particle size of the nanocrystals, however two-way interactions between material attributes (stabilizer type/drug concentration and stabilizer type/antisolvent: solvent ratio) significantly affected nanosuspension particle size distribution. Solid state characterization (PXRD, DSC and SEM) did not exhibit any changes of physical form during preparation and optimization of the furosemide nanosuspension. Dissolution of the furosemide nanocrystals in gastric media was significantly higher than that observed for micronized furosemide suspension and raw furosemide powder. Stability study data suggests that optimized batches of furosemide nanosuspensions were stable for three months at 4°C and ambient conditions. The antisolvent precipitation method can produce stable furosemide nanosuspensions with desirable quality attributes and enhancement of dissolution rate in the gastric medium as compared to the raw furosemide powder and microsuspension.


NOX4 expression in human microglia leads to constitutive generation of reactive oxygen species and to constitutive IL-6 expression.
Reactive oxygen species (ROS) generation by microglia is implicated in neuroinflammation and neurotoxicity, as well as in host defense, cell proliferation and excitatory amino acid release. Recent studies demonstrate that primary microglia preparations not only express the phagocyte NADPH oxidase NOX2, but also the NOX1 and NOX4 isoforms. Here we investigated the relationship between neuroinflammation and NOX isoform expression in the human microglia cell line clone 3 (HMC3). HMC3 cells are typical microglia, as suggested by the constitutive expression of Iba-1 and CD14, and IFN-gamma-induced expression of CD11b, CD68 and MHCII. However, the characteristics of NOX isoform expression and ROS generation by HMC3 cells were unexpected. RT-PCR demonstrated abundant expression of NOX4, but almost no NOX2 mRNA. ROS generation was constitutive and appeared predominantly intracellular, as superoxide was detected within intracellular vesicles, while the cell-permeable H(2)O(2) was found in the extracellular space. ROS generation by HMC3 was efficiently suppressed by siRNA directed against NOX4, but not by control siRNA. NOX4 suppression did not alter expression of the microglia-typical genes MHCII, CD68 and CD11b, nor did it affect the expression of iNOS, VEGF or TGF-beta. However, there was a marked decrease in IL-6 mRNA. Taken together, we demonstrate a constitutive NOX4-dependent ROS generation in a microglial cell line which leads to expression of IL-6 mRNA. The possibility that microglia could switch from tightly regulated NOX2-dependent ROS generation to constitutive NOX4-dependent ROS generation is of interest for the understanding of the role of microglia in maintaining the balance between neuroprotection and neuroinflammatory damage.


Pi-systems as simultaneous hydride and hydrogen bond acceptors.
A theoretical study of the hydride bond complexes with tetrafluoro- and tetracyanoethylene, C2F4 and C2(CN)4, has been carried out by means of density functional theory (DFT) and ab initio methods, up to the MP2/aug-cc-pVTZ computational level. In addition, the ternary complexes formed by an additional standard hydrogen bond donor, such as hydrogen fluoride, have been explored. The results show that the hydride bond complexes are stable and an electron transfer took place from the hydride to the C2F4 and C2(CN)4 molecules. While these molecules are not able to form stable complexes between the pi-electrons and hydrogen bond donors, the presence of the hydrides in the opposite face of the pi-system of C2F4 stabilizes the ternary complexes showing cooperativity effects.


A Concept Analysis of Parental Uncertainty in Illness of an Infant.
The concept of uncertainty in illness has been well described and applied to many different areas of nursing and other disciplines. Specifically, parental uncertainty in illness of an infant is a meaningful concept that has specific attributes and implications. A current concept analysis that considers the changing healthcare setting, historical conceptual inconsistencies, and a lack of information concerning parents of infants is needed. To identify essential antecedents, attributes, and consequences of parental uncertainty in illness using Rodgers' Evolutionary Concept Analysis method. A literature search was conducted using PubMed, CINAHL, and PsycInfo. The following keywords were used in combination using the Boolean terms "AND" and "OR": parental uncertainty; infants; parental uncertainty in illness; preterm infants; parent; uncertainty. articles published between 2000 and 2017 and published in English. The search included 38 articles published from 2000 to 2017 with a specific focus on parental uncertainty in illness. Parental uncertainty in illness of an infant is a paradoxical, cognitive, and emotional experience in which there is an inability to create meaning and may cause disruption in parental role development. Nursing care of parents with ill infants and children must include sensitivity to parents' experiences of uncertainty in illness. Nurses are uniquely positioned to normalize parental uncertainty and facilitate healthy coping.


TEM-1 beta-lactamase as a scaffold for protein recognition and assay.
A large number of different proteins or protein domains have been investigated as possible scaffolds to engineer antibody-like molecules. We have previously shown that the TEM-1 beta-lactamase can accommodate insertions of random sequences in two loops surrounding its active site without compromising its activity. From the libraries that were generated, active enzymes binding with high affinities to monoclonal antibodies raised against prostate-specific antigen, a protein unrelated to beta-lactamase, could be isolated. Antibody binding was shown to affect markedly the enzyme activity. As a consequence, these enzymes have the potential to be used as signaling molecules in direct or competitive homogeneous immunoassay. Preliminary results showed that beta-lactamase clones binding to streptavidin could also be isolated, indicating that some enzymes in the libraries have the ability to recognize proteins other than antibodies. In this paper, we show that, in addition to beta-lactamases binding to streptavidin, beta-lactamase clones binding to horse spleen ferritin and beta-galactosidase could be isolated. Affinity maturation of a clone binding to ferritin allowed obtaining beta-lactamases with affinities comprised between 10 and 20 nM (Kd) for the protein. Contrary to what was observed for beta-lactamases issued from selections on antibodies, enzyme complexation induced only a modest effect on enzyme activity, in the three cases studied. This kind of enzyme could prove useful in replacement of enzyme-conjugated antibodies in enzyme-linked immunosorbant assays (ELISA) or in other applications that use antibodies conjugated to an enzyme.


Premarital genetic screening for beta thalassemia carrier status of indexed families using HbA2 electrophoresis.
To devise a strategy for prevention of beta thalassemia in newborns through reliable screening of indexed families. The cross-sectional study was conducted over six months in 2011 and comprised blood samples collected from subjects belonging to different ethnic groups from families of beta thalassemia major children registered with the Abbottonian Medical Association Blood Care Centre, Abbottabad, in Pakistan's Khyber Pakhtunkhwa province. Electrophoretic separation of human haemoglobin like A, F, S and C was done and then haemoglobin in the gel was immobilised in a fixative solution and the gel was dried to a film. Haemoglobin pattern was visualised by staining the film with a protein-specific stain. The pattern was quantified by densitometry. Of the 98 samples, 57(58.2%) had b-thalassemia trait with elevated haemoglobin alpha 2 level, and 41(41.8%) had normal level. Out of the 57 carriers, 33(57.89%) were males and 24(42.10%) were females. Mean age of carriers was 11.65±6.25 years compared to 10.93±7.75 in normal patients. Mean haemoglobin alpha 2 level of carriers was 5.2±0.56% compared to 2.34±0.57% in normal subjects. Carrying out mass screening programmes throughout Pakistan for the detection of thalassemia carriers and providing them the benefit of marriage counselling may decrease the incidence of thalassemia Major.


Characteristics of opioid receptors in guinea-pig cerebellum and human placenta.
Characteristics of opioid receptors were examined in guinea-pig cerebellum and human placental membrane preparations. Two binding sites for [3H]-ethylketocyclazocine ([3H]-EKCZ) were found in guinea-pig cerebellum membranes whereas only one binding site was detected in human placental membranes. The potencies of kappa opioid agonists were decreased in competing with the binding of [3H]-naloxone to guinea-pig cerebellum membranes or human placental membranes by 5'-guanylylimidodiphosphate (Gpp(NH)P) and sodium. These results suggest that there may be true kappa receptors in human placenta and that kappa receptors in both preparations may couple with the adenylate cyclase system.


Formin-mediated actin polymerization promotes Salmonella invasion.
Salmonella invade host cells using Type 3 secreted effectors, which modulate host cellular targets to promote actin rearrangements at the cell surface that drive bacterial uptake. The Arp2/3 complex contributes to Salmonella invasion but is not essential, indicating other actin regulatory factors are involved. Here, we show a novel role for FHOD1, a formin family member, in Salmonella invasion. FHOD1 and Arp2/3 occupy distinct microdomains at the invasion site and control distinct aspects of membrane protrusion formation. FHOD1 is phosphorylated during infection and this modification is required for promoting bacterial uptake by host cells. ROCK II, but not ROCK I, is recruited to the invasion site and is required for FHOD1 phosphorylation and for Salmonella invasion. Together, our studies revealan important phospho-dependent FHOD1 actin polymerization pathway in Salmonella invasion.


Outcomes and Patient Satisfaction With Arthroscopic Partial Meniscectomy for Degenerative and Traumatic Tears in Middle-Aged Patients With No or Mild Osteoarthritis.
There is controversy about the benefit of arthroscopic partial meniscectomy (APM) for degenerative lesions in middle-aged patients. To compare satisfaction with APM between middle-aged patients with no or mild knee osteoarthritis (OA) and a degenerative meniscal tear and those with a traumatic tear. Cohort study; Level of evidence, 2. A comparative prospective study at 5 years of middle-aged patients (45-60 years old) with no or mild OA undergoing APM for degenerative (n = 115) or traumatic (n = 143) tears was conducted. Patient satisfaction was measured by a 5-point Likert scale and functional outcomes by the Knee injury and Osteoarthritis Outcome Score (KOOS) and Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC). Univariate and multivariate regression analyses were used to identify factors correlating with patient-reported satisfaction at 5 years postoperatively. Baseline patient characteristics were not different between groups. At the 5-year evaluation, the satisfaction rate in the traumatic and degenerative groups was 68.5% versus 71.3%, respectively (P = .365). Patient satisfaction was significantly associated with functional outcomes (r = 0.69; P = .024). In the degenerative group, 43 patients (37.4%) had OA progression to Kellgren-Lawrence (K-L) grade 2 or 3, but only 24 patients (20.8%) had a symptomatic knee at final follow-up. Multivariate regression analysis for patient dissatisfaction at 5-year follow-up showed the following significant independent factors: female sex (odds ratio [OR], 1.6 [95% CI, 1.1-2.3]; P = .018), body mass index >30 kg/m2 (OR, 2.6 [95% CI, 1.7-4.9]; P = .035), lateral meniscal tears (OR, 0.6 [95% CI, 0.1-0.9]; P = .039), and OA progression to K-L grade ≥2 at final follow-up (OR, 1.4 [95% CI, 1.2-2.6]; P = .014). At the final evaluation, there were no significant differences between groups in pain scores (P = .648), WOMAC scores (P = .083), or KOOS-4 scores (P = .187). Likewise, there were no significant differences in the KOOS subscores for Pain (P = .144), Symptoms (P = .097), or Sports/Recreation (P = .150). Although the degenerative group had significantly higher subscores for Activities of Daily Living (P = .001) and Quality of Life (P = .004), the differences were considered not clinically meaningful. There were no meaningful differences in patient satisfaction or clinical outcomes between patients with traumatic and degenerative tears and no or mild OA. Predictors of dissatisfaction with APM were female sex, obesity, and lateral meniscal tears. Our findings suggested that APM was an effective medium-term option to relieve pain and recover function in middle-aged patients with degenerative meniscal tears, without obvious OA, and with failed prior physical therapy.


In situ methane enrichment in anaerobic digestion.
A major cost consideration in the use of anaerobic digestion to convert biomass and waste to utility-grade gas is the expense of separating CO(2) from the product gas. Anaerobic digestion has a number of inherent properties that can be exploited to increase the methane content of the gas directly produced by the digester, the most important of which is the high solubility of CO(2)(40-60 times that of methane) in water under digestion conditions. The methane enrichment concept examined in this study involved the recirculation of a liquid stream from the digester through a CO(2) desorption process and the return of the liquid stream back to the digester for absorption of additional CO(2) produced by the conversion of organic materials. A steady-state equilibrium model predicted that a digester gas methane content exceeding 94% could be achieved with this scheme using modest recirculation rates provided a desorption process could be designed to achieve a 60+% CO(2) removal efficiency in the degassing of the liquid recycle stream. Using fixed-film laboratory digesters operated on synthetic feedstocks, the technique of methane enrichment was tested under pressurized and unpressurized conditions. A 93 + 2% methane gas stream was produced from a volatile-acid-fed bench-scale digester simulating the methanogenic stage of two-phase digestion under conditions of (1) a pH swing achieved without caustic addition that allowed digestion at pH 7. 5 and air stripping at pH 6. 5-7. 0, (2) digester pressurization to 30 psig, and (3) a recycle rate of 0. 33 L/L reactor/day. Significant but lower levels of methane enrichment were achieved with the single-stage digester at the low experimental recycle rate. However, the narrow range among all experiments of CO(2) desorption efficiencies achieved in air stripping the recycle stream (35-60% CO(2) removal) suggests that comparable methane enrichment-may be achieved with unpressurized single-stage digestion using greater recycle rates. A materials balance analysis of data from an unpressurized, single-stage digester employing no chemical addition and using laboratory degassing efficiencies indicated that 94% methane could be produced at recycle rates of less than 1. 4 L/L reactor/day with a methane loss of less than 2%.


Comparison between oral and enteral tube refeeding in hyperlipidemic acute pancreatitis.
Hyperlipidemic acute pancreatitis (HLAP) remains one of the major digestive emergencies with increasing health risks. Oral refeeding tolerant (ORT) and enteral tube feeding tolerant (ETFT) are commonly used for nutritional management in HLAP. However, the differences between ORT and ETFT are yet to be characterized. This study included consecutive patients admitted to the Ordos Central Hospital between January 2019 and April 2023, with predefined inclusion criteria. A total of 335 HLAP patients were recruited according to the inclusion criteria. 268 patients were diagnosed with moderately severe acute pancreatitis (MSAP), of which 193 were in the OFT group and 75 in the ETFT group. In the ETFT group, abdominal pain and abdominal distension were significantly higher than that in the OFT group. No significant result was identified in the laboratory data. However, the OFT group showed a higher hospitalization and cost, as well as exocrine insufficiency and newly onset diabetes, than the ETFT group. Based on the incidence of HLAP retrieved in this study, MSAP is the major type with increasing clinical value. From the nutritional management sense, patients who received OFT showed higher hospitalization and cost, as well as lower exocrine insufficiency and newly onset diabetes.


Loss of renal medullary endothelin B receptor function during salt deprivation is regulated by angiotensin II.
We have recently demonstrated that chronic infusion of exogenous ANG II, which induces blood pressure elevation, attenuates renal medullary endothelin B (ET(B)) receptor function in rats. Moreover, this was associated with a reduction of ET(B) receptor expression in the renal inner medulla. The aim of this present work was to investigate the effect of a physiological increase in endogenous ANG II (low-salt diet) on the renal ET system, including ET(B) receptor function. We hypothesized that endogenous ANG II reduces renal medullary ET(B) receptor function during low-salt intake. Rats were placed on a low-salt diet (0.01-0.02% NaCl) for 2 wk to allow an increase in endogenous ANG II. In rats on normal-salt chow, the stimulation of renal medullary ET(B) receptor by ET(B) receptor agonist sarafotoxin 6c (S6c) causes an increase in water (3.6 ± 0.4 from baseline vs. 10.5 ± 1.3 μl/min following S6c infusion; P < 0.05) and sodium excretion (0.38 ± 0.06 vs. 1.23 ± 0.17 μmol/min; P < 0.05). The low-salt diet reduced the ET(B)-dependent diuresis (4.5 ± 0.5 vs. 6.1 ± 0.9 μl/min) and natriuresis (0.40 ± 0.11 vs. 0.46 ± 0.12 μmol/min) in response to acute intramedullary infusion of S6c. Chronic treatment with candesartan restored renal medullary ET(B) receptor function; urine flow was 7.1 ± 0.9 vs. 15.9 ± 1.7 μl/min (P < 0.05), and sodium excretion was 0.4 ± 0.1 vs. 1.1 ± 0.1 μmol/min (P < 0.05) before and after intramedullary S6c infusion, respectively. Receptor binding assays determined that the sodium-depleted diet resulted in a similar level of ET(B) receptor binding in renal inner medulla compared with rats on a normal-salt diet. Candesartan reduced renal inner medullary ET(B) receptor binding (1,414 ± 95 vs. 862 ± 50 fmol/mg; P < 0.05). We conclude that endogenous ANG II attenuates renal medullary ET(B) receptor function to conserve sodium during salt deprivation independently of receptor expression.


The TLR3 agonist poly(I:C) targets CD8+ T cells and augments their antigen-specific responses upon their adoptive transfer into naïve recipient mice.
We have recently reported that the toll-like receptor 3 (TLR3) agonist poly(I:C) induces adjuvant effects to post vaccination CD8+ T cells responses through rapid induction of innate mediators, including NK cells, macrophages, dendritic cells (DCs), and inflammatory cytokines. However, whether this TLR3 agonist directly targets CD8+ T cells needs to be carefully investigated. In this study, we found that optimal post vaccination CD8+ T cell responses to ex vivo DC-based vaccination requires triggering of TLR3 signaling pathway in DCs in vitro as well as in the recipient host, indicating a role for other cell types. Real-time PCR analysis revealed that TLRs (TLR1-TLR13) are expressed in purified (>99% pure) CD4+ and CD8+ T cells from C57BL/6 and BALB/c mice, where the magnitude of the expression was strain and cell type dependent. In vitro, treatment of these purified T cells with poly(I:C) modulated the expression of TLRs including TLR3. Furthermore, non-specific and antigen-specific stimulation of CD8+ T cells by phorbol myristate acetate and MHC class I peptide-pulsed splenocytes, respectively, modulated TLR expression in purified CD4+ and CD8+ T cells. Importantly, brief conditioning of purified naïve TCR transgenic OT-1 (CD8+) T cells in vitro with poly(I:C) induced activation of these cells in absence of antigen stimulation. Interestingly, when these in vitro poly(I:C)-conditioned OT-1 cells were adoptively transferred into naïve recipient followed by peptide vaccination, they showed superior expansion and activation to their naïve counterparts. These results suggest that CD8+ T cells can be activated by triggering their TLR3. Furthermore, the data support the notion of direct involvement of TLRs in adaptive immune responses.


[Spatial variations of the mechanical properties of dental enamel and dentin].
Cuts with a Vickers diamond were made in five horizontal planes of the upper third of the crown of human lower premolars. At the same time the depth of entry was registered directly during the cutting with adot screen with a distance of about 150 mu between the dots. The depth of the cuts obtained was determined by optical measurement of breadth. Only in the fissure enamel was a slight hardening in the cervical direction observed. With a 5 p loading of the cutter diamond the reversible (elastic) deformation in the dentine is 0,4 +/- 0,1 micrometer greater than in the enamel. The optically determined cutting depth (irreversible deformation) amounted to 1,2 +/- 0,1 micrometer or 2,1 +/- 0,2 micrometer in enamel and dentine.


Interference of BCR-ABL1 kinase activity with antigen receptor signaling in B cell precursor leukemia cells.
The chromosomal translocation t,(9;22) resulting in the fusion of the BCR and ABL1 genes, represents a recurrent aberration in B cell precursor leukemia cells. Their normal counterparts, B cell precursor cells, are positively selected for survival signals through the antigen receptor, whose expression requires a functional immunoglobulin heavy chain (IGH) gene rearrangement. Unexpectedly, B cell precursor leukemia cells harboring a BCR-ABL1 gene rearrangement do not depend on antigen receptor mediated survival signals. Genes involved in the signaling cascade of the antigen receptor are silenced and in most cases, the dominant tumor clone does not carry a functional IGH gene rearrangement. However, upon inhibition of the BCR-ABL1 kinase activity by STI571, only leukemia cells expressing an antigen receptor are able to survive. Since resistance to STI571 is frequent in the therapy of BCR-ABL1(+) B cell precursor leukemia, antigen receptor signaling may represent a mechanism through which these cells can temporarily evade STI571-induced apoptosis. This may open a time frame, during which leukemia cells acquire secondary transforming events that confer definitive resistance to STI571.


Modeling intra-fractional abdominal configuration changes using breathing motion-corrected radial MRI.
Abdominal organ motions introduce geometric uncertainties to gastrointestinal radiotherapy. This study investigated slow drifting motion induced by changes of internal anatomic organ arrangements using a 3D radial MRI sequence with a scan length of 20 min. Breathing motion and cyclic GI motion were first removed through multi-temporal resolution image reconstruction. Slow drifting motion analysis was performed using an image time series consisting of 72 image volumes with a temporal sampling rate of 17 s. B-spline deformable registration was performed to align image volumes of the time series to a reference volume. The resulting deformation fields were used for motion velocity evaluation and patient-specific motion model construction through principal component analysis (PCA). Geometric uncertainties introduced by slow drifting motion were assessed by Hausdorff distances between unions of organs at risk (OARs) at different motion states and reference OAR contours as well as probabilistic distributions of OARs predicted using the PCA model. Thirteen examinations from 11 patients were included in this study. The averaged motion velocities ranged from 0.8 to 1.9 mm min-1, 0.7 to 1.6 mm min-1, 0.6 to 2.0 mm min-1and 0.7 to 1.4 mm min-1for the small bowel, colon, duodenum and stomach respectively; the averaged Hausdorff distances were 5.6 mm, 5.3 mm, 5.1 mm and 4.6 mm. On average, a margin larger than 4.5 mm was needed to cover a space with OAR occupancy probability higher than 55%. Temporal variations of geometric uncertainties were evaluated by comparing across four 5 min sub-scans extracted from the full scan. Standard deviations of Hausdorff distances across sub-scans were less than 1 mm for most examinations, indicating stability of relative margin estimates from separate time windows. These results suggested slow drifting motion of GI organs is significant and geometric uncertainties introduced by such motion should be accounted for during radiotherapy planning and delivery.


Reflections on 30 years of AIDS.
June 2011 marks the 30th anniversary of the first description of what became known as HIV/AIDS, now one of history's worst pandemics. The basic public health tools of surveillance and epidemiologic investigation helped define the epidemic and led to initial prevention recommendations. Features of the epidemic, including the zoonotic origin of HIV and its spread through global travel, are central to the concept of emerging infectious diseases. As the epidemic expanded into developing countries, new models of global health and new global partnerships developed. Advocacy groups played a major role in mobilizing the response to the epidemic, having human rights as a central theme. Through the commitments of governments and private donors, modern HIV treatment has become available throughout the developing world. Although the end of the epidemic is not yet in sight and many challenges remain, the response has been remarkable and global health has changed for the better.


Intact Dynamic Visual Capture in People With One Eye.
Observing motion in one modality can influence the perceived direction of motion in a second modality (dynamic capture). For example observing a square moving in depth can influence the perception of a sound to increase in loudness. The current study investigates whether people who have lost one eye are susceptible to audiovisual dynamic capture in the depth plane similar to binocular and eye-patched viewing control participants. Partial deprivation of the visual system from the loss of one eye early in life results in changes in the remaining intact senses such as hearing. Linearly expanding or contracting discs were paired with increasing or decreasing tones and participants were asked to indicate the direction of the auditory stimulus. Magnitude of dynamic visual capture was measured in people with one eye compared to eye-patched and binocular viewing controls. People with one eye have the same susceptibility to dynamic visual capture as controls, where they perceived the direction of the auditory signal to be moving in the direction of the incongruent visual signal, despite previously showing a lack of visual dominance for audiovisual cues. This behaviour may be the result of directing attention to the visual modality, their partially deficient sense, in order to gain important information about approaching and receding stimuli which in the former case could be life-threatening. These results contribute to the growing body of research showing that people with one eye display unique accommodations with respect to audiovisual processing that are likely adaptive in each unique sensory situation.


Caspase-mediated specific cleavage of BubR1 is a determinant of mitotic progression.
The fidelity of chromosomal duplication is monitored by cell cycle checkpoints operational during mitosis. One such cell cycle delay is invoked by microtubule-targeting agents such as nocodazole or paclitaxel (Taxol) and is mediated by mitotic checkpoint proteins that include BubR1. Relatively little is known about the regulation of expression and stability of BubR1 (or other checkpoint proteins) and how these factors dictate the durability of the cell cycle delay. We report here that treatment of HeLa cells with spindle-disrupting agents resulted in caspase activation and precipitated the cleavage of BubR1. This mechanism ultimately leads to reduced levels of full-length protein, which are accompanied by abrogation of the mitotic block; the checkpoint abrogation is substantially accelerated by inhibition of de novo protein synthesis. In contrast, inhibition of caspase activity blocked BubR1 degradation and prolonged mitosis. To confirm a direct link between caspase activity and BubR1 protein expression, we identified by site-directed mutagenesis the specific caspase cleavage sites cleaved after exposure to paclitaxel. Surprisingly, BubR1 has two sites of cleavage: primarily at Asp607/Asp610 and secondarily at Asp576/Asp579. BubR1 mutated at both locations (BubR1Delta579Delta610) was resistant to paclitaxel-induced degradation. Expression of BubR1Delta579Delta610 augmented the mitotic delay induced by spindle disruption in transfected cells as well as in clones engineered to inducibly express the mutant protein upon exposure to doxycycline and ultimately led to increased aneuploidy. Underscoring the importance of these caspase cleavage sites, both tetrapeptide motifs are identified in the amino acid sequences of human, mouse, chicken, and Xenopus BubR1. These results are potentially the first to link the control of the stability of a key mitotic checkpoint protein to caspase activation, a regulatory pathway that may be involved in killing defective cells and that has been evolutionarily conserved.


Human Papillomavirus infection and cervical lesions in HIV infected women on antiretroviral treatment in Thailand.
To estimate the prevalence and factors associated with Human Papillomavirus (HPV) infection, HPV genotypes and cytological/histological high-grade (HSIL+/CIN2+) lesions. We conducted a cross-sectional study within a prospective cohort of HIV-infected women on combination antiretroviral therapy (cART). Cervical specimens were collected for cytology and HPV genotyping (Papillocheck®). Any women with High-Risk-HPV (HR-HPV), and/or potentially HR-HPV (pHR-HPV) and/or ASC-US or higher (ASC-US+) lesions were referred for colposcopy. Factors associated with HR-HPV infection and with HSIL+/CIN2+ lesions were investigated using mixed-effects logistic regression models. 829 women were enrolled: median age 40.4 years, on cART for a median of 6.9 years, median CD4 cell-count 536 cells/mm3, and 788 (96%) with HIV-viral load<50copies/mL. Of 214 (26%) infected with HPV: 159 (19%) had ≥1 HR-HPV, of whom 38 (5%) HPV52, 22 (3%) HPV16, 9 (1%) HPV18; 21 (3%) had pHR-HPV, 34 (4%) low risk-HPV infection, and 56 (26%) had multiple genotypes. Younger age, low CD4 cell-counts and low education were independently associated with HR-HPV infection. 72 women (9%) had ASC-US+ and 28 (3%) HSIL+/CIN2+ lesions. HR-HPV infection was independently associated with HSIL+/CIN2+ lesions. The prevalence of HPV infection and of cervical lesions was low. The HPV genotype distribution supports the use of 9-valent vaccine in Thailand.


Variability in an early hominin percussive tradition: the Acheulean versus cultural variation in modern chimpanzee artefacts.
Percussion makes a vital link between the activities of early human ancestors and other animals in tool-use and tool-making. Far more of the early human actions are preserved as archaeology, since the percussion was largely used for making hard tools of stone, rather than for direct access to food. Both primate tools and early hominin tools, however, offer a means to exploring variability in material culture, a strong focus of interest in recent primate studies. This paper charts such variability in the Acheulean, the longest-lasting tool tradition, extant form about 1.7 to about 0.1 Ma, and well known for its characteristic handaxes. The paper concentrates on the African record, although the Acheulean was also known in Europe and Asia. It uses principal components and discriminant analysis to examine the measurements from 66 assemblages (whole toolkits), and from 18 sets of handaxes. Its review of evidence confirms that there is deep-seated pattern in the variation, with variability within a site complex often matching or exceeding that between sites far distant in space and time. Current techniques of study allow comparisons of handaxes far more easily than for other components, stressing a need to develop common practice in measurement and analysis. The data suggest, however, that a higher proportion of traits recurs widely in Acheulean toolkits than in the chimpanzee record.


Molecular and Electronic Structures of Ruthenium Complexes Containing an ONS-Coordinated Open-Shell π Radical and an Oxidative Aromatic Ring Cleavage Reaction.
The coordination chemistry of 2,4-di-tert-butyl-6-[(2-mercaptophenyl)amino]phenol (LONSH3), which was isolated as a diaryl disulfide form, (LONSH2)2, with a Ru ion is disclosed. It was established that the trianionic LONS3- is redox-noninnocent and undergoes oxidation to either a closed-shell singlet (CSS), LONS-, or an open-shell π-radical state, LONS•2-, and the reactivities of the [RuII(LONS•2-)] and [RuII(LONS-)] states are different. The reaction of (LONSH2)2 with [Ru(PPh3)3Cl2] in toluene in the presence of PPh3 affords a ruthenium complex of the type trans-[Ru(LONS)(PPh3)2Cl] (1), while the similar reaction with [Ru(PPh3)3(H)(CO)Cl] yields a LONS•2- complex of ruthenium(II) of the type trans-[RuII(LONS•2-)(PPh3)2(CO)] (2). 1 is a resonance hybrid of the [RuII(LONS-)Cl] and [RuIII(LONS•2-)Cl] states. It is established that 2 incorporating an open-shell π-radical state, [RuII(LONS•2-)(CO)], reacts with an in situ generated superoxide ion and promotes an oxidative aromatic ring cleavage reaction, yielding a α-N-arylimino-ω-ketocarboxylate (LNS2-) complex of the type [RuII(LNS2-)(PPh3)(CO)]2 (4), while 1 having a CSS state, [RuII(LONS-)Cl], is inert in similar conditions. Notably, 2 does not react with O2 molecule but reacts with KO2 in the presence of excess PPh3, affording 4. The redox reaction of (LONSH2)2 with [Ru(PPh3)3Cl2] in ethanol in air is different, leading to the oxidation of LONS to a quinone sulfoxide derivative (LONSO0) as in cis-[RuII(LONSO0)(PPh3)Cl2] (3), via 1 as an intermediate. The molecular and electronic structures of 1-4 were established by single-crystal X-ray crystallography, electron paramagnetic resonance spectroscopy, electrochemical measurements, and density functional theory calculations. 1+ is a resonance hybrid of [RuIII(LONS-)(PPh3)2Cl ↔ RuIV(LONS•2-)(PPh3)2Cl]+ states, 2- is a LONS3- complex of ruthenium(II), [RuII(LONS3-)(PPh3)2(CO)]-, and 2+ is a ruthenium(II) complex of LONS- of the type [RuII(LONS-)(PPh3)2(CO)]+, where 35% diradical character of the LONS- ligand was predicted.


Activated ClC-2 Inhibits p-Akt to Repress Myelination in GDM Newborn Rats.
This study aims to investigate the effect and mechanism of type 2 voltage-gated chloride channel (ClC-2) on myelin development of newborn rats' cerebral white matter with gestational diabetes mellitus (GDM). In this study, GDM model was induced in late pregnant rat model. The alteration of ClC-2 expression in various developmental stages of cerebral white matter with/without being exposed to high glucose was analyzed using RT-PCR, active oxygen detection, TUNEL staining, Western Blot as well as immuno-histochemical staining. Our results showed that ClC-2 mRNA and protein expressions in GDM group were significantly increased in white matter of fetal rats after E18 stage, and elevated the level of TNF-α and iNOS in white matter at P0 and P3 stage of newborn rats. Meanwhile, In GDM group, reactive oxygen species (ROS) levels of the white matter at E18, P0, and P3 stage were significantly higher than control group. Furthermore, the expression level of myelin transcription factor Olig2 at P0 stage and CNPase at P3 stage were strikingly lower than that of the control group. In GDM group, ClC-2 expression in the corpus callosum (CC) and cingulate gyrus (CG) regains, and TUNEL positive cell number were increased at P0 and P3 stage. However, PDGFα positive cell number at P0 stage and CNPase expression at P3 stage were significantly decreased. Caspase-3 was also increased in those white matter regions in GDM group, but p-Akt expression was inhibited. While DIDS (a chloride channel blocker) can reverse these changes. In conclusion, ClC-2 and caspase-3 were induced by GDM, which resulted in apoptosis and myelination inhibition. The effect was caused by repressing PI3K-Akt signaling pathway. Application of ClC-2 inhibitor DIDS showed protective effects on cerebral white matter damage stimulated by high glucose concentration.


Ethnicity predicts viral rebound after travel to the tropics in HIV-infected travelers to the tropics in the Swiss HIV Cohort Study.
The number of HIV-infected individuals from developed countries travelling to tropical and subtropical areas has increased as a result of the clinical and survival benefits of combination antiretroviral therapy. The aim of our study was to describe the traveler population in the SHCS and to determine the frequency of viral rebound in virologically suppressed individuals after a travel episode to the tropics compared to non-travelers. Swiss HIV Cohort Study participants with at least one follow-up visit between 1 January 1989 and 28 February 2015 were eligible for inclusion in the study. The primary outcome was the occurrence of viral rebound (viral load > 200 HIV-1 RNA copies/mL) after a travel episode compared with a nontravel episode in previously suppressed individuals (≤ 200 copies/mL). All virologically suppressed patients contributed multiple travel or nontravel episodes to the analysis. Logistic regression was performed including factors associated with viral rebound. We included 16 635 patients in the study, of whom 6084 (36.5%) had ever travelled to the tropics. Travel frequency increased over time, with travellers showing better HIV parameters than nontravellers [less advanced Centers for Disease Control and Prevention (CDC) stage and higher CD4 count nadir]. Viral rebound was seen in 477 (3.9%) of 12 265 travel episodes and in 5121 (4.5%) of 114 884 nontravel episodes [unadjusted odds ratio (OR) 0.87; 95% confidence interval (CI) 0.78-0.97]. Among these 477 post-travel viral rebounds, 115 had a resistance test performed and 51 (44%) of these showed new resistance mutations. Compared with European and North American patients, the odds for viral rebound were significantly lower in Southeast Asian (OR 0.67; 95% CI 0.51-0.88) and higher in sub-Saharan African (SSA) patients (OR 1.41; 95% CI 1.22-1.62). Travel further increased the odds of viral rebound in SSA patients (OR 2.00; 95% CI 1.53-2.61). Region of origin is the main risk factor for viral rebound rather than travel per se. Pre-travel adherence counselling should focus on patients of SSA origin.


Hypomethylating agent induction therapy followed by hematopoietic cell transplantation is feasible in patients with myelodysplastic syndromes.
Disease remission in patients with myelodysplastic syndromes can be achieved with azanucleosides, which act as pyrimidine analogs and hypomethylating agents. However, despite treatment with azanucleoside induction, patients with myelodysplastic syndromes nearly always relapse. Allogeneic hematopoietic cell transplantation (HCT) can be curative, but it is risky. Given that azanucleosides affect human leukocyte antigen expression and lymphocyte reactivity, we conducted a retrospective study to define the impact of pre-HCT azanucleoside therapy on post-HCT donor chimerism. Patients receiving azanucleoside induction therapy achieved rapid and high levels of donor chimerism post-transplant. Lineage analysis also found rapid donor chimerism of lymphocyte and granulocyte subsets. These data indicate the feasibility of pretransplant azanucleoside therapy in patients who subsequently receive an HCT.


Similarities in murine infection and immune response to Borrelia bissettii and Borrelia burgdorferi sensu stricto.
In 1982, Borrelia burgdorferi sensu stricto (ss) was identified as the aetiological agent of Lyme disease. Since then an increasing number of Borrelia burgdorferi sensu lato (sl) species have been isolated in the United States. To date, many of these species remain understudied despite mounting evidence associating them with human illness. Borrelia bissettii is a spirochaete closely related to B. burgdorferi that has been loosely associated with human illness. Using an experimental murine infection model, we compared the infectivity and humoral immune response with a North American isolate of B. bissettii and B. burgdorferi using culture, molecular and serological methods. The original B. bissettii cultures were unable to infect immunocompetent mice, but were confirmed to be infectious after adaptation in immunodeficient animals. B. bissettii infection resulted in spirochaete burdens similar to B. burgdorferi in skin, heart and bladder whereas significantly lower burdens were observed in the joint tissues. B. bissettii induced an antibody response similar to B. burgdorferi as measured by both immunoblotting and the C6 ELISA. Additionally, this isolate of B. bissettii was sequenced on the Ion Torrent PGM, which successfully identified many genes orthologous to mammalian virulence factors described in B. burgdorferi. Similarities seen between both infections in this well-characterized murine model contribute to our understanding of the potential pathogenic nature of B. bissettii. Infection dynamics of B. bissettii, and especially the induced humoral response, are similar to B. burgdorferi, suggesting this species may contribute to the epidemiology of human borreliosis.


Supramolecular host-inhibited excited-state proton transfer and fluorescence switching of the anti-cancer drug, topotecan.
The effect of cucurbit[7]uril (CB[7]) nano-caging on the photophysical properties, particularly excited-state proton transfer (ESPT) reaction, of an eminent anti-cancer drug, topotecan (TPT), is demonstrated through steady-state and time-resolved fluorescence measurements. TPT in water (pH 6) exists exclusively as the cationic form (C) in the ground state. However, the drug emission mainly comes from the excited-state zwitterionic form (Z*) of TPT, and is attributed to water-assisted ESPT between the 10-hydroxyl group and water, which leads to the transformation of C* to Z* of TPT. In the presence of CB[7], it is found that selective encapsulation of the C form of TPT results in the formation of a 1:1 inclusion complex (CB[7]:TPT), and the ESPT process is inhibited by this encapsulation process. As a result, C* becomes the dominant emitting species in the presence of CB[7] rather than Z*, and fluorescence switching takes place from green to blue. Time-resolved studies also support the existence of CB[7]-encapsulated cationic species as the major emitting species in the presence of the macrocyclic host. Semi-empirical quantum chemical calculations are employed to gain insight into the molecular picture of orientation of TPT in the inclusion complex. It is clearly seen from the optimised structure of 1:1 CB[7]:TPT inclusion complex that both 10-hydroxyl and 9-dimethylaminomethylene groups of TPT lie partly inside the cavity, and thereby inhibit the excited-state transformation of C* to Z* by the ESPT process. Finally, controlled release of the drug is achieved by means of fluorescence switching by introducing NaCl, which is rich in cells, as an external stimulus.


Inverse relationship between carrier mobility and bandgap in graphene.
A frequently stated advantage of gapless graphene is its high carrier mobility. However, when a nonzero bandgap is opened, the mobility drops dramatically. The hardness to achieve high mobility and large on∕off ratio simultaneously limits the development of graphene electronics. To explore the underlying mechanism, we investigated the intrinsic mobility of armchair graphene nanoribbons (AGNRs) under phonon scattering by combining first-principles calculations and a tight-binding analysis. A linear dependence of the effective mass on bandgap was demonstrated to be responsible for the inverse mobility-gap relationship. The deformation-potential constant was found to be determined by the strain dependence of the Fermi energy and the bandgap, resulting in two mobility branches, and is essential for the high mobility of AGNRs. In addition, we showed that the transport polarity of AGNRs can be switched by applying a uniaxial strain.


Aldose reductase inhibitors zopolrestat and ferulic acid alleviate hypertension associated with diabetes: effect on vascular reactivity.
This study investigated the effect of aldose reductase (AR) inhibitors on hypertension in diabetes. Diabetes was induced with streptozotocin, while AR inhibitors zopolrestat and ferulic acid were administered at 2 weeks after streptozotocin treatment and for 6 weeks afterwards. Then, blood pressure (BP) and serum level of glucose were determined. Concentration-response curves for phenylephrine (PE), KCl, and acetylcholine (ACh) were obtained in isolated aorta. In addition, ACh-induced NO and reactive oxygen species (ROS) generation in aorta and histopathology were examined. Compared with the control animals, diabetes increased diastolic and systolic BP. AR inhibitors reduced diastolic BP elevation without affecting the developed hyperglycaemia. Diabetes increased the contractile response of aorta to KCl, and decreased the relaxation response to Ach, while administering AR inhibitors prevented an impaired response to ACh. Incubation of aorta isolated from diabetic animals with AR inhibitors did not affect the impaired relaxation response to ACh. In addition, AR inhibitors negated the impaired Ach-stimulated NO generation seen in aorta isolated from diabetic animals. Furthermore, diabetes was accompanied with marked infiltration of leukocytes in aortic adventitia, endothelial cell pyknosis, and increased ROS formation. AR inhibitors reduced leukocyte infiltration and inhibited endothelial pyknosis and ROS formation. In conclusion, AR inhibitors negate diabetes-evoked hypertension via ameliorating impaired endothelial relaxation and NO production.


Factor structure of the Dementia Mood Assessment Scale in a cohort of community-dwelling elderly.
We examined the factor structure of the 28-item Dementia Mood Assessment Scale (DMAS), an instrument to assess depressive symptoms in older adults with cognitive impairment, in a cohort of 165 community-dwelling elderly with varying degrees of cognitive impairment. Factor analysis using principal components analysis and varimax rotation was performed to explore the presence of subscales and examine construct validity. A five-factor structure involving all 28 items accounting for 63.2% of the variance in the DMAS scores was derived. Factors were named: Depressed Affect, Environmental Interaction, Diurnal Patterns, Agitation/Suspicion, and Somatic Indicators. This factor structure reflects the often differing presentations of depressive symptoms in older adults with varying degrees of cognitive function and establishes the construct validity of the DMAS in this population. We conclude that the DMAS may be used for differentiated clinical assessment of depressive symptoms along major dimensions of depressive illness in this cohort of elderly.


Changes in rat respiratory behavior under varying PCO2 levels and argon content in hyperbaric Ar-O2-CO2 atmospheres.
An anesthetized rat preparation has been used for breathing studies in hyperbaric Ar-O2-CO2 atmospheres in which total saturation pressure was varied in the range 1-19.1 ATA and PCO2 was set at levels of 0.0, 0.05 or 0.10 ATA. The variables monitored were respiration rate and an isotonic measure of diaphragm-twitch amplitude during contracture/relaxation (twitch index). The results indicate that: (1) hyperbaric Ar-O2 atmospheres are able to maintain the animal in stable condition for several hours; (2) elevation in PCO2 level produces enhancements in respiration rate and twitch index that are markedly dependent on the PAr level; (3) successive increases in PAr are able to lower progressively the effects on both breathing parameters produced by either 0.05 or 0.10 ATA of CO2, with full abolition of both effects of inhaled CO2 in the PAr range 14-19 ATA; and (4) the intrinsic ability of hyperbaric argon to abolish respiratory enhancement by CO2 is greater than that of hyperbaric helium. These results have been discussed in terms of possible CNS loci at which hyperbaric argon (or helium) is able to antagonize centrally induced effects from dissolved molecular species derived from inspired CO2, and in relation to the broader task of control of excitatory/inhibitory centers of the CNS during diving by manipulation of the partial pressures of diluent components of breathing-gas mixtures.


Effect of a family-based intervention on electronic media use and body composition among boys aged 8--11 years: a pilot study.
This study measured the effect of a 20-week, family-centered electronic media intervention on electronic media use, body composition (dual-energy X-ray absorptiometry; DXA), physical activity and dietary behaviors in boys. Twenty-two boys were assigned and 21 were analyzed in an experimental or control group. Boys in the experimental group set electronic media use goals and used TV Allowance device and ENUFF software to reduce electronic media use. Data were collected at baseline, 10 weeks and 20 weeks. Interactions were found in daily electronic media use and DXA. At 10 weeks, step counts increased by 543 steps per day in the experimental group and decreased by 340 steps per day in the controls. Steps in both groups were higher at 20 weeks. Meals or snacks eaten while using electronic media decreased in the experimental group only. In conclusion, a family-centered electronic media intervention may reduce electronic media use and contribute to desirable changes in body composition.


Computed microtomography visualization and quantification of mouse ischemic brain lesion by nonionic radio contrast agents.
To explore the possibility of brain imaging by microcomputed tomography (microCT) using x-ray contrasting methods to visualize mouse brain ischemic lesions after middle cerebral artery occlusion (MCAO). Isolated brains were immersed in ionic or nonionic radio contrast agent (RCA) for 5 days and subsequently scanned using microCT scanner. To verify whether ex-vivo microCT brain images can be used to characterize ischemic lesions, they were compared to Nissl stained serial histological sections of the same brains. To verify if brains immersed in RCA may be used afterwards for other methods, subsequent immunofluorescent labeling with anti-NeuN was performed. Nonionic RCA showed better gray to white matter contrast in the brain, and therefore was selected for further studies. MicroCT measurement of ischemic lesion size and cerebral edema significantly correlated with the values determined by Nissl staining (ischemic lesion size: P=0.0005; cerebral edema: P=0.0002). Brain immersion in nonionic RCA did not affect subsequent immunofluorescent analysis and NeuN immunoreactivity. MicroCT method was proven to be suitable for delineation of the ischemic lesion from the non-infarcted tissue, and quantification of lesion volume and cerebral edema.


Significance of normal appearance on endoscopic ultrasonography in the diagnosis of early chronic pancreatitis.
The Rosemont classification (RC) was developed as a consensus-based standard for the diagnosis of chronic pancreatitis (CP) by endoscopic ultrasonography (EUS), however, it is more complicated than the conventional scoring system. We have noticed that in the early stages of CP, it is not unusual to observe pancreas with abnormal appearance coexisting with the areas of normal parenchyma. The aim of this study was to investigate the validity of a "normal" pancreas appearance and to evaluate the usefulness of modified diagnostic criteria in comparison to the traditional EUS criteria and the RC. One hundred and seventy-seven patients who had undergone both EUS and endoscopic retrograde pancreatography (ERP) within 2 months were enrolled in the study, and patients with pancreatic cancer were excluded from the study. ERP findings were used as the gold standard for the diagnosis of CP. The EUS images obtained were classified according to both the RC and our new modified criteria. The latter includes an additional criterion to the modified traditional criteria: fine-reticular pattern (F-RP) was defined as a normal pancreatic parenchyma. We compared the accuracy between the new modified EUS criteria and the RC. (1) Normal or equivocal findings on ERP were obtained for 132 patients; 113 patients had F-RP on EUS. In contrast, F-RP was found in only 6 out of 45 CP cases on ERP (P < 0.0001). (2) We investigated the diagnostic capability of our new criteria for endoscopic retrograde cholangiopancreatography normal/equivocal pancreas compared to the traditional criteria. In cases where fewer than two points were defined as normal, the incidence of normal pancreas was significantly higher based on the new criteria than on the traditional criteria (P = 0.002). (3) No significant differences were found between the new criteria and the RC across all ERP grades. Our new proposed "normal-added EUS criteria" for diagnosing CP was equivalent to the RC.


Study of the effect of Tensiomin (captopril) in chronic heart failure.
Tensiomin (captopril) was tested in 23 patients with chronic heart failure for 7.5 months on the average. In 9 outpatients suffering from congestive cardiomyopathy the effect of Tensiomin as an adjuvant to digitalis, diuretic, vasodilator etc. was evaluated by non-invasive methods. During the 3-month follow-up period the heart rate was decreased and the PEP/LVET ratio was improved. The parameters calculated from the X-ray examination (cardiothoracic index, cardiac volume index) indicated the regression of cardiomegaly. Radioisotopic circulatory examinations (systolic volume index, cardiac output index) indicated an increase in cardiac performance. In addition to the significant changes of these parameters the patients' clinical state was also improved in all cases as assessed according to the NYHA classification. Significant side effects were not observed during the treatment.


Assessment of local and regional strategies to control bacteria levels at beaches with consideration of impacts from climate change.
The objective of this study was to evaluate relationships between local factors (beach geomorphology and management) and regional factors (infrastructure improvements and temperature changes) against levels of fecal indicator bacteria (FIB) at recreational beaches. Data were evaluated for 17 beaches located in Monroe County, Florida (Florida Keys), USA, including an assessment of sanitary infrastructure improvements using equivalent dwelling unit (EDU) connections. Results show that elevated FIB levels were associated with beach geomorphologies characterized by impeded flow and by beaches with lax management policies. The decrease in EDUs not connected coincided with a decrease in the fraction of days when bacteria levels were out of compliance. Multivariate factor analysis also identified beach management practices (presence of homeless and concession stands) as being associated with elevated FIB. Overall, results suggest that communities can utilize beach management strategies and infrastructure improvements to overcome the negative water quality impacts anticipated with climate change.


Exploration of the link between tobacco retailers in school neighborhoods and student smoking.
School smoking bans give officials the authority to provide a smoke-free environment, but enacting policies within the school walls is just one step in comprehensive tobacco prevention among students. It is necessary to investigate factors beyond the school campus and into the neighborhoods that surround schools. The purpose of this study was to explore the relationship between the density of tobacco retailers and the illegal tobacco sales rate within school neighborhoods and smoking behaviors among students. This study utilized secondary data from the baseline of the Youth Tobacco Access Project. Data were collected from 10,662 students attending 21 middle schools and 19 high schools, in addition to 512 tobacco retailers, all within 24 towns in Illinois during 2002. A random-effects regression analysis was performed to assess the relationship between the density of tobacco retailers and illegal tobacco sales rates on current smoking and lifetime smoking prevalence. Schools had a range between 0 and 9 tobacco retailers within their neighborhood with a mean of 2.76 retailers (SD = 2.45). The illegal sales rate varied from 0% to 100%, with a mean of 13%. The density of tobacco retailers was significantly related to the prevalence of ever smoking among students (b = 0.09, t(29) = 2.03, p = .051, OR = 1.10), but not to current smoking (p > .05); the illegal tobacco sales rate was not related to current smoking or lifetime smoking prevalence (p > .05). Results indicate that tobacco retailer density may impact smoking experimentation/initiation.


Transient postictal psychosis associated with a course of ECT.
Postictal psychoses, defined as episodic, time-limited psychiatric disturbances of diverse phenomenology, temporally related to a seizure or cluster of seizures, have been well described in patients with focal and generalized epilepsies. Although similar disturbances have been reported in patients undergoing electroconvulsive therapy (ECT), the syndrome of ECT-associated postictal psychosis has not been well described. We review the literature defining the syndrome of postictal psychosis (PIP) associated with the epilepsies, as well as reports of confusional and manic disturbances meeting accepted criteria for PIP, which have been reported to occur as complications of ECT. In addition, we report a case of delusional and hallucinatory psychosis associated with a course of ECT, also having the characteristics of a PIP. We conclude that time-limited psychiatric disturbances, phenomenologically (and possibly pathophysiologically) similar to the PIP of epilepsy can occasionally occur as complications of ECT, and that they may have confusional, manic, delusional, or hallucinatory symptoms. The implications of these occurrences for the use and continuation of ECT are discussed.


Proof-of-Concept Rapid Diagnostic Test for Onchocerciasis: Exploring Peptide Biomarkers and the Use of Gold Nanoshells as Reporter Nanoparticles.
Three O. volvulus immunogenic peptide sequences recently discovered by peptide microarray were adapted to a lateral flow assay (LFA). The LFA employs gold nanoshells as novel high-contrast reporter nanoparticles and detects a serological response against the 3 peptides, found in OvOC9384, OvOC198, and OvOC5528, respectively. When tested on 118 sera from O. volvulus infected patients and 208 control sera, the LFA was 90%, 63%, and 98% sensitive for each peptide, respectively, and 99-100% specific vs samples from healthy volunteers. Samples of other filarial infections cross-reacted by 7-24%. The sensitivity, specificity, and cross-reactivity values matched those obtained by ELISA with the same sample set. While the exact choice of peptide(s) will require fine-tuning, this work establishes that O. volvulus peptides identified by peptide microarray can be translated into an antibody-based LFA and that gold nanoshells provide the same sensitivity, specificity, and cross-reactivity as the corresponding ELISA assays.


Epidemiologic markers of Clostridium difficile.
A wide range of epidemiologic markers have been identified for Clostridium difficile. These markers are based on phenotypic characteristics of the organism, such as antibiotic resistance, bacteriocin or bacteriophage susceptibility, electrophoretic protein patterns, and immunologic markers. Methods for determining genetic markers include plasmid and DNA restriction endonuclease analysis and ribosomal RNA restriction patterns. These methods have been applied to various degrees in epidemiologic studies and have contributed to a better understanding of C. difficile-associated disease. The method based on the use of [35S]methionine-labeled proteins and the development of an automated scanner with computer analysis made possible the recognition of 15 distinct types of C. difficile, based on the major radiolabeled protein patterns. These proteins were strain-specific and immunogenic, as demonstrated by immunoblotting and DNA restriction endonuclease analysis. Epidemiologic studies revealed cross-infection among patients, nosocomial acquisition, and a direct relation between symptoms and type of C. difficile. The development of DNA probes based on antibiotic resistance genes, toxin A, and cloned specific common antigens is discussed.


Electron microscopy of the stria vascularis and its response to etacrynic acid. A study using electron-dense tracers and extracellular surface markers.
Lanthanum nitrate (La3+), ruthenium red (RR) and tannic acid (TA) have been used to examine the cell coat and permeability pathways in thin sections of the stria vascularis of normal and etacrynic-acid (EA)-treated guinea pigs. The tight junctions around the stria, at both endolymphatic and spiral ligament sides, excluded tracers even when EA-associated oedema was well advanced. La3+ was also unable to enter oedematous stria from the capillaries. A dense fibrillar coat was revealed on the endolymphatic marginal cell surface. This coat was disorganised during EA intoxication. Both RR and TA crossed the apical membrane and produced enhanced contrast in some, but not all, marginal cells in EA-treated animals. The results suggest that EA may affect the structure of the apical membrane of the marginal cells and that there is heterogeneity amongst the marginal cell population. These possibilities are discussed.


Self-administered non-invasive vagus nerve stimulation therapy for severe pharmacoresistant restless legs syndrome: outcomes at 6 months.
Severe pharmacoresistant restless legs syndrome (RLS) is difficult to manage and a source of suffering to patients. We studied the effectiveness at 6 months of an innovative treatment: transauricular vagus nerve stimulation (taVNS) in the left cymba concha in a case series of 15 patients, 53% male, mean (SD) age 62.7 (12.3) years with severe pharmacoresistant RLS (mean [SD] International Restless Legs Rating Scale [IRLS] score of 31.9 [2.9]) at baseline. Following an 8-week non-randomised hospital-based study with eight 1-h sessions of taVNS, patients were trained to administer taVNS at home and were followed up for 6 months. The primary outcome measure was the IRLS score, secondary outcome measures were quality of life, mood disorders using the Hospital Anxiety and Depression scale (HAD) subscales for depression (HADD) and anxiety (HADA). At the 6-month follow-up 13/15 patients continued to use weekly taVNS. Symptom severity decreased (mean [SD] IRLS score 22.2 [9.32] at 6 months, p = 0.0005). Four of the 15 patients had an IRLS score of <20 at 6 months and two an IRLS score of 5. Quality of life significantly improved compared to baseline (mean [SD] score at baseline 49.3 [18.1] versus 65.66 [22.58] at 6 months, p = 0.0005) as did anxiety and depression symptoms (mean [SD] HADA score at baseline 8.9 [5.4] versus 7.53 [4.42] at 6 months, p = 0.029; and HADD score at baseline 5.2 [4.5] versus 4.73 [4.44] at 6 months, p = 0.03). Treatment was well tolerated, and no adverse events were reported. Our case series shows a potential role for self-administered taVNS in patients with severe pharmacoresistant RLS. Randomised controlled trials are needed to confirm the utility of taVNS.


The repertoire of testicular extracellular vesicle cargoes and their involvement in inter-compartmental communication associated with spermatogenesis.
Spermatogenesis is regulated by a complex network of intercellular communication processes. Extracellular vesicles (EVs) are one of the important mediators in intercellular communication. Previous reports have demonstrated the involvement of EVs from the epididymis and prostate in sperm maturation and function. However, the presence of EVs in the testis and their potential involvement in spermatogenesis has not been explored. Here, we have established a testis dissociation protocol that allows the isolation and characterization of testicular EVs. We show that testicular EVs are specifically and efficiently taken up by somatic cells and germ cells, including the spermatozoa in the interstitial space and the seminiferous tubule compartments. We profiled the proteome of testicular EVs and probed the cell types that release them, revealing the potential contributions from the Leydig cells and testicular macrophages. Moreover, we sequenced the small RNA cargoes of testicular EVs and identified sets of small non-coding RNAs that were overlooked in the testis transcriptome. Selected miRNA candidates in testicular EVs were found in sperm RNA payload and demonstrated specific resistance towards ribonuclease A independent of the vesicle membrane. Small molecule inhibition of EV secretion perturbed spermatogenesis via inter-compartmental communication. Together, our study provides a valuable resource on the repertoire of cargoes carried by testicular EVs and uncovers a physiological function of testicular EVs in inter-compartmental communication associated to spermatogenesis.


Substance Use and Related Harms Among Adolescents With and Without Traumatic Brain Injury.
The relationship between self-reported lifetime traumatic brain injury (TBI) and drug and alcohol use and associated harms was examined using an epidemiological sample of Canadian adolescents. Data were derived from a 2011 population-based cross-sectional school survey, which included 6383 Ontario 9th-12th graders who self-completed anonymous self-administered questionnaires in classrooms. Traumatic brain injury was defined as loss of consciousness for at least 5 minutes or a minimum 1-night hospital stay due to symptoms. Relative to high schoolers without a history of TBI, those who acknowledged having a TBI in their lifetime had odds 2 times greater for binge drinking (5+ drinks per occasion in the past 4 weeks), 2.5 times greater for daily cigarette smoking, 2.9 times greater for nonmedical use of prescription drugs, and 2.7 times greater for consuming illegal drug in the past 12 months. Adolescents with a history of TBI had greater odds for experiencing hazardous/harmful drinking (adjusted odds ratio [aOR] = 2.3), cannabis problems (aOR = 2.4), and drug problems (aOR = 2.1), compared with adolescents who were never injured. There are strong and demographically stable associations between TBI and substance use. These associations may not only increase the odds of injury but impair the quality of postinjury recovery.


Associations of plasma fibrinogen and factor VII clotting activity with coronary heart disease and stroke: prospective cohort study from the screening phase of the Thrombosis Prevention Trial.
As with 'conventional' risk factors such as cholesterol and smoking, there is a need for large, long-term prospective studies on hemostatic factors. To investigate the prospective relationship of fibrinogen and factor VII clotting activity (FVIIc) with risk of coronary heart disease (CHD) and stroke in a study with a large number of outcomes over a period of 15 years. A cohort of 22 715 men aged 45-69 years was screened for participation in the Thrombosis Prevention Trial. Men were followed up for fatal and non-fatal CHD and stroke events. There were 1515 CHD events (933 CHD deaths) and 391 strokes (180 stroke deaths). Hazard ratios (HRs) and 95% confidence intervals are expressed per standardized increase in log fibrinogen and log FVIIc, adjusting for age, trial treatment group, conventional CHD risk factors and regression dilution bias. Hazard ratios for fibrinogen were 1.52 (1.37-1.70) for all CHD events, and 1.36 (1.09-1.69) for all strokes. Exclusion of events within the first 10 years showed a persistent association between CHD and fibrinogen, with an adjusted HR of 1.93 (1.42-2.64). The HRs for FVIIc, adjusting for age and trial treatment, were 1.07 (1.01-1.12) for all CHD events and 1.07 (0.97-1.20) for all strokes, and the fully adjusted HRs were, respectively, 0.97 (0.84-1.05) and 1.07 (0.85-1.33). The persisting association between fibrinogen and CHD beyond 10 years may imply a causal effect. There is a small effect of FVIIc on CHD, after adjustment for age and trial treatment, but no association independent of other risk factors.


Structural features controlling the binding of beta-carbolines to the benzodiazepine receptor.
Beta-carbolines are a class of drug which can interact with a high affinity with the benzodiazepine (BDZ) binding site of the GABAA receptor. The present paper, aimed at obtaining a deeper insight into the structure-properties relationships of this class of molecules, reports the crystal structures of four beta-carbolines: ZK93423 (3-carboethoxy-4-methoxymethyl-6-benzyloxy-beta-carboline), ZK91296 (3-carboethoxy-4-methoxymethyl-5-benzyloxy-beta-carboline), FG7142 (N-methyl-3-carbamoyl-beta-carboline) and the low-affinity ligand harmine hydrochloride (1-methyl-7-methoxy-beta-carboline). This set of structural data is completed by the X-ray structures of other carbolines of known biological activity retrieved from the Cambridge Crystallographic Database and by the structures of beta-CCE (3-carboethoxy-beta-carboline), 6-PBC (3-carboethoxy-4-methoxymethyl-6-isopropoxy-beta-carboline), PRCC (3-isopropoxy-beta-carboline) and ZK93426 (3-carboethoxy-4-methyl-5-isopropoxy-beta-carboline), which have been obtained by molecular-mechanics simulations. The structural features of all these molecules have been compared according to the stereochemical model we proposed in 1987. The structural comparison is integrated by the Free-Wilson analysis on 32 beta-carbolines of known binding affinity data.


GEOGRAPHIC DISTRIBUTION OF MITOCHONDRIAL DNA VARIANTS AND THE HISTORICAL BIOGEOGRAPHY OF THE SPOTTED SALAMANDER, AMBYSTOMA MACULATUM.
I analyzed geographic partitioning of mitochondrial DNA (mtDNA) restriction-site variants in the spotted salamander, Ambystoma maculatum. Two highly divergent and geographically separate genetic lineages were identified that differed by a minimum of 19 restriction sites (6% sequence divergence). One of the lineages has a disjunct distribution with very closely related haplotypes occurring in Missouri, Arkansas, North Carolina, and Virginia. The other lineage is found in Michigan, Illinois, and Alabama. The geographic separation of highly divergent mtDNA haplotypes, a pattern that was predicted based on the sedentary nature of these salamanders, is evidence for long-term barriers to gene flow. In contrast, the large-scale disjunction of very similar haplotypes suggests recent, long-distance gene flow and does not match the phylogeographic expectation for a small terrestrial vertebrate. I explain this potential contradiction in the level of importance assigned to gene flow by a scenario in which historical barriers to gene flow account for the two divergent mtDNA assemblages, but stochastic sorting of ancestral polymorphism is responsible for the large-scale geographic disjunction. Ten of 16 populations collected in the Ozark Highlands were fixed for the same haplotype. I attribute this lack of detectable variation to recent colonization of this area, a hypothesis that is supported by paleoecological data and demonstrates the potential benefits of combining data from paleobotany, geology, and other disciplines to reconstruct the historical biogeography of a species.


Expression of H-Y antigen by female mice carrying Sxr.
The minor transplantation antigen H-Y can cause graft rejection and can stimulate the generation of H-2-restricted T cell responses. We have used both responses to type karyotypically abnormal mice for the presence of H-Y antigen, in order to investigate the role of H-Y in sex determination. The mice under scrutiny were Sxr5-carrying females derived by crossing females carrying the T(16;X)16H translocation with Sxr carrying males. These females were fully fertile and were H-Y positive. These results are consistent with the testis determining gene, Tdy, which may or may not be H-Y, having a threshold effect on testis differentiation during embryogenesis. They also show that the presence of H-Y in adult females does not impair reproduction.


Novel Optical Biosensor Based on a Nano-Gold Coated by Schiff Base Doped in Sol/Gel Matrix for Sensitive Screening of Oncomarker CA-125.
The urge for sensitive, facile, minimally invasive, and fast detection method of CA-125, a significant and crucial biomarker in ovarian malignancy, is currently substantial. This paper describes the detailed construction and characterization of a newly designed optical nano-biosensor to detect CA-125 accurately and sensitively. The fabricated sensor consists of a nano-gold thin film doped into a matrix of sol-gel, exhibiting a centered fluorescence band at 423 nm when excited at 340 nm. The quantification of CA-125 relies on its quenching ability of this fluorescence signal. The sensor was challenged to evaluate its sensitivity and specificity in detecting CA-125 present in samples collected from ovarian cancer diagnosed patients and compared to samples from healthy women as a control. Our findings revealed that the developed biosensor had a sensitivity of 97.35% and a specificity of 94.29%. Additionally, a wide linearity range over 2.0-127.0 U mL-1 for CA-125 was achieved with a detection limit of 1.45 U mL-1. Furthermore, the sensor could successfully discriminate samples between healthy and diseased people, which demonstrates its suitability in CA-125 assessment.


NHS activity. Running out of juice.
Productivity, as traditionally measured in the NHS, may have levelled off. The Department of Health has acknowledged productivity problems. Current measurement indicators do not take account of quality, so improvement to services may show up as decreases in efficiency. Quality improvements should be included in productivity measurement.


Preparation and properties of cardiac cytochrome c1.
A method for the large-scale isolation of beef heart mitochondrial cytochrome c1 in high purity was developed. This method gave higher yield of "one-band" cytochrome c1 than previously reported [Kim, C. H., & King, T. E. (1981) Biochem. Biophys. Res. Commun. 102, 607-614]. In addition, the present method was effective in the preparation of "two-band" cytochrome c1 which was used to prepare the hinge protein according to the principle of sequential resolution [Kim, C. H., & King, T. E. (1983) J. Biol. Chem. 258, 13543-13551]. The isolation of one-band and two-band cytochrome c1 by this procedure could be completed within 3 or 4 days starting with succinate-cytochrome c reductase. One-band cytochrome c1 showed a molecular weight of 44,000 by sedimentation equilibrium and 29,000 by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The disparities in these data from the actual value of 27,924 by amino acid sequence analysis, as previously reported [Wakabayashi, S., Matsubara, H., Kim, C. H., & King, T. E. (1982) J. Biol. Chem. 257, 9335-9344], are most probably due to the formation of detergent or detergent-phosphate complex. A comparison of some properties of one-band cytochrome c1 with those of two-band cytochrome c1 clearly showed significant differences between the two preparations. These results suggest the hypothesis that one of the possible roles of the hinge protein in the mitochondrial respiratory chain is to stabilize the conformation of cytochrome c1.


Spinal versus general anesthesia for Cesarean section in patients with sickle cell anemia.
Sickle cell anemia (SCA) increases the rate of maternal and fetal complications. This pilot study was designed to compare the maternal and fetal outcomes of spinal versus general anesthesia (GA) for parturients with SCA undergoing cesarean delivery. Forty parturients with known SCA scheduled for elective Cesarean delivery were randomized into spinal anesthesia (n = 20) and GA groups (n = 20). Perioperative hemodynamic parameters were recorded. Postpartum complications were followed up. Opioid consumption was calculated. Blood loss during surgery and the number of patients who received intraoperative or postpartum blood transfusion were recorded. Patient satisfaction with the type of anesthesia was assessed. The Apgar score at 1 and 5 min, neonatal admission to the intensive care unit, and mortality were also recorded. Blood loss was significantly higher in the GA than spinal group (P = 0.01). However, the number of patients who received an intraoperative or postpartum blood transfusion was statistically insignificant. Significantly more patients developed intraoperative hypotension and bradycardia in the spinal than GA group. Opioid use during the first 24 h was significantly higher in the GA than spinal group (P < 0.0001). More patients had vaso-occlusive crisis in the GA than spinal group without statistical significance (P = 0.4). There was one case of acute chest syndrome in the GA group. No significant differences were observed in postoperative nausea and/or vomiting, patient satisfaction, or hospital length of stay. Neonatal Apgar scores were significantly better in the spinal than GA group at 1 and 5 min (P = 0.006 and P = 0.009, respectively). Neonatal intensive care admission was not significantly different between the two groups, and there was no neonatal mortality. Spinal anesthesia may have advantages over GA in parturients with SCA undergoing Cesarean delivery.


Testing microbiologic response to antiinfective medications with incomplete data.
Clinical trials of antiinfective medications often require estimates of the proportions of patients, n, who are free of disease-causing pathogens at the end of treatment, as well as the proportions of all pathogens that have been eradicated. Each patient is infected with several species of pathogens, but the response to study medication for some of these pathogens may be unknown because some specimens were lost or because the patient received nonstudy medication that was known to be effective against these species. This paper develops a statistical model that estimates pi for each treatment and that accounts for the unknown pathogen responses as well as overdispersion of the remaining responses due to within-patient effects. The data are modeled with the Poisson distribution for the numbers of pathogen species per patient and the beta-binomial model for pathogen eradication. The Poisson and beta-binomial parameters are estimated through maximum likelihood estimation, and the treatment difference in the pi valves and its standard error are estimated by transforming the underlying parameters. Confidence intervals based on these estimates are constructed to test the hypothesis of noninferiority of the test treatment.


Painful oral mucosal ulcers in a patient with small cell carcinoma of the lung.
This case illustrates the development of multiple painful oral ulcers caused by methotrexate that was one of a combination of chemotherapeutic drugs administered for the treatment of small cell carcinoma of the lung. Although the oral mucositis is self-limiting and resolves when the drug dose is reduced or therapy is discontinued, severe pain and discomfort may cause physical debilitation. Moreover, the risk of secondary oral infections is high in patients undergoing such therapy, and if the appropriate treatment is not instituted, fatal systemic dissemination of the infection may occur.


[Analysis of results of stapedotomy in patients with obliterative otosclerosis].
In obliterative type of otosclerosis the stapes footplate is greatly thickened and diffusely replaced by a massive otosclerotic masses that fill in the oval window niche, often annular ligament cannot be visualized. In the case of obliterative otosclerosis the only effective way of treatment is surgery. The analysed group consisted of 53 patients with primary obliterative otosclerosis. In the control group there were 30 consecutive patients treated for otosclerosis. The auditory organ was assessed on the ground of the following criteria for the frequencies 0.5, 1, 2 and 4 kHz.: 1. Change of pure tone thresholds of air conduction (AC) 2. Change of pure tone thresholds of bone conduction (BC) 3. Change of pure tone thresholds of air-bone gap (ABG) as the difference between the mean AC and BC thresholds before the operation (AC0 and BC0) and 12 months after the operation (AC1 and BC1). Postoperatively mean value of change of AC thresholds in the analysed group was about 24 +/- 2 dBHL and in the control group about 25 +/- 2 dBHL (no significant difference). In the analysed group the mean value of BC thresholds varied about 8 dBHL and in the control group about 11 dBHL (no significant difference). The average values of ABG after the operation varied about 14 +/- 1 dBHL in both--the analysed and the control group. Postoperatively the mean value ABG was amounted to 20 +/- 1 dBHL in the analysed group and in the control group to 18 +/- 1 dBHL (no significant difference). On the basis of the subjective assessment completely subsidence of aural tinnitus reported 70% patients in the analysed group and 80% patients in the control group. 1. The incidence of obliterative otosclerosis represents 9,6% of all otosclerotic patients treated surgically. 2. Stapedotomy is the treatment of choice in patients with obliterative otosclerosis, because significant improvement of AC, BC and ABG 12 months after the operation was reported, mean values did not differ between the above and control group. 3. Complete subsidence of aural tinnitus was reported in 70% patients with obliterative otosclerosis.


Comparative genomic analysis of the proteasome β5t subunit gene: implications for the origin and evolution of thymoproteasomes.
The thymoproteasome is a recently discovered, specialized form of 20S proteasomes expressed exclusively in the thymic cortex. Although the precise molecular mechanism by which the thymoproteasome exerts its function remains to be elucidated, accumulating evidence indicates that it plays a crucial role in positive selection of T cells. In the present study, we analyzed the evolution of the β5t subunit, a β-type catalytic subunit uniquely present in thymoproteasomes. The gene coding for the β5t subunit, designated PSMB11, was identified in the cartilaginous fish, the most divergent group of jawed vertebrates compared to the other jawed vertebrates, but not in jawless vertebrates or invertebrates. Interestingly, teleost fish have two copies of apparently functional PSMB11 genes, designated PSMB11a and PSMB11b, that encode β5t subunits with distinct amino acids in the S1 pocket. BLAST searches of genome databases suggest that birds such as chickens, turkey, and zebra finch lost the PSMB11 gene, and have neither thymoproteasomes nor immunoproteasomes. In mammals, reptiles, amphibians, and teleost fishes, the PSMB11 gene (the PSMB11a gene in teleost fish) is located next to the PSMB5 gene coding for the β5 subunit of the standard 20S proteasome, indicating that the PSMB11 gene arose by tandem duplication from the evolutionarily more ancient PSMB5 gene. The general absence of introns in PSMB11 and an unusual exon-intron structure of jawed vertebrate PSMB5 suggest that PSMB5 lost introns and duplicated in tandem in a common ancestor of jawed vertebrates, with PSMB5 subsequently gaining two introns and PSMB11 remaining intronless.


Fragmentation studies on monensin A by sequential electrospray mass spectrometry.
Monensin A was studied by electrospray ionisation sequential mass spectrometry (ESI-MSn) and all fragments were confirmed by accurate-mass measurements. Analyses were performed on both a quadrupole time-of-flight (Q-tof) and a Fourier-transform ion cyclotron resonance (FT-ICR) mass spectrometer. MSn analysis shows that depending on sample preparation the ion at m/z 671 consists of two different ions with the same accurate-mass. It is either the monensin protonated parent ion or a different ion structure derived from the loss of water from the water adduct of monensin. Both ions show different fragmentation patterns. Major fragment ions from the protonated parent ion were produced by Grob-Wharton type fragmentations in addition to various simple neutral losses. The fragmentation pathways of the two different m/z 671 ions are proposed.


Effects of duty cycles on passive acoustic monitoring of southern resident killer whale (Orcinus orca) occurrence and behavior.
Long-term passive acoustic monitoring of cetaceans is frequently limited by the data storage capacity and battery life of the recording system. Duty cycles are a mechanism for subsampling during the recording process that facilitates long-term passive acoustic studies. While duty cycles are often used, there has been little investigation on the impact that this approach has on the ability to answer questions about a species' behavior and occurrence. In this study, the effects of duty cycling on the acoustic detection of southern resident killer whales (SRKW) (Orcinus orca) were investigated. Continuous acoustic data were subsampled to create 288 subsampled datasets with cycle lengths from 5 to 180 min and listening proportions from 1% to 67%. Duty cycles had little effect on the detection of the daily presence of SRKW, especially when using cycle lengths of less than an hour. However, cycle lengths of 15-30 min and listening proportions of at least 33% were required to accurately calculate durations of acoustic bouts and identify those bouts to ecotype. These results show that the optimal duty cycle depends on the scale of the research question and provide a framework for quantitative analysis of duty cycles for other marine species.


Fluorescent Molecular Probes for Detection of One-Electron Oxidants Photochemically Generated by Dissolved Organic Matter.
We report a dual probe system based on 4'-substituted biphenyl-2-carboxylic acids (BPAs) for analysis of photooxidants generated by dissolved organic matter. The BPA probes are converted to the corresponding benzocoumarins (BZCs) at different rates depending on the mechanism of oxidation; thus, two probes used simultaneously can differentiate strong triplet excited state sensitizers from hydroxylating species such as hydroxyl radical (•OH) present in dissolved organic matter (DOM). Comparison of the ratios of BZC-CH3 and BZC-CF3 product formation using model photooxidants such as NaNO2, a •OH precursor, and model triplet sensitizer lumichrome gave a range of 2 to 250. Application of these probes to DOM isolates and whole natural waters afforded intermediate ratios. Although the oxidation potential of BPAs (>ca. 1.80 V SHE) is significantly higher than the estimated average reduction potential of typical 3CDOM* samples, these results have demonstrated the presence of a small pool of oxidants in the selected DOM isolates and whole water samples that is capable of oxidizing aromatic carboxylates. As an analytical tool, this probe pair can be used between pH 4-6 without affecting the product formation ratio and may find applications in various systems involving complex mixtures of photochemically produced oxidants of differing natures.


Getting there: directions given by mildly retarded and nonretarded adults.
We found, not surprisingly, that most nonretarded adults can either give accurate and adequate sets of directions to their places of residence or recognize their inability to do so and refuse to attempt to give any--86% of those in our sample. We found also that this is the case for many mentally retarded adults who reside in the community--45% of those in our sample. These individuals responded in perfectly appropriate ways to a request for directions. However, 55% of the mentally retarded sample gave inaccurate or ineffective directions. Of these, 64% made errors that differed in quantity--both in terms of total number of errors and number of errors per set of directions--but not in kind from the errors made by the nonretarded sample. This leaves 13 mildly mentally retarded individuals--20% of our total sample, 36% of those who gave inaccurate directions--who made errors that were unlike those made by nonretarded adults: errors of communicative competence that involved a lack of knowledge of the structure and function of directions or a lack of ability in communicative design. In our studies of other everyday speech events such as narratives, we have suggested that one of the reasons for the impression interlocutors have that something is different or wrong with the way that mildly mentally retarded persons speak is that although they make the same sorts of errors that nonretarded speakers make, they make them more frequently. Because of the nature of the speech events we have studied, we have been unable to demonstrate this conclusively. However, as we pointed out at the beginning of this paper, the nature of directions is such that one can make objective judgements of accuracy and of type and number of errors. For this speech event at least, our suggestion that mildly mentally retarded speakers make errors that are like those made by nonretarded speakers but make them more frequently, is confirmed. Furthermore, we have found that difficulties with communicative design that are not usually exhibited in the speech of nonretarded persons are present not only in the narratives of mentally retarded speakers but in the directions that they give as well. Traditionally, studies of language and mental retardation have concentrated on subjects' phonological, morphological, lexical, and syntactic deficits. But for the mildly retarded, such deficits are relatively insignificant.(ABSTRACT TRUNCATED AT 400 WORDS)


A concertedly evolving region in Chironomus, unique within the telomere.
Chromosome terminal, complex repeats in the dipteran Chironomus pallidivittatus show rapid concerted evolution during which there is remarkably efficient homogenization of the repeat units within and between chromosome ends. It has been shown previously that gene conversion is likely to be an important component during these changes. The sequence evolution could be a result of different processes-exchanges between repeats in the tandem array as well as information transfer between units in different chromosomes-and is therefore difficult to analyze in detail. In this study the concerted evolution of a region present only once per chromosome, at the junction between the telomeric complex repeats and the subtelomeric DNA was therefore investigated in the two sibling species C. pallidivittatus and C. tentans. Material from individual microdissected chromosome ends was used, as well as clones from bulk genomic DNA. On the telomeric side of the border pronounced species-specific sequence differences were observed, the patterns being similar for clones of different origin within each species. Mutations had been transmitted efficiently between chromosomes also when adjoining, more distally localized DNA showed great differences in sequence, suggesting that gene conversion had taken place. The evolving telomeric region bordered proximally to subtelomeric DNA with high evolutionary constancy. More proximally localized, subtelomeric DNA evolved more rapidly and showed heterogeneity between species and chromosomes.


Could Serum TSH Levels Predict Malignancy in Euthyroid Patients Affected by Thyroid Nodules with Indeterminate Cytology?
Serum TSH levels in the upper-normal range were reported to be associated with increased risk of thyroid malignancy. However, measurement of TSH levels is currently not recommended for assessing the risk of malignancy in patients with newly diagnosed thyroid nodules. To evaluate a possible relationship between the serum levels of TSH and the histological outcome of patients undergoing thyroidectomy for thyroid nodules with indeterminate cytology. We collected the clinical data of all patients who had performed ultrasound-guided FNA of thyroid nodules with cytological diagnosis of indeterminate lesions (TIR3A and TIR3B) and serum TSH levels within the normal range. All patients had been submitted to thyroid surgery (hemi or thyroidectomy, as appropriate), and histological diagnosis had been performed. A histological diagnosis of thyroid malignancy was rendered in 74/378 (19.6%) nodules. Patients with histologically proven thyroid malignancy were characterized by higher serum levels of TSH as compared to patients with histologically proven benign nodules (3.03 ± 1.16 vs. 2.37 ± 1.19 mIU/L, p < 0.001). To further analyze the role of serum TSH in predicting thyroid cancer, patients were stratified in 4 groups according to quartiles of TSH concentrations. The prevalence of malignancy was 12.2% for the first quartile and 50.0% for the last quartile. ROC curve analysis identified that a serum TSH level of ≥2.7 mIU/L predicted thyroid malignancy with a sensitivity of 61% and a specificity of 65%. TSH levels in the upper-normal range are associated with an increased risk of thyroid malignancy in patients affected by thyroid nodules with indeterminate cytology at FNA. The measurement of serum TSH levels represents an easily performed additional tool for decision-making in patients with indeterminate cytological findings.


Enantioselective Synthesis of Hemiaminals via Pd-Catalyzed C-N Coupling with Chiral Bisphosphine Mono-oxides.
A novel approach to hemiaminal synthesis via palladium-catalyzed C-N coupling with chiral bisphosphine mono-oxides is described. This efficient new method exhibits a broad scope, provides a highly efficient synthesis of HCV drug candidate elbasvir, and has been applied to the synthesis of chiral N,N-acetals.


Growth of AM fungi on in vitro root organ culture of Sorghum vulgare and Saccharum officinarum.
Spores of Gl mosseae and Gig gigantea germinated on minimal medium produced extraradical mycelium. Gl. mosseae infected roots of S. officinarum in in vitro condition were inoculated in M medium with in vitro cultured roots of Sorghum vulgare (test roots). From the infected root of S. officinarum, the mycelium developed and it infected the test roots. The roots developed new mycelia and further the mycelia produced a few hyaline spores. In MS medium combined with soil extract, root exudate, thiamine HCl and inositol combination, spore germination and germ tube growth were higher when compared with other media.