Active absorption of NH4+ by rat medullary thick ascending limb: inhibition by potassium.
These experiments were designed to determine the relative contributions of active NH4+ transport and voltage-driven NH4+ diffusion to direct NH4+ absorption by the medullary thick ascending limb of the rat. Medullary thick ascending limbs were perfused in vitro with solutions containing 25 mM HCO3 and 4 mM total ammonia. Under steady-state conditions, the lumen-positive transepithelial voltage (VT) was not sufficient to account for the observed decrease in lumen NH4+ concentration, consistent with active absorption of NH4+. Flux calculations based on VT and measured NH4+ permeability (6 x 10(-5) cm/s) indicate that the majority (at least 65%) of total ammonia absorption is due to active transport of NH4+. The remainder of NH4+ absorption can be accounted for by voltage-driven diffusion. Increasing the potassium concentration from 4 to 24 mM in perfusate and bath markedly inhibited total ammonia absorption but did not affect VT, NH4+ permeability, or HCO3 absorption. These results are consistent with inhibition of the active component of NH4+ absorption by potassium. The active NH4+ absorption is likely mediated by cotransport of Na+, NH4+, and Cl- across the apical cell membrane. Inhibition of active NH4+ absorption by an increase in potassium concentration may be due, in part, to competition between NH4+ and K+ for a common binding site on the Na+ -K+ -2Cl- cotransport system.


Interfacial structure and orientation of confined ionic liquids on charged quartz surfaces.
Atomistic molecular dynamics simulations have been performed to study microscopic ionic structures and orientational preferences of absorbed [BMIM] cations and four paired anions ([BF4], [PF6], [TFO] and [TF2N]) on quartz surfaces. Two chemically different quartz surface models were adopted: one is saturated with silanol Si(OH)2 groups, and the other one is covered by silane SiH2 groups, respectively. Simulation results reveal that dense ionic layers, characterized by distinct mass, number, charge and electron densities, are formed in quartz interfacial region. The orientational preferences of confined ionic groups are characterized with different features depending on the size and shape of anionic groups, and the quartz surface charge. The [BMIM] cations attach exclusively onto the negatively charged Si(OH)2 surface. The imidazolium rings lie preferentially perpendicular to Si(OH)2 surface, to which the directly connected methyl and butyl chains are oriented and elongated along Si(OH)2 surface, respectively. The anions are mainly absorbed on positively charged SiH2 surface. The main axes of asymmetric [TFO] and [TF2N] anions are perpendicular and parallel to SiH2 surface, respectively. Such distinct structural and orientational preferences of confined ionic groups attribute to the strong electrostatic interactions and the formation of hydrogen bonds between confined ionic species and quartz interfacial groups.


Genome-wide analysis in chicken reveals that local levels of genetic diversity are mainly governed by the rate of recombination.
Polymorphism is key to the evolutionary potential of populations. Understanding which factors shape levels of genetic diversity within genomes forms a central question in evolutionary genomics and is of importance for the possibility to infer episodes of adaptive evolution from signs of reduced diversity. There is an on-going debate on the relative role of mutation and selection in governing diversity levels. This question is also related to the role of recombination because recombination is expected to indirectly affect polymorphism via the efficacy of selection. Moreover, recombination might itself be mutagenic and thereby assert a direct effect on diversity levels. We used whole-genome re-sequencing data from domestic chicken (broiler and layer breeds) and its wild ancestor (the red jungle fowl) to study the relationship between genetic diversity and several genomic parameters. We found that recombination rate had the largest effect on local levels of nucleotide diversity. The fact that divergence (a proxy for mutation rate) and recombination rate were negatively correlated argues against a mutagenic role of recombination. Furthermore, divergence had limited influence on polymorphism. Overall, our results are consistent with a selection model, in which regions within a short distance from loci under selection show reduced polymorphism levels. This conclusion lends further support from the observations of strong correlations between intergenic levels of diversity and diversity at synonymous as well as non-synonymous sites. Our results also demonstrate differences between the two domestic breeds and red jungle fowl, where the domestic breeds show a stronger relationship between intergenic diversity levels and diversity at synonymous and non-synonymous sites. This finding, together with overall lower diversity levels in domesticates compared to red jungle fowl, seem attributable to artificial selection during domestication.


Purification and characterization of Staphylococcus aureus type 8 capsular polysaccharide.
Clinical isolates of Staphylococcus aureus have been previously classified into eight types on the basis of their capsular polysaccharide. The high prevalence of the type 8 capsular polysaccharide among bacteremic isolates suggests the importance of this capsular antigen in staphylococcal disease. The capsular polysaccharide was purified from extracts of three clinical isolates of S. aureus type 8 of different geographic and temperal origin by ion-exchange chromatography and gel filtration. Gas chromatography, gas chromatography-mass spectrometry, and 13C-nuclear magnetic resonance showed that the type 8 capsular polysaccharide is composed of O-acetyl groups, N-acetylfucosamine, and an aminouronic acid similar to N-acetylgalactosaminouronic acid. The purified polysaccharide reacted only with type 8 antiserum in double diffusion experiments. Our analysis shows that the type 8 polysaccharide is both chemically and serologically distinct from teichoic acid and previously characterized polysaccharides of S. aureus.


Variables affecting estimated glomerular filtration rate after renal transplantation in children: a NAPRTCS data analysis.
Short-term graft survival has improved in renal transplants without significant effect on long-term graft survival. As GFR decline precedes graft loss, an understanding of variables affecting eGFR after TX may help improve graft survival. NAPRTCS data were analyzed to assess effects of donor, recipient, and other variables on Schwartz eGFR after transplantation. For 8438 children with a functioning graft at day 30, data were censored for children dying with a functioning graft, and those with <3 yr follow-up. Multivariate linear regression and repeated measures analyses identified factors related to eGFR at day 30 after TX and during follow-up. Young, female, non-black, children without ATN and acute rejection in the first 30 days, TX after 1995, those with better eGFR at day 30, and receiving tacrolimus had better long-term eGFR. Transplant from ideal (6-35 yr) donors had best short-term eGFR, young donors (<5 yr) had lower eGFR and poor graft survival. After one yr, eGFR improved in surviving grafts of young donors and matched ideal donors. Acute rejection, BP medications, and hospitalizations in prior six months had negative association with subsequent eGFR. Regardless of variables, eGFR deteriorated with time. Slope of eGFR decline has not changed in the recent era indicating the need for innovative therapies.


Use of Antihypertensive Medications and Risk of Adverse Breast Cancer Outcomes in a SEER-Medicare Population.
Background: It is unclear if use of common antihypertensive medications influences the risk of adverse breast cancer outcomes.Methods: Using the linked Surveillance, Epidemiology and End-Results (SEER)-Medicare database, we identified 14,766 women between ages 66 and 80 years diagnosed with incident stage I/II breast cancer between 2007 and 2011. Medicare Part D data were obtained to characterize women's post-cancer use of various antihypertensive medications. Outcomes included a second breast cancer event (SBCE; a composite outcome defined as the first of a recurrence or a second contralateral primary breast cancer), breast cancer recurrence, and breast cancer-specific mortality. Time-varying Cox proportional hazard models were used to estimate hazard ratios (HR) and their associated 95% confidence intervals (CI).Results: There were 791 SBCEs, 627 breast cancer recurrences, and 237 breast cancer deaths identified over a median follow-up of 3 years. Use of diuretics (n = 8,517) after breast cancer diagnosis was associated with 29% (95% CI, 1.10-1.51), 36% (95% CI, 1.14-1.63) and 51% (95% CI, 1.11-2.04) higher risks of a SBCE, recurrence, and breast cancer death, respectively. Compared with nonusers, β-blockers users (n = 7,145) had a 41% (95% CI, 1.07-1.84) higher risk of breast cancer death. Use of angiotensin II receptor blockers, calcium channel blockers and angiotensin-converting enzyme inhibitors were not associated with risks of breast cancer outcomes.Conclusions: Use of diuretics and β-blockers may be associated with increased risk of breast cancer outcomes among older women.Impact: Most antihypertensive medications are safe with respect to breast cancer outcomes, but more research is needed for diuretics and β-blockers. Cancer Epidemiol Biomarkers Prev; 26(11); 1603-10. ©2017 AACR.


Evaluation of a New Histoplasma spp. Quantitative RT-PCR Assay.
Laboratory diagnosis of histoplasmosis is based on various methods, including microscopy, culture, antigen, and DNA detection of Histoplasma capsulatum var. capsulatum or Histoplasma capsulatum var. duboisii. To improve sensitivity of existing real-time quantitative PCR (qPCR) assays, we developed a new RT-qPCR assay that allows amplification of whole nucleic acids of Histoplasma spp. validated on suspected cases. The limit of detection was 20 copies, and the specificity against 114 fungal isolates/species was restricted to Histoplasma spp. Whole nucleic acids of 1319 prospectively collected consecutive samples from 907 patients suspected of having histoplasmosis were tested routinely between May 2015 and May 2019 in parallel with standard diagnostic procedures performed in parallel. Forty-four had proven histoplasmosis attributable to H. capsulatum var. capsulatum (n = 40) or H. capsulatum var. duboisii (n = 4) infections. The results of RT-qPCR were positive in 43 of 44 patients (97.7% sensitivity) in at least one specimen. Nine of 863 cases (99% specificity) were RT-qPCR positive and therefore classified as possible cases. RT-qPCR was positive in 13 of 30 (43.3%) blood samples tested in proven cases. A positive RT-qPCR result in blood was significantly associated with H. capsulatum var. capsulatum progressively disseminated histoplasmosis with a positive RT-qPCR result in 92.3% of the immunocompromised patients with disseminated disease. This new Histoplasma RT-qPCR assay enabling amplification of H. capsulatum var. capsulatum and H. capsulatum var. duboisii is highly sensitive and allows the diagnosis of histoplasmosis advantageously from blood and bronchoalveolar lavage fluid.


Reduction of noise-induced hearing loss using L-NAC and salicylate in the chinchilla.
The effects of a combination of two antioxidant compounds were studied in a chinchilla model of noise-induced hearing loss. After obtaining baseline hearing thresholds using inferior colliculus evoked potentials, chinchillas were exposed for 6 h to octave band noise centered at 4 kHz (105 dB SPL). Post-noise thresholds were obtained 1 h after the noise exposure, and then animals received either saline or salicylate and N-L-acetylcysteine combination. Another group received antioxidant treatment 1 h prior to noise. Hearing was tested at 1, 2 and 3 weeks post-noise. Subsequently, the cochleae were harvested, and cytocochleograms were prepared. There was a 20-40 dB SPL threshold shift at 3 weeks for tested controls. Permanent threshold shifts (PTS) were significantly reduced (P<0.05) to approximately 10 dB for the pre-treatment group at week 3. The PTS for the post-treatment group at week 3 was similar to the pre-treatment group at 1 and 2 kHz (0-10 dB) but was intermediate between the control and pre-treatment groups at 4 and 8 kHz (23 dB). Animals pre-treated with antioxidant had a significant reduction in hair cell loss but those post-treated with antioxidant had no protection from hair cell loss. These findings demonstrate the feasibility of reduction of noise-induced hearing loss using clinically available antioxidant compounds.


Heterotopic bone formation around vessels: pilot study of a new animal model.
To achieve an easily established, safe, and reproducible animal model for the study of heterotopic bone formation around vessels, a small animal series using New Zealand White rabbits was performed. Three different dosages of recombinant human bone morphogenic protein (rhBMP-2) carried by fibrin matrix were tested. A guided tissue regeneration (GTR) membrane sheet was formed into a tube and allowed to harden; it served both to maintain the space around the vessel bundle and to separate the fibrin matrix with rhBMP-2 from skeletal muscle. Wrapped around the femoral vessel bundle and fixed in place, the tube was filled with the fibrin matrix containing rhBMP-2. The surgical site was closed in layers, and the postoperative healing was uneventful. All animals resumed their full preoperative daily activities 3-4 days after the operation. No adverse events such as wound dehiscence or infection occurred, and all animals could be sacrified at the scheduled date. Micro-computed tomography and histological investigations showed heterotopic bone formation around the vessel bundle in the medium- and high-dosage rhBMP-2 groups. An easy, safe, and reproducible animal model that allows the study of heterotopic bone formation around vessels was successfully established.


An application of image processing techniques in computed tomography image analysis.
An estimate of the thickness of subcutaneous adipose tissue at differing positions around the body was required in a study examining body composition. To eliminate human error associated with the manual placement of markers for measurements and to facilitate the collection of data from a large number of animals and image slices, automation of the process was desirable. The open-source and free image analysis program ImageJ was used. A macro procedure was created that provided the required functionality. The macro performs a number of basic image processing procedures. These include an initial process designed to remove the scanning table from the image and to center the animal in the image. This is followed by placement of a vertical line segment from the mid point of the upper border of the image to the image center. Measurements are made between automatically detected outer and inner boundaries of subcutaneous adipose tissue along this line segment. This process was repeated as the image was rotated (with the line position remaining unchanged) so that measurements around the complete circumference were obtained. Additionally, an image was created showing all detected boundary points so that sites of measurement could be checked by visual inspection. This paper provides a detailed account of this macro, illustrates the effect of its components on the image, and the applicability of standard image processing techniques to veterinary imaging.


Chaga mushroom (Inonotus obliquus) induces G0/G1 arrest and apoptosis in human hepatoma HepG2 cells.
To investigate the anti-proliferative and apoptotic effects of Chaga mushroom (Inonotus obliquus) water extract on human hepatoma cell lines, HepG2 and Hep3B cells. The cytotoxicity of Chaga extract was screened by 3-[4,5-dimethylthiazol-2-yl]-2, 5-diphenyltetrazolium bromide (MTT) assay. Morphological observation, flow cytometry analysis, Western blot were employed to elucidate the cytotoxic mechanism of Chaga extract. HepG2 cells were more sensitive to Chaga extract than Hep3B cells, as demonstrated by markedly reduced cell viability. Chaga extract inhibited the cell growth in a dose-dependent manner, which was accompanied with G0/G1-phase arrest and apoptotic cell death. In addition, G0/G1 arrest in the cell cycle was closely associated with down-regulation of p53, pRb, p27, cyclins D1, D2, E, cyclin-dependent kinase (Cdk) 2, Cdk4, and Cdk6 expression. Chaga mushroom may provide a new therapeutic option, as a potential anticancer agent, in the treatment of hepatoma.


A long-term experimental case study of the ecological effectiveness and cost effectiveness of invasive plant management in achieving conservation goals: bitou bush control in booderee national park in eastern australia.
Invasive plant management is often justified in terms of conservation goals, yet progress is rarely assessed against these broader goals, instead focussing on short-term reductions of the invader as a measure of success. Key questions commonly remain unanswered including whether invader removal reverses invader impacts and whether management itself has negative ecosystem impacts. We addressed these knowledge gaps using a seven year experimental investigation of Bitou Bush, Chrysanthemoides monilifera subsp. rotundata. Our case study took advantage of the realities of applied management interventions for Bitou Bush to assess whether it is a driver or passenger of environmental change, and quantified conservation benefits relative to management costs of different treatment regimes. Among treatments examined, spraying with herbicide followed by burning and subsequent re-spraying (spray-fire-spray) proved the most effective for reducing the number of individuals and cover of Bitou Bush. Other treatment regimes (e.g. fire followed by spraying, or two fires in succession) were less effective or even exacerbated Bitou Bush invasion. The spray-fire-spray regime did not increase susceptibility of treated areas to re-invasion by Bitou Bush or other exotic species. This regime significantly reduced plant species richness and cover, but these effects were short-lived. The spray-fire-spray regime was the most cost-effective approach to controlling a highly invasive species and facilitating restoration of native plant species richness to levels characteristic of uninvaded sites. We provide a decision tree to guide management, where recommended actions depend on the outcome of post-treatment monitoring and performance against objectives. Critical to success is avoiding partial treatments and treatment sequences that may exacerbate invasive species impacts. We also show the value of taking advantage of unplanned events, such as wildfires, to achieve management objectives at reduced cost.


Comprehensive methylome characterization of Mycoplasma genitalium and Mycoplasma pneumoniae at single-base resolution.
In the bacterial world, methylation is most commonly associated with restriction-modification systems that provide a defense mechanism against invading foreign genomes. In addition, it is known that methylation plays functionally important roles, including timing of DNA replication, chromosome partitioning, DNA repair, and regulation of gene expression. However, full DNA methylome analyses are scarce due to a lack of a simple methodology for rapid and sensitive detection of common epigenetic marks (ie N(6)-methyladenine (6 mA) and N(4)-methylcytosine (4 mC)), in these organisms. Here, we use Single-Molecule Real-Time (SMRT) sequencing to determine the methylomes of two related human pathogen species, Mycoplasma genitalium G-37 and Mycoplasma pneumoniae M129, with single-base resolution. Our analysis identified two new methylation motifs not previously described in bacteria: a widespread 6 mA methylation motif common to both bacteria (5'-CTAT-3'), as well as a more complex Type I m6A sequence motif in M. pneumoniae (5'-GAN(7)TAY-3'/3'-CTN(7)ATR-5'). We identify the methyltransferase responsible for the common motif and suggest the one involved in M. pneumoniae only. Analysis of the distribution of methylation sites across the genome of M. pneumoniae suggests a potential role for methylation in regulating the cell cycle, as well as in regulation of gene expression. To our knowledge, this is one of the first direct methylome profiling studies with single-base resolution from a bacterial organism.


The female prostate revisited: perineal ultrasound and biochemical studies of female ejaculate.
Many speculations have been made on the possible existence of a "female prostate gland" and "female ejaculation." Despite several reports on the subject, controversy still exists around the "female prostate" and whether such a gland might be the source of fluid emitted during orgasm (ejaculation). To investigate the ultrasonographic, biochemical, and endoscopic features in two women who reported actual ejaculations during orgasm. Perineal ultrasound studies, as well as biochemical characteristics of ejaculate and urethroscopy, have been performed in two women. Two premenopausal women--44 and 45 years of age--who actually reported fluid expulsion (ejaculation) during orgasm have been investigated. Ultrasound imaging, biochemical studies of the ejaculated fluid, and endoscopy of the urethra have been used to identify a prostate in the female. Ejaculated fluid parameters have been compared to voided urine samples. On high-definition perineal ultrasound images, a structure was identified consistent with the gland tissue surrounding the entire length of the female urethra. On urethroscopy, one midline opening (duct) was seen just inside the external meatus in the six-o'clock position. Biochemically, the fluid emitted during orgasm showed all the parameters found in prostate plasma in contrast to the values measured in voided urine. Data of the two women presented further underline the concept of the female prostate both as an organ itself and as the source of female ejaculation.


Demonstration of a specific receptor for human C5a anaphylatoxin on murine macrophages.
Human C5a anaphylatoxin is known to be a potent mediator of the acute inflammatory response. It serves to trigger a wide variety of neutrophil responses after binding to a specific cellular receptor. We have now demonstrated that this bioactive glycopolypeptide is also bound to a specific receptor found on murine resident peritoneal macrophages, thioglycollate-induced exudate macrophages, and the murine cell line P388D1. The apparent Kd of the C5a receptors. Resident macrophages appear to have an average of 2 x 10(5) binding sites per cell, whereas thioglycollate- induced cells have only 4-5 x 10(4) binding sites. The continuous cell line P388D1 is intermediate between these two cell types, exhibiting 8-10 x 10(4) C5a receptors per cell. Neither murine lymphocytes nor the parent cell line P388 displays a measurable number of C5a receptors. Macrophage receptor-C5a binding interactions are followed by cellular uptake and degradation of 125I-C5a, much as is observed with neutrophils. As demonstrated in another paper, binding of C5a to macrophages results in augmentation of the primary humoral immune response as well as enhancement of mixed lymphocyte reactions. These observations suggest that C5a should not only be considered as an acute inflammatory mediator but as an immunopotentiating modulator as well, thus serving as a critical link between complement activation and subsequent immune responses.


2P2I HUNTER: a tool for filtering orthosteric protein-protein interaction modulators via a dedicated support vector machine.
Over the last 10 years, protein-protein interactions (PPIs) have shown increasing potential as new therapeutic targets. As a consequence, PPIs are today the most screened target class in high-throughput screening (HTS). The development of broad chemical libraries dedicated to these particular targets is essential; however, the chemical space associated with this 'high-hanging fruit' is still under debate. Here, we analyse the properties of 40 non-redundant small molecules present in the 2P2I database (http://2p2idb.cnrs-mrs.fr/) to define a general profile of orthosteric inhibitors and propose an original protocol to filter general screening libraries using a support vector machine (SVM) with 11 standard Dragon molecular descriptors. The filtering protocol has been validated using external datasets from PubChem BioAssay and results from in-house screening campaigns. This external blind validation demonstrated the ability of the SVM model to reduce the size of the filtered chemical library by eliminating up to 96% of the compounds as well as enhancing the proportion of active compounds by up to a factor of 8. We believe that the resulting chemical space identified in this paper will provide the scientific community with a concrete support to search for PPI inhibitors during HTS campaigns.


Outcome after open reduction and internal fixation of capitellar and trochlear fractures.
Capitellar and trochlear fractures are uncommon fractures of the distal aspect of the humerus. There is limited information about the functional outcome of patients managed with open reduction and internal fixation. The functional outcome of twenty-eight patients, with a mean age (and standard deviation) of 43 +/- 13 years, who were treated with open reduction and internal fixation for capitellar and trochlear fractures was evaluated at a mean duration of follow-up of 56 +/- 33 months. Patient outcomes were assessed with physical and radiographic examination, range-of-motion measurements, strength testing, and self-reported questionnaires (Short Form-36, Mayo Elbow Performance Index, American Shoulder and Elbow Surgeons Elbow Assessment Form, and Patient-Rated Elbow Evaluation scales). Eleven fractures involved the capitellum with or without fracture of the lateral ridge of the trochlea, four involved the capitellum and trochlea as one piece, and thirteen involved the capitellum and trochlea as separate fragments. These fractures were further characterized by the presence or absence of posterior comminution. Fourteen patients had isolated fractures, and fourteen had other elbow, forearm, or wrist injuries. Patients with more complex fractures required more extensive surgery, had more complications resulting in secondary procedures, and had poorer outcomes compared with those with simple fractures. The average score on the Mayo Elbow Performance Index (91 +/- 11), the average quality-of-life scores (46 on the physical component and 50 on the mental component of the Short Form-36), and the average range of motion (19 degrees to 138 degrees ) suggest favorable patient outcomes overall. Two comminuted fractures did not unite and required conversion to a total elbow arthroplasty. Patients with isolated noncomminuted capitellar and/or trochlear fractures have better results than those with more complex fractures. A classification system based on the radiographic patterns of these fractures is recommended.


Reproductive Alterations in Chronically Exposed Female Mice to Environmentally Relevant Doses of a Mixture of Phthalates and Alkylphenols.
Endocrine-disrupting chemicals (EDCs) are exogenous compounds that modify hormone biosynthesis, causing adverse effects to human health. Among them, phthalates and alkylphenols are important due to their wide use in plastics, detergents, personal care products, cosmetics, and food packaging. However, their conjoint effects over reproductive female health have not been addressed. The aim of this work was to test the effect of chronically exposed female mice to a mixture of three phthalates [bis (2-ethylhexyl), dibutyl, and benzyl butyl] and two alkylphenols (4-nonylphenol and 4-tert-octylphenol) from conception to adulthood at environmentally relevant doses. These EDCs were administered in two doses: one below the minimal risk dose to cause adverse effects on human development and reproduction [1 mg/kg body weight (BW)/d of the total mixture] and the other one based on the reference value close to occupational exposure in humans (10 mg/kg BW/d of the total mixture). Our results show that both doses had similar effects regarding the uterus and ovary relative weight, estrous cyclicity, serum levels of progesterone and 17β-estradiol, and expression of key elements in the steroidogenesis pathway (acute steroidogenic regulatory protein and CYP19A1). However, only the 1-mg/kg BW/d dose delayed the onset of puberty and the transition from preantral to antral follicles, whereas the 10-mg/kg BW/d dose decreased the number of antral follicles and gonadotropin receptor expression. In addition, we observed changes in several fertility parameters in exposed females and in their progeny (F2 generation). In conclusion, our results indicate that chronic exposure to a complex EDC mixture, at environmentally relevant doses, modifies reproductive parameters in female mice.


Brain protection using autologous bone marrow cell, metalloproteinase inhibitors, and metabolic treatment in cerebral ischemia.
Despite advances in imaging, understanding the underlying pathways, and clinical translation of animal models of disease there remains an urgent need for therapies that reduce brain damage after stroke and promote functional recovery in patients. Blocking oxidant radicals, reducing matrix metalloproteinase-induced neuronal damage, and use of stem cell therapy have been proposed and tested individually in prior studies. Here we provide a comprehensive integrative management approach to reducing damage and promoting recovery by combining biological therapies targeting these areas. In a rat model of transient cerebral ischemia (middle cerebral artery occlusion) gene delivery vectors were used to overexpress tissue inhibitor of matrix metalloproteinase 1 and 2 (TIMP1 and TIMP2) 3 days before ischemia. After occlusion, autologous bone marrow cells alone or in combination with agents to improve NO bioavailability were administered intraarterially. When infarct size, BrdU incorporation, and motor function recovery were determined in the treatment groups the largest beneficial effect was seen in rats receiving the triple combined therapy, surpassing effects of single or double therapies. Our study highlights the utility of combined drug, gene, and cell therapy in the treatment of stroke.


Rapid Recovery From Cortical Blindness Caused by an Old Cerebral Infarction.
When the primary visual cortex (V1) is damaged, cortical blindness results. However, visual information obtained from the superior colliculus (SC) or direct thalamic afferents to higher visual cortices produces unconscious visual functions called blindsight. Alarming visual stimuli suggesting the approach of a predator are known to trigger escape behaviors via visual information mediated by the SC and amygdala in small animals, and salient and dynamic visual stimuli also produce some conscious visual experience even in patients with blindsight. Fresh cortical blindness sometimes recovers spontaneously in patients with fresh cerebral damages, and recovery can be accelerated by early rehabilitation. However, the mechanisms underlying recovery are not well-known. We analyzed a patient with cortical blindness caused by an old cerebral infarction. After repeated presentation of alarming visual stimuli, the ability to detect visual stimuli in the impaired visual field showed behavioral short-term improvement (STI) within a few minutes. Repeated behavioral STI induction was followed by behavioral long-term improvement (LTI) lasting more than several days. After behavioral LTI, the patient partially recovered the ability to read letters presented in the impaired visual field. The behavioral STI experiment, which can be performed within 10 min, may serve as a clinical screening test for anticipating recovery from cortical blindness.


Characterization of four rice UEV1 genes required for Lys63-linked polyubiquitination and distinct functions.
The error-free branch of the DNA-damage tolerance (DDT) pathway is orchestrated by Lys63-linked polyubiquitination of proliferating cell nuclear antigen (PCNA), and this polyubiquitination is mediated by a Ubc13-Uev complex in yeast. We have previously cloned OsUBC13 from rice, whose product functions as an E2 to promote Lys63-linked ubiquitin chain assembly in the presence of yeast or human Uev. Here we identify four highly conserved UEV1 genes in rice whose products are able to form stable heterodimers with OsUbc13 and mediate Lys63-linked ubiquitin chain assembly. Expression of OsUEV1s is able to rescue the yeast mms2 mutant from death caused by DNA-damaging agents. Interestingly, OsUev1A contains a unique C-terminal tail with a conserved prenylation site not found in the other three OsUev1s, and this post-translational modification appears to be required for its unique subcellular distribution and association with the membrane. The analysis of OsUEV1 expression profiles obtained from the Genevestigator database indicates that these genes are differentially regulated. We speculate that different OsUev1s play distinct roles by serving as a regulatory subunit of the Ubc13-Uev1 complex to respond to diverse cellular, developmental and environmental signals.


Morphine, hypotension, and adverse outcomes among preterm neonates: who's to blame? Secondary results from the NEOPAIN trial.
Hypotension occurs commonly among preterm neonates, but its cause and consequences remain unclear. Secondary data analyses from the NEOPAIN trial identified the clinical factors associated with hypotension and examined the contributions of morphine treatment or hypotension to severe intraventricular hemorrhage (IVH) (grades 3 and 4), any IVH (grades 1-4), or death. In the NEOPAIN trial, 898 ventilated neonates between 23 and 32 weeks of gestation were enrolled, with equal numbers randomized to receive masked morphine or placebo infusions. Additional doses of open-label morphine were administered as necessary by medical staff members. IVH was diagnosed with centralized readings of early and late cranial ultrasonograms. Hypotension was assessed before study drug infusion, during the loading dose, and at 24 and 72 hours during study drug infusion. Logistic regression analyses with stepdown elimination identified the predictor factors associated with the hypotension, severe IVH, any IVH, or death outcomes at each time point. Hypotension was associated with 23 to 26 weeks of gestation, morphine infusions, severity of illness, additional morphine doses, and prior hypotension. Severe IVH was associated with shorter gestation, higher Clinical Risk Index for Babies scores, no prenatal steroids, pulmonary hemorrhage, hypotension before the loading dose, and morphine doses before intubation and at 25 to 72 hours. Neonatal deaths were associated with 23 to 26 weeks of gestation, higher Clinical Risk Index for Babies scores, pulmonary hemorrhage, patent ductus arteriosus, thrombocytopenia, and hypotension before the loading dose. Morphine infusions were not a significant factor in logistic models for severe IVH, any IVH, or death. Preemptive morphine infusions, additional morphine, and lower gestational age were associated with hypotension among preterm neonates. Severe IVH, any IVH, and death were associated with preexisting hypotension, but morphine therapy did not contribute to these outcomes. Morphine infusions, although they cause hypotension, can be used safely for most preterm neonates but should be used cautiously for 23- to 26-week neonates and those with preexisting hypotension.


Polarized gamma-ray emission from the crab.
Pulsar systems accelerate particles to immense energies. The detailed functioning of these engines is still poorly understood, but polarization measurements of high-energy radiation may allow us to locate where the particles are accelerated. We have detected polarized gamma rays from the vicinity of the Crab pulsar using data from the spectrometer on the International Gamma-Ray Astrophysics Laboratory satellite. Our results show polarization with an electric vector aligned with the spin axis of the neutron star, demonstrating that a substantial fraction of the high-energy electrons responsible for the polarized photons are produced in a highly ordered structure close to the pulsar.


Management of fluoride induced testicular disorders by calcium and vitamin-E co-administration in the albino rat.
Fluoride contamination of drinking water can disrupt male gametogenesis and steroidogenesis and induce testicular oxidative stress. Treatment of rats with sodium fluoride at the dose of 20 mg/kg/day for 28 days resulted in significant diminution of testicular Delta5,3beta-hydroxysteroid dehydrogenase (HSD) and 17beta-hydroxysteroid dehydrogenase (HSD) activities and low plasma levels of testosterone, follicular stimulating hormone (FSH) and leutinizing hormone (LH). Spermatogenesis inhibited after sodium fluoride treatment has been assessed here by the quantification of different generation of germ cells like spermatogonia A (ASg), preleptotene spermatocyte (PLSc), midpachytene spermatocyte (MPSc) and step 7 spermatid (7Sd) at stage VII of seminiferous epithelial cycle. Furthermore, fluoride treatment was associated with low activities of testicular, prostatic and epididymal catalase (CAT), superoxide dismutase (SOD) and peroxidase along with elevation of malondialdehyde (MDA) and conjugated dienes (CD) in those tissues. Co-administration of calcium and Vitamin-E with fluoride resulted in a significant recovery from testicular disorders and oxidative stress in the testis and male accessory sex organs. The results of this study demonstrate that fluoride exposure, at the dose available in drinking water in contaminated areas, led to inhibition of testicular gametogenesis and steroidogenesis in association with oxidative stress in the testis and male accessory sex organs, which are protected significantly by dietary agents like Vitamin-E and calcium.


Autoproteolysis in hedgehog protein biogenesis.
Extracellular signaling proteins encoded by the hedgehog (hh) multigene family are responsible for the patterning of a variety of embryonic structures in vertebrates and invertebrates. The Drosophila hh gene has now been shown to generate two predominant protein species that are derived by an internal autoproteolytic cleavage of a larger precursor. Mutations that reduced the efficiency of autoproteolysis in vitro diminished precursor cleavage in vivo and also impaired the signaling and patterning activities of the HH protein. The two HH protein species exhibited distinctive biochemical properties and tissue distribution, and these differences suggest a mechanism that could account for the long- and short-range signaling activities of HH in vivo.


Association Between Appendectomy and Clostridium difficile Infection.
Recent theory proposes that the appendix functions as a reservoir for commensal bacteria, and serves to re-inoculate the colon with normal flora in the event of pathogen exposure or purging of intestinal flora. If true, we reasoned that flora from a normal appendix could provide protection against Clostridium difficile. We conducted this investigation to examine the protective effect of an intact appendix and test the hypothesis that prior appendectomy will be more common among patients with a positive test for C. difficile as compared with patients who test negative. We contacted patients who had undergone C. difficile testing and asked them whether or not they had a prior appendectomy. Using their responses and results from Toxin A & B EIA tests, we calculated the difference in appendectomy rates between those who tested positive for C. difficile, and those who tested negative. We considered a positive 15% absolute difference to represent a significant increase in appendectomy rate. We enrolled 257 patients. Among the 136 who tested positive for C. difficile, 27 (19.9%) had prior appendectomies, while among 121 patients testing negative for C. difficile, 38 (31.4%) had prior appendectomies, yielding a difference in appendectomy rates of -11.6% (95% Confidence Interval: -21.6% to -0.9%). The rate of prior appendectomy was actually lower among patients with a positive C. difficile test as compared to those with a negative test. Conversely, patients who tested positive for C. difficile were more likely to have an intact appendix than those who tested negative. These results suggest that rather than being protective, an intact appendix appears to promote C. difficile acquisition, carriage, and disease. Clostridium difficile; Appendix; Appendectomy; Microbial reservoir; Infection.


Subclinical cognitive decline in middle-age is associated with reduced task-induced deactivation of the brain's default mode network.
Cognitive abilities decline with age, but with considerable individual variation. The neurobiological correlate of this variation is not well described. Functional brain imaging studies have demonstrated reduced task-induced deactivation (TID) of the brain's default mode network (DMN) in a wide range of neurodegenerative diseases involving cognitive symptoms, in conditions with increased risk of Alzheimer's disease, and even in advanced but healthy aging. Here, we investigated brain activation and deactivation during a visual-motor task in 185 clinically healthy males from a Danish birth cohort, whose cognitive function was assessed in youth and midlife. Using each individual as his own control, we defined a group with a large degree of cognitive decline, and a control group. When correcting for effects of total cerebral blood flow and hemoglobin level, we found reduced TID in the posterior region of the DMN in the cognitive decline group compared to the control group. Furthermore, increased visual activation response was found in the cognitive decline group, indicating that the TID reduction was not exclusively due to overall impaired vascular reactivity. These results suggest a neurobiological basis for subclinical cognitive decline in late midlife, which includes TID alterations similar to the pattern seen in patients with AD and mild cognitive impairment. Hence, TID reduction might be suggested as an early marker for subtle cognitive decline in aging.


Light-scattering properties of plate and column ice crystals generated in a laboratory cold chamber.
Angular scattering properties of ice crystal particles generated in a laboratory cloud chamber are measured with a lightweight polar nephelometer with a diode laser beam. This cloud chamber produces distinct plate and hollow column ice crystal types for light-scattering experiments and provides a controlled test bed for comparison with results computed from theory. Ice clouds composed predominantly of plates and hollow columns generated noticeable 22 degrees and 46 degrees halo patterns, which are predicted from geometric ray-tracing calculations. With the measured ice crystal shape and size distribution, the angular scattering patterns computed from geometrical optics with a significant contribution by rough surfaces closely match those observed from the nephelometer.


A multilectin affinity approach for comparative glycoprotein profiling of rheumatoid arthritis and spondyloarthropathy.
Arthritis refers to inflammation of joints and includes common disorders such as rheumatoid arthritis (RA) and spondyloarthropathies (SpAs). These diseases differ mainly in terms of their clinical manifestations and the underlying pathogenesis. Glycoproteins in synovial fluid might reflect the disease activity status in the joints affected by arthritis; yet they have not been systematically studied previously. Although markers have been described for assisting in the diagnosis of RA, there are currently no known biomarkers for SpA. We sought to determine the relative abundance of glycoproteins in RA and SpA by lectin affinity chromatography coupled to iTRAQ labeling and LC-MS/MS analysis. We also used ELISA to validate the overexpression of VCAM-1, one of the candidate proteins identified in this study, in synovial fluid from RA patients. We identified proteins that were previously reported to be overexpressed in RA including metalloproteinase inhibitor 1 (TIMP1), myeloperoxidase (MPO) and several S100 proteins. In addition, we discovered several novel candidates that were overexpressed in SpA including Apolipoproteins C-II and C-III and the SUN domain-containing protein 3 (SUN3). Novel molecules found overexpressed in RA included extracellular matrix protein 1 (ECM1) and lumican (LUM). We validated one of the candidate biomarkers, vascular cell adhesion molecule 1 (VCAM1), in 20 RA and SpA samples using ELISA and confirmed its overexpression in RA (p-value <0.01). Our quantitative glycoproteomic approach to study arthritic disorders should open up new avenues for additional proteomics-based discovery studies in rheumatological disorders.


Structure and function of immunoglobulins.
Immunoglobulins are heterodimeric proteins composed of 2 heavy and 2 light chains. They can be separated functionally into variable domains that bind antigens and constant domains that specify effector functions, such as activation of complement or binding to Fc receptors. The variable domains are created by means of a complex series of gene rearrangement events and can then be subjected to somatic hypermutation after exposure to antigen to allow affinity maturation. Each variable domain can be split into 3 regions of sequence variability termed the complementarity-determining regions (CDRs) and 4 regions of relatively constant sequence termed the framework regions. The 3 CDRs of the heavy chain are paired with the 3 CDRs of the light chain to form the antigen-binding site, as classically defined. The constant domains of the heavy chain can be switched to allow altered effector function while maintaining antigen specificity. There are 5 main classes of heavy chain constant domains. Each class defines the IgM, IgG, IgA, IgD, and IgE isotypes. IgG can be split into 4 subclasses, IgG1, IgG2, IgG3, and IgG4, each with its own biologic properties, and IgA can similarly be split into IgA1 and IgA2.


Serum Biomarkers for Prediction of Response to Methotrexate Monotherapy in Early Rheumatoid Arthritis: Results from the SWEFOT Trial.
To investigate baseline levels of 12 serum biomarkers that constitute a multibiomarker disease activity test, as predictors of response to methotrexate (MTX) in patients with early rheumatoid arthritis (eRA). In 298 patients from the Swedish Pharmacotherapy (SWEFOT) clinical trial, baseline serum levels of 12 proteins were analyzed for association with disease activity based on the 28-joint count Disease Activity Score (DAS28) after 3 months of MTX monotherapy using uni-/multivariate logistic regression. Primary outcome was low disease activity (LDA; DAS28 ≤ 3.2). Of 298 patients, 104 achieved LDA after 3 months on MTX. Four of the 12 biomarkers [C-reactive protein (CRP), leptin, tumor necrosis factor receptor I (TNF-RI), and vascular cell adhesion molecule 1 (VCAM-1)] significantly predicted LDA based on stepwise logistic regression analysis. Dichotomization of patients using receiver-operating characteristic curve analysis-based cutoffs for these biomarkers showed significantly higher proportions with LDA among patients with lower versus higher levels of CRP or leptin (40% vs 23%, p = 0.004, and 40% vs 25%, p = 0.011, respectively), as well as among those with higher versus lower levels of TNF-RI or VCAM-1 (43% vs 27%, p = 0.004, and 41% vs 25%, p = 0.004, respectively). Combined score based on these biomarkers, adjusted for known predictors of LDA (smoking, sex, and age), associated with decreased chance of LDA (adjusted OR 0.45, 95% CI 0.32-0.62). Low baseline levels of CRP and leptin, and high baseline levels of TNF-RI and VCAM-1 were associated with LDA after 3 months of MTX therapy in patients with eRA. Combination of these 4 biomarkers increased accuracy of prediction. [Trial registration number: NCT00764725].


Post-disaster mental health problems and the utilization of mental health services: a four-year longitudinal comparative study.
This study examined mental health problems and mental health services (MHS) utilization after a fireworks disaster among adult survivors and a comparison group. The disaster took place on May 13, 2000, in the city of Enschede, The Netherlands. Victims (N=662) participated in a survey 2-3 weeks (T1), 18 months (T2) and 4 years (T3) post-disaster. The comparison group consisted of non-affected people from another city (N=526). They participated at T2 and T3. Victims used MHS more often than the comparison group in the 12-month period before T2 and T3 (OR 3.9 and 2.4). Victims with severe depression and anxiety symptoms at T2 used MHS more often than participants in the comparison group with these symptoms (OR 2.6 and 2.0). After 4 years, MHS utilization among participants in both groups with anxiety symptoms did not differ, suggesting attenuation of the observed effects. Results suggest that after a disaster survivors with mental health problems are less reluctant to use MHS than under normal circumstances.


Road user behaviour, attitudes and crashes: a survey of Aboriginal and Torres Strait Islander people in Australia.
This study aimed to describe road user behaviour, attitudes and crashes in Aboriginal and Torres Strait Islander people in four urban, regional and remote communities located in New South Wales (NSW) and South Australia (SA). Face-to-face surveys were administered to clients (n=625) in Aboriginal Community Controlled Health Services (ACCHS). All Aboriginal and Torres Strait Islander clients attending the ACCHS for any reason were approached to participate over a 2-week period. Surveys included questions on sociodemographic factors, crash involvement, road behaviours and road safety attitudes drawn from tools used in national surveys. The participation rate was high (69%-75%). Seat belt wearing rates were very high, particularly in the front of a car, although rear seat belt wearing rates in SA (77%) were substantially lower than in NSW (93%). Among drivers, 11% reported always or mostly driving 10 km/hour over the speed limit, and this was higher among drivers in SA (13.4%). Drivers aged 55 years and over and/or women were more likely to report that they do not drink at any time or restricted what they drank when driving. These results enable comparison with the Community Attitude to Road Safety survey conducted Australia-wide in 2013. This study confirms that Aboriginal and Torres Strait Islander people are inclined to report attitudes or road safety behaviours similar to the rest of the population; however, rear restraint use was lower and self-reported speeding was higher. These issues are likely attributable to transport options and geography in remote communities, which can contribute to overcrowding and unsafe driving practices.


Entacapone prolongs levodopa response in a one month double blind study in parkinsonian patients with levodopa related fluctuations.
To establish, in a double blind manner, the antiparkinsonian effects of repeated dosing with entacapone, a peripheral COMT inhibitor. A one month, cross over study was conducted. During the two four-week treatment periods, entacapone (200 mg) or placebo was given with each levodopa dose four to 10 times daily. Motor responses were repeatedly quantified using the motor part of UPDRS. Plasma levodopa and its metabolites were measured. Entacapone prolonged the availability of levodopa in the plasma and thus to the brain by decreasing its peripheral O-methylation and slowing its elimination rate, without affecting the maximum plasma levodopa concentration or the time to maximum concentration. Corresponding with the pharmacokinetic findings, entacapone prolonged the duration of motor response to an individual levodopa/DDC inhibitor dose by 34 minutes (24%, P = 0.001) and dyskinesiae by 39 minutes (37%, P = 0.002) compared with placebo, without affecting their magnitude or starting time. Entacapone treatment resulted in a reduction of 16% in the mean total daily levodopa dose due to dyskinesiae. Also, according to the home diaries, the mean daily "on" time increased by 2.1 hours compared with placebo, despite the lowered mean levodopa intake. The efficacy of repeated entacapone dosing as an adjuvant to levodopa/DDC inhibitor treatment for Parkinson's disease with levodopa related fluctuations is verified.


Socioeconomic outcomes of combined spine surgery and functional restoration in workers' compensation spinal disorders with matched controls.
A longitudinal cohort study (n = 448) comparing functionally restored discectomy (n = 123) and fusion (n = 101) workers' compensation patients to matched, unoperated control patients (n = 123 and n = 101, respectively). To determine successful treatment outcomes uniquely important in a workers' compensation environment when spine surgery is combined with comprehensive tertiary rehabilitation, to optimize anatomic and social sequelae. Multiple recent studies confirm suboptimal socioeconomic outcomes for spinal surgery for degenerative conditions in a workers' compensation venue. In other musculoskeletal regions, there is a clear relationship between the quality of postsurgical rehabilitation and the impact on disability, recurrent injury, and future health care use. It is hypothesized that poor surgical outcomes in compensation injuries may result from outmoded postoperative methods, rather than failures of patient selection or surgical technique. No previous combination of surgery plus rehabilitation has been carefully evaluated with disabled workers undergoing spine surgery. Functional restoration is an individualized medically directed, interdisciplinary program using quantitatively directed exercise progression, psychotherapeutic interventions, and monitoring of specific socioeconomic outcomes for chronically disabled workers. This study prospectively evaluated a cohort of consecutive functional restoration program graduates (n = 1202). Two surgical groups, discectomy (n = 123) and fusion (n = 101) were matched to two groups of unoperated control patients, control/discectomy and control/fusion, selected from the same cohort of patients with chronic spinal disorders based on age, gender, race, length of disability, and workers' compensation jurisdiction. A structured clinical interview was administered 12 months after program completion, with a contact rate of 95% to 98%. Socioeconomic outcomes for work return, health care use, and recurrent lost-time injury were assessed. All groups demonstrated a return-to-work incidence of more than 85%, but work retention at 1 year was higher for the fusion group than for the discectomy or control/fusion groups. Health care use was significantly higher for the discectomy group than the control/discectomy or fusion groups for reoperation (8% vs. 4%/ 2%), as well as other factors. All groups showed comparable recurrent lost-time injury rates (2-3.3%), and made comparable improvements in prospectively collected physical and psychological measures. Discectomy patients had work, health care utilization, and recurrent injury outcomes comparable with those for unoperated control patients. Fusion patients had better outcomes of work retention, reoperation, and health care use compared with the unoperated control patients and even with discectomy patients, in spite of more cases of previous surgery and greater duration of disability. The discectomy and fusion cohorts of operated chronic spinal disorder compensation patients with subsequent functional restoration had the best documented outcomes found in the literature for this population. In spite of the common presumption that spine surgery patients fare poorly in a workers' compensation environment, these results demonstrate that such patients can show remarkably successful objective outcomes if accompanied by effective rehabilitation, documenting efficacy and clinical utility. A new clinical approach is required to evaluate prospectively the combination of surgery and rehabilitation in chronic pain/disability workers' compensation patients, in which the surgical role is to correct an anatomic lesion, but the socioeconomic outcomes either occur spontaneously or are effected through some form of rehabilitation.


AKT promotes rRNA synthesis and cooperates with c-MYC to stimulate ribosome biogenesis in cancer.
Precise regulation of ribosome biogenesis is fundamental to maintain normal cell growth and proliferation, and accelerated ribosome biogenesis is associated with malignant transformation. Here, we show that the kinase AKT regulates ribosome biogenesis at multiple levels to promote ribosomal RNA (rRNA) synthesis. Transcription elongation by RNA polymerase I, which synthesizes rRNA, required continuous AKT-dependent signaling, an effect independent of AKT's role in activating the translation-promoting complex mTORC1 (mammalian target of rapamycin complex 1). Sustained inhibition of AKT and mTORC1 cooperated to reduce rRNA synthesis and ribosome biogenesis by additionally limiting RNA polymerase I loading and pre-rRNA processing. In the absence of growth factors, constitutively active AKT increased synthesis of rRNA, ribosome biogenesis, and cell growth. Furthermore, AKT cooperated with the transcription factor c-MYC to synergistically activate rRNA synthesis and ribosome biogenesis, defining a network involving AKT, mTORC1, and c-MYC as a master controller of cell growth. Maximal activation of c-MYC-dependent rRNA synthesis in lymphoma cells required AKT activity. Moreover, inhibition of AKT-dependent rRNA transcription was associated with increased lymphoma cell death by apoptosis. These data indicate that decreased ribosome biogenesis is likely to be a fundamental component of the therapeutic response to AKT inhibitors in cancer.


Screening a combinatorial peptide library to develop a human glandular kallikrein 2-activated prodrug as targeted therapy for prostate cancer.
Prostate cancer cells secrete the unique protease human glandular kallikrein 2 (hK2) that represents a target for proteolytic activation of cytotoxic prodrugs. The objective of this study was to identify hK2-selective peptide substrates that could be coupled to a cytotoxic analogue of thapsigargin, a potent inhibitor of the sarcoplasmic/endoplasmic reticulum calcium ATPase pump that induces cell proliferation-independent apoptosis through dysregulation of intracellular calcium levels. To identify peptide sequence requirements for hK2, a combination of membrane-bound peptides (SPOT analysis) and combinatorial chemistry using fluorescence-quenched peptide substrates was used. Peptide substrates were then coupled to 8-O-(12[L-leucinoylamino]dodecanoyl)-8-O-debutanoylthapsigargin (L12ADT), a potent analogue of thapsigargin, to produce a prodrug that was then characterized for hK2 hydrolysis, plasma stability, and in vitro cytotoxicity. Both techniques indicated that a peptide with two arginines NH2-terminal of the scissile bond produced the highest rates of hydrolysis. A lead peptide substrate with the sequence Gly-Lys-Ala-Phe-Arg-Arg (GKAFRR) was hydrolyzed by hK2 with a Km of 26.5 micromol/L, kcat of 1.09 s(-1), and a kcat/Km ratio of 41,132 s(-1) mol/L(-1). The GKAFRR-L12ADT prodrug was rapidly hydrolyzed by hK2 and was stable in plasma, whereas the GKAFRR-L peptide substrate was unstable in human plasma. The hK2-activated thapsigargin prodrug was not activated by cathepsin B, cathepsin D, and urokinase but was an excellent substrate for plasmin. The GKAFRR-L12ADT was selectively cytotoxic in vitro to cancer cells in the presence of enzymatically active hK2. The hK2-activated thapsigargin prodrug represents potential novel targeted therapy for prostate cancer.


Collection of erythroid progenitor cells by cytapheresis of peripheral blood of normal donors.
Human peripheral blood may be an alternative to bone marrow as a source of cells for hemopoietic engraftment. The ability to collect large numbers of circulating granulocyte-macrophage progenitors provides support for this contention. In the present study of cells from normal granulocyte donors, the cell concentrates obtained by cytapheresis were shown to contain even greater numbers of primitive erythroid precursors (153-956 [median 647] per 10(6) mononuclear cells) than would be predicted from the peripheral blood mononuclear cell counts. Moreover, the number of primitive erythroid precursors harvested correlated significantly with the number of granulocyte-macrophage progenitors obtained and with the total lymphocyte collection. These observations further substantiate the validity of transplanting peripheral blood as hemopoietic tissue.


Parental agreement of reporting parent to child aggression using the Conflict Tactics Scales.
This study examined mothers' and fathers' reporting congruency using the Parent-Child Conflict Tactics Scales. We asked if the mother's report of the father's parenting aggression was consistent with the father's self-report of parenting aggression and if the father's report of the mother's parenting aggression was consistent with the mother's self- report of those same behaviors. We assessed moderators of parental reporting congruency: severity of the aggression, interparental conflict, child temperament, and child gender. Participants were from the Child Development Project, a longitudinal study beginning when children were in kindergarten. The analyses herein included 163 children for whom 2 parents provided data about their own and their spouse or partner's behavior toward the child. Most parents (87%) were married. Mothers and fathers independently completed the Parent-Child Conflict Tactics Scale, both with respect to their own behavior toward the child and with respect to their partner's behavior toward the child. Mothers completed the retrospective Infant Characteristics Questionnaire to assess child temperament. Mothers and fathers completed measures of interparental conflict. Both fathers and mothers self-reported more frequently engaging in each behavior than the other parent reported they did. Parents were more congruent on items assessing harsher parenting behavior. Furthermore, there was more agreement between parents regarding fathers' behavior than mothers' behavior. Analyses of interparental conflict, child difficult temperament, and child gender as moderators yielded findings suggesting that mothers' and fathers' reports of their own and their spouses' harsh parenting behaviors were more concordant in couples with low levels of conflict, for children with easy temperaments, and for boys versus girls. Prior studies indicate only a moderate level of agreement in couples' reports of violence between intimate partners and suggest that perpetrators tend to underreport their use of aggression. The results of this study suggest that parents may be more consistent in their reports of parent to child violence using the Parent-Child Conflict Tactics Scales than they are when reporting intimate partner violence. The results suggest that parental reports of their spouse's parent to child aggression are reliable.


A bivariate measurement error model for semicontinuous and continuous variables: Application to nutritional epidemiology.
Semicontinuous data in the form of a mixture of a large portion of zero values and continuously distributed positive values frequently arise in many areas of biostatistics. This article is motivated by the analysis of relationships between disease outcomes and intakes of episodically consumed dietary components. An important aspect of studies in nutritional epidemiology is that true diet is unobservable and commonly evaluated by food frequency questionnaires with substantial measurement error. Following the regression calibration approach for measurement error correction, unknown individual intakes in the risk model are replaced by their conditional expectations given mismeasured intakes and other model covariates. Those regression calibration predictors are estimated using short-term unbiased reference measurements in a calibration substudy. Since dietary intakes are often "energy-adjusted," e.g., by using ratios of the intake of interest to total energy intake, the correct estimation of the regression calibration predictor for each energy-adjusted episodically consumed dietary component requires modeling short-term reference measurements of the component (a semicontinuous variable), and energy (a continuous variable) simultaneously in a bivariate model. In this article, we develop such a bivariate model, together with its application to regression calibration. We illustrate the new methodology using data from the NIH-AARP Diet and Health Study (Schatzkin et al., 2001, American Journal of Epidemiology 154, 1119-1125), and also evaluate its performance in a simulation study.


Biofilms: the stronghold of Legionella pneumophila.
Legionellosis is mostly caused by Legionella pneumophila and is defined as a severe respiratory illness with a case fatality rate ranging from 5% to 80%. L. pneumophila is ubiquitous in natural and anthropogenic water systems. L. pneumophila is transmitted by inhalation of contaminated aerosols produced by a variety of devices. While L. pneumophila replicates within environmental protozoa, colonization and persistence in its natural environment are also mediated by biofilm formation and colonization within multispecies microbial communities. There is now evidence that some legionellosis outbreaks are correlated with the presence of biofilms. Thus, preventing biofilm formation appears as one of the strategies to reduce water system contamination. However, we lack information about the chemical and biophysical conditions, as well as the molecular mechanisms that allow the production of biofilms by L. pneumophila. Here, we discuss the molecular basis of biofilm formation by L. pneumophila and the roles of other microbial species in L. pneumophila biofilm colonization. In addition, we discuss the protective roles of biofilms against current L. pneumophila sanitation strategies along with the initial data available on the regulation of L. pneumophila biofilm formation.


DNA triplex structure, thermodynamics, and destabilisation: insight from molecular simulations.
Molecular dynamics simulations are used to elucidate the structure and thermodynamics of DNA triplexes associated with the neurodegenerative disease Friedreich's ataxia (FRDA), as well as complexes of these triplexes with the small molecule netropsin, which is known to destabilise triplexes. The ability of molecular simulations in explicit solvent to accurately capture triplex thermodynamics is verified for the first time, with the free energy to dissociate a 15-base antiparallel purine triplex-forming oligomer (TFO) from the duplex found to be slightly higher than reported experimentally. The presence of netropsin in the minor groove destabilises the triplex as expected, reducing the dissociation free energy by approximately 50%. Netropsin binding is associated with localised narrowing of the minor groove near netropsin, an effect that has previously been under contention. This leads to localised widening of the major groove, weakening hydrogen bonds between the TFO and duplex. Consequently, destabilisation is found to be highly localised, occurring only when netropsin is bound directly opposite the TFO. The simulations also suggest that near saturation of the minor groove with ligand is required for complete triplex dissociation. A structural analysis of the DNA triplexes that can form with the FRDA-related duplex sequence indicates that the triplex with a parallel homopyrimidine TFO is likely to be more stable than the antiparallel homopurine-TFO triplex, which may have implications for disease onset and treatment.


Mannich-type addition of 1,3-dicarbonyl compounds to chiral tert-butanesulfinyltrifluoroacetaldimines. Mechanistic aspects and chiroptical studies.
Mannich-type addition of 1,3-dicarbonyl compounds to (SS)-N-tert-butanesulfinyltrifluoroacetaldimine has been carried out under dramatically different conditions. The stereochemical outcome was quite different when the reaction was carried out under solvent-free conditions, at high temperature and without any catalyst or additive, compared with the DBU catalyzed reaction in dichloromethane solution at low temperature. Mechanistic aspects of the reaction under both the conditions are discussed. In this respect, vibrational (VCD) and electronic circular dichroism (ECD) and optical rotatory dispersion (ORD) experiments proved to be valuable tools for determining the absolute configurations of the reaction products.


Miocene Fossils Reveal Ancient Roots for New Zealand's Endemic Mystacina (Chiroptera) and Its Rainforest Habitat.
The New Zealand endemic bat family Mystacinidae comprises just two Recent species referred to a single genus, Mystacina. The family was once more diverse and widespread, with an additional six extinct taxa recorded from Australia and New Zealand. Here, a new mystacinid is described from the early Miocene (19-16 Ma) St Bathans Fauna of Central Otago, South Island, New Zealand. It is the first pre-Pleistocene record of the modern genus and it extends the evolutionary history of Mystacina back at least 16 million years. Extant Mystacina species occupy old-growth rainforest and are semi-terrestrial with an exceptionally broad omnivorous diet. The majority of the plants inhabited, pollinated, dispersed or eaten by modern Mystacina were well-established in southern New Zealand in the early Miocene, based on the fossil record from sites at or near where the bat fossils are found. Similarly, many of the arthropod prey of living Mystacina are recorded as fossils in the same area. Although none of the Miocene plant and arthropod species is extant, most are closely related to modern taxa, demonstrating potentially long-standing ecological associations with Mystacina.


Simulations of chlorophyll fluorescence incorporated into the Community Land Model version 4.
Several studies have shown that satellite retrievals of solar-induced chlorophyll fluorescence (SIF) provide useful information on terrestrial photosynthesis or gross primary production (GPP). Here, we have incorporated equations coupling SIF to photosynthesis in a land surface model, the National Center for Atmospheric Research Community Land Model version 4 (NCAR CLM4), and have demonstrated its use as a diagnostic tool for evaluating the calculation of photosynthesis, a key process in a land surface model that strongly influences the carbon, water, and energy cycles. By comparing forward simulations of SIF, essentially as a byproduct of photosynthesis, in CLM4 with observations of actual SIF, it is possible to check whether the model is accurately representing photosynthesis and the processes coupled to it. We provide some background on how SIF is coupled to photosynthesis, describe how SIF was incorporated into CLM4, and demonstrate that our simulated relationship between SIF and GPP values are reasonable when compared with satellite (Greenhouse gases Observing SATellite; GOSAT) and in situ flux-tower measurements. CLM4 overestimates SIF in tropical forests, and we show that this error can be corrected by adjusting the maximum carboxylation rate (Vmax ) specified for tropical forests in CLM4. Our study confirms that SIF has the potential to improve photosynthesis simulation and thereby can play a critical role in improving land surface and carbon cycle models.


Canine leishmaniosis in Cyprus due to Leishmania infantum MON 1.
During a serological survey in 1996, a total of 601 dogs (group I) distributed all over the government controlled southern area of Cyprus was examined using an enzyme-linked immunosorbent assay (ELISA) for the presence of specific antibodies directed against soluble antigens of promastigote stages of Leishmania infantum. The overall seroprevalence rate in this group was 1.7%. A second group (group II) of dogs was selected from regions where seropositive dogs where determined within the first group. In group II specific anti-Leishmania antibodies could be detected in 30 of 301 dogs investigated (10%). The highest seroprevalence rates were found in the regions of Agios Georgios (26.2%) and Limnatis (12.2%). Ten parasite isolates from ten dogs (six with typical symptoms of canine leishmaniosis and four without symptoms) originating from five locations could be characterised by zymodeme analysis. All ten isolates were identified as L. infantum zymodeme MON 1.


Finite element modeling of optic nerve head biomechanics.
Biomechanical factors have been implicated in the development of glaucomatous optic neuropathy, particularly at the level of the lamina cribrosa. The goal of this study was to characterize the biomechanics of the optic nerve head using computer modeling techniques. Several models of the optic nerve head tissues (pre- and postlaminar neural tissue, lamina cribrosa, central retinal vessel, sclera, and pia mater) were constructed. Stresses, deformations, and strains were computed using finite element modeling for a range of normal and elevated intraocular pressures. Computed retinal surface deformations were compared with measured deformation patterns in enucleated human eyes. A sensitivity analysis was performed in which tissue properties and selected geometric features were varied. Acute IOP-induced deformation of the vitreoretinal interface was highly dependent on optic cup shape but showed a characteristic "W-shaped" profile that did not match the deformation of the anterior surface of the lamina cribrosa. The central retinal vasculature had surprisingly little effect on optic nerve head biomechanics. At an IOP of 50 mm Hg, strains (fractional elongation) in the lamina cribrosa averaged 4% to 5.5%, dependent on model geometry, with maximum strains up to 7.7%. Strains in the lamina cribrosa were more dependent on scleral stiffness, scleral thickness, and scleral canal diameter than on lamina cribrosa stiffness and optic cup shape. Computed levels of strain in the lamina cribrosa are biologically significant and capable of contributing to the development of glaucomatous optic neuropathy, even without considering the probable accentuating effect of the lamina cribrosa's microarchitecture. Depending on optic cup shape, IOP-induced deformation of the vitreoretinal interface may not match lamina cribrosa deformation. This finding implies that scanning laser tomography has limited ability to estimate lamina cribrosa deformation when imaging the anterior topography of the optic nerve head. Biomechanical effects in the lamina cribrosa depend strongly on scleral properties.


A RA-dependent, tumour-growth suppressive transcription complex is the target of the PML-RARalpha and T18 oncoproteins.
PML and Tif1a are fused to RARA and Braf, respectively, resulting in the production of PML-RARalpha and Tif1alpha-B-Raf (T18) oncoproteins. Here we show that PML, Tif1alpha and RXRalpha/RARalpha function together in a transcription complex that is dependent on retinoic acid (RA). We found that PML acts as a ligand-dependent coactivator of RXRalpha/RARalpha. PML interacts with Tif1alpha and CBP. In Pml-/- cells, the RA-dependent induction of genes such as RARB2 and the ability of Tif1alpha and CBP to act as transcriptional coactivators on RA are impaired. We show that both PML and Tif1alpha are growth suppressors required for the growth-inhibitory activity of RA. T18, similar to PML-RARalpha, disrupts the RA-dependent activity of this complex in a dominant-negative manner resulting in a growth advantage. Our data define a new pathway for the control of cell growth and tumorigenesis, and provide a new model for the pathogenesis of acute promyelocytic leukaemia (APL).


Carrier lifetime in exfoliated few-layer graphene determined from intersubband optical transitions.
We report a femtosecond transient spectroscopy study in the near to middle infrared range, 0.8-0.35 eV photon energy, on graphene and few layer graphene single flakes. The spectra show an evolving structure of photoinduced absorption bands superimposed on the bleaching caused by Pauli blocking of the interband optically coupled states. Supported by tight-binding model calculations, we assign the photoinduced absorption features to intersubband transitions as the number of layers is increased. Interestingly, the intersubband photoinduced resonances show a longer dynamics than the interband bleaching, because of their independence from the absolute energy of the carriers with respect to the Dirac point. The dynamic of these intersubband transitions reflects the lifetime of the hot carriers and provides an elegant method to access it in this important class of semimetals.


U.S. Dental Schools' Preparation for the Integrated National Board Dental Examination.
An Integrated National Board Dental Examination (INBDE) combining basic, behavioral, and clinical sciences will be implemented in 2020 to replace the current two-part National Board Dental Examination required for all candidates who seek to practice dentistry in the U.S. The aims of this study were to determine how U.S. dental schools are preparing for implementation of the INBDE and to assess their top administrators' attitudes about the new exam. A total of 150 deans, academic deans, and other administrators at all 64 U.S. dental schools with graduating classes in 2016 were emailed a 19-question electronic survey. The survey questions addressed the respondents' level of support, perceived benefits and challenges, and planned preparation strategies for the INBDE. The individual response rate was 59%, representing 57 of the 64 schools. Approximately 60% of the respondents either agreed or strongly agreed that they support the integrated exam, while roughly 25% either somewhat or strongly disagreed. While most respondents (72%) reported that their institutions would be prepared for the INBDE, 74% reported that the merged exam created additional strain for their institutions. Respondents reported viewing content integration and clinical applicability as benefits of the INBDE, while required curriculum changes and student preparedness and stress were seen as challenges. Most of the respondents reported their schools were currently employing strategies to prepare for the INBDE including meetings with faculty and students and changes to curricula and course content. The beginning of the fourth year and the end of the third year were the most frequently reported times when schools planned to require students to take the INBDE, although almost half of the respondents did not yet know what it would be required at their school. Several schools were reconsidering using the boards as a passing requirement. This study found that support for the INBDE was not universal, but strategies are under way to prepare students, faculty, and curricula for this new means of assessment.


Systemic interleukin-6 concentrations in patients with perimesencephalic non-aneurysmal subarachnoid hemorrhage.
Patients with spontaneous non-aneurysmal subarachnoid hemorrhage (non-aSAH) are considered to have a benign illness in contrast to patients with aSAH. The occurrence of the systemic inflammatory response syndrome has been linked to worse outcomes in patients with aSAH. We analyzed systemic interleukin (IL)-6, a proinflammatory cytokine, to determine whether its concentration differs between patients with non-aSAH and those with aSAH, reflecting the more benign illness. Daily systemic IL-6 levels were measured in the acute phase in 11 patients with non-aneurysmal perimesencephalic SAH (pmSAH), with bleeding strictly located around the midbrain, and in nine patients with non-aneurysmal non-perimesencephalic (non-pmSAH), with hemorrhage extending into adjacent cisterns (group 1). IL-6 levels were compared with those from patients suffering from aSAH with cerebral vasospasm (CVS) (group 2) and without CVS (group 3). The mean IL-6 level (±standard error of the mean) was significantly lower in group 1 compared to group 2 (9.9±1.9 vs. 29.1±6.7 pg/mL, p=0.018). The difference in mean IL-6 level between group 1 and 3 fell short of significance (9.9±1.9 vs. 14.9±1.1 pg/mL, p=0.073). Patients in group 1 had a significantly better outcome (Glasgow Outcome Scale score 4-5) compared to group 2 (p<0.001) and a trend towards better outcome compared to group 3 (p=0.102). A subgroup analysis revealed a higher mean IL-6 concentration in patients with non-pmSAH compared to patients with pm-SAH (p=0.001). We concluded that systemic IL-6 concentration reflects the severity of the inflammatory stress response and course of the illness. The more benign illness and good prognosis of patients with pmSAH or non-pmSAH in contrast to patients with aSAH is reflected by the lower concentrations of IL-6.


Charting the practical dimensions of understaffing from a managerial perspective: The everyday shape of the UK's sonographer shortage.
Across the last two decades, ultrasound services in many healthcare sectors have become increasingly pressurised as a consequence of upsurging demand and difficulties in recruiting viable clinicians. Indeed by 2013, the UK government's Migration Advisory Committee had listed sonography as an official 'shortage specialty'. Comparatively little research has to date, however, explored the impacts of this situation upon the departments themselves, and the individuals working therein. The core purpose of this study is, thus, to lend qualitative depth to current understandings of the frontline situation in the UK's ultrasound units, many of which are understaffed, from the perspective of their managers. Using a thematic analysis informed by a Straussian model of Grounded Theory, N = 20 extended accounts provided by ultrasound department leads in public (n = 18) and private (n = 2) units were explored. Four global themes emerged from the analysis of which the first two (the broadly sociological matters) are described in this paper. Theme 1 addresses how a lack of staff in the broader ultrasound economy has created a troublesome migratory system in contemporary UK ultrasound. Theme 2 addresses how this economy works chiefly to the advantage of the most junior and the most senior clinicians, often leaving mid-career professionals in the borderline impossible situation of having to concurrently occupy both junior and senior roles. The findings ideally open up debate on some key practical contingencies of the UK's sonographer shortage, and reflect upon literature regarding the nuanced aspects of a shifting healthcare workplace constitution.


Oil palm and gendered time use: A mixed-methods case study from West Kalimantan, Indonesia.
Measuring the social impact of oil palm requires the use of multiple metrics which capture different dimensions of well-being. To date, most studies have examined welfare outcomes at the household level, relying on a relatively narrow range of indicators. There is a need for a more diverse range of metrics to measure the social impacts of oil palm as well as more explicit accounting for study context and gendered effects. Here we demonstrate the utility of specialised time use methods used in combination with qualitative research to understand intra-household labour dynamics associated with oil palm adoption. We use a mixed-methods approach to investigate the role of smallholder oil palm plasma schemes on men and women's time use in Kapuas Hulu District, West Kalimantan. Time allocation is an important determinant of well-being as well as maternal and child nutrition and an indicator of women's empowerment and gender equality. We integrate the results from a fractional multinomial logistic regression of data from 603 individuals with qualitative findings on the subjective experience of time allocation, as well as, the causes, consequences and coping strategies to manage trade-offs in time allocation. We find that relative to non-oil-palm adopting swidden farmers, participation in oil palm plasma schemes is associated with more time spent in productive labour for both men and women, driven by off-farm labour on oil palm plantations. For women, increased time comes at the cost of reduced time spent in rest, leisure and sleep. Increased time spent in off-farm labour drives households to adapt agricultural production methods, changing cash crop production as well as accelerating swidden transitions. These changes alter gender dynamics and responsibilities within the household. Our results suggest that changes in time allocation may have significant consequences for women's well-being and gender equity. Women in the oil palm site experienced greater stress over time scarcity and employed coping strategies more frequently. Our findings indicate that time allocation could be used as an indicator of the effects of oil palm expansion and adoption on well-being and that potential effects of time scarcity on well-being, gender equity, and maternal and child nutrition should be considered by policy makers when making land use decisions.


Prophylactic oophorectomy versus screening: psychosocial outcomes in women at increased risk of ovarian cancer.
This study investigated the psychosocial outcome of prophylactic oophorectomy versus regular screening in women at increased risk of ovarian cancer. Women who had undergone prophylactic oophorectomy (n=29) were compared with women who remained on the ovarian screening programme (n=28). Assessments were made retrospectively by postal questionnaire. The surgical group showed significantly poorer functioning on two sub-scales of the Short Form (SF)-36 Health Status Questionnaire (role-emotional (p=0.04) and social functioning (p=0.01)), and there was a trend (p=0.06) for them to report more menopausal symptoms. General Health Questionnaire (GHQ) scores were significantly higher (p=0.03) in the surgical group. There were no significant differences between the groups for cancer worry or sexual functioning. Experience of the operation was better (p=0.01) and incidence of self-reported post-operative problems was lower (p=0.02) for women who had undergone the 'keyhole' rather than an 'open' procedure. Being pre-menopausal at the time of surgery predicted higher GHQ (p=0.04) and longer subjective recovery time (p=0.04). Women who have undergone prophylactic oophorectomy may have more physical and emotional symptoms than women who remain on an ovarian cancer screening programme, and may report equivalent levels of cancer worry. Those who are pre-menopausal at the time of the operation may be particularly vulnerable to psychological distress and take longer to recover post-operatively. A larger prospective study is needed to evaluate the casual versus causal role of oophorectomy in these findings, and the extent to which it allays patients' fear of cancer.


Experimental Determination of Partitioning in the Fe-Ni System for Applications to Modeling Meteoritic Metals.
Experimental trace element partitioning values are often used to model the chemical evolution of metallic phases in meteorites, but limited experimental data were previously available to constrain the partitioning behavior in the basic Fe-Ni system. In this study, we conducted experiments that produced equilibrium solid metal and liquid metal phases in the Fe-Ni system and measured the partition coefficients of 25 elements. The results are in good agreement with values modeled from IVB iron meteorites and with the limited previous experimental data. Additional experiments with low levels of S and P were also conducted, to help constrain the partitioning behaviors of elements as a function of these light elements. The new experimental results were used to derive a set of parameterization values for element solid metal-liquid metal partitioning behavior in the Fe-Ni-S, Fe-Ni-P, and Fe-Ni-C ternary systems at 0.1 MPa. The new parameterizations require that the partitioning behaviors in the light-element-free Fe-Ni system are those determined experimentally by this study, in contrast to previous parameterizations that allowed this value to be determined as a best-fit parameter. These new parameterizations, with self-consistent values for partitioning in the end-member Fe-Ni system, provide a valuable resource for future studies that model the chemical evolution of metallic phases in meteorites.


Preoperative assessment of the aging patient.
The issues related to preoperative evaluation and facial analysis for patients undergoing facial rejuvenation procedures are discussed in this article. A key component of the preoperative evaluation begins with a thorough understanding of the patient's concerns and desires for improvements. Other components of the evaluation should include a detailed assessment of the patient's medical and psychiatric history, informed consent, and photographic documentation. Additionally, we discuss facial changes associated with aging, paying special attention to some of the differences that occur between male and female patients. Different aesthetic scales and their applications in facial plastic surgery are also presented. Finally, we consider the process of facial analysis in the context of common facial plastic procedures including rhytidectomy, blepharoplasty, browlift, and minimally invasive techniques.


Sensitivity and sensitisation in relation to pain severity in knee osteoarthritis: trait or state?
It is not clear whether heightened pain sensitivity in knee osteoarthritis (OA) is related to sensitisation induced by nociceptive input from OA pathology ('state') versus other confounding factors. Conversely, some individuals may be predisposed to sensitisation irrespective of OA ('trait'). The Multicenter Osteoarthritis Study is a longitudinal cohort of persons with or at risk of knee OA. We obtained knee X-rays, pain questionnaires and comprehensive assessment of factors that can influence pain sensitivity. We examined the relation of sensitisation and sensitivity assessed by mechanical temporal summation (TS) and pressure pain thresholds (PPTs) to knee OA and knee pain severity. To test whether sensitisation and sensitivity is a 'state' induced by OA pathology, we examined the relation of PPT and TS to knee OA duration and severity. In 2126 subjects (mean age 68, mean body mass index (BMI) 31, 61% female), PPT and TS were not associated with radiographic OA (ORs 0.9-1.0 for PPT and TS; p>0.05). However, PPT and TS were associated with pain severity (ORs: 1.7-2.0 for PPT; 1.3-1.6 for TS; p<0.05). Knee OA duration and radiographic severity were not associated with PPT or TS. PPT and TS were associated with OA-related pain, but not radiographic OA after accounting for pertinent confounders in this large cohort. Lack of association with disease duration suggests at least some sensitisation and pain sensitivity may be a trait rather than state. Understanding the relationship between pathological pain and pain sensitivity/sensitisation offers insight into OA pain risk factors and pain management opportunities.


Telomere and LINE-like elements at the termini of the Chlorella chromosome I.
The telomeres of Chlorella chromosomes consisted of 5'-TTTAGGG repeats, which are exactly the same as those of higher plants. This sequence was reiterated approximately 70 times at both termini of chromosome I. Subtelomeric sequences next to the telomeres were totally different between the right and left arms. On the left-side subtelomeric region, polyA associated LINE (long interspersed element)-like elements were found tandemly repeated just next to the telomeric repeats. It is very interesting to compare this unique structure with telomeres of Drosophila, where a transposable element play a major role in forming telomerase-generated repeats. We propose a mechanism of transposon-mediated healing of a broken chromosomal end that would operate in the unicellular green alga Chlorella.


The impact of atypical employment on individual wellbeing: evidence from a panel of British workers.
This study explores the relationship between individual wellbeing and atypical employment, which includes both temporary and part-time employment schemes. Individual wellbeing is measured in terms of subjective indicators of mental health, general health status, life satisfaction, and job satisfaction. It addresses four questions: (1) Are workers on a temporary contract more likely to report poor health and poor life and job satisfaction than those who are employed in permanent jobs? (2) Is this the case for part-time workers compared to those who are in a full-time job? (3) Do changes in employment profiles (e.g., from a fixed-term contract to a permanent job, or from part-time employment to full-time employment) affect individuals' health and life satisfaction? (4) Are there differences in such relationships between men and women? To answer these questions, logistic regression models were used to analyse a panel of almost 7000 male and female workers from the first 10 waves of the British Household Panel Survey, 1991-2000. Controlling for background characteristics, atypical employment does not appear to be associated with adverse health consequences for either men or women, when both health and employment are measured at the same time. However, there is evidence that job satisfaction is reduced for seasonal/casual workers and is higher for part-timers. Taking account of selection issues does not change the general picture: the chances of poor mental and physical health and low life satisfaction are unaffected by atypical employment and some of the effects of job satisfaction persist. In addition, very few employment transitions appear to be consequential for a worsening in health outcomes, which tends to be observed in the case of job satisfaction. Although the pattern of results suggests that atypical forms of employment do not have durable adverse health consequences on workers, public policies that aim at improving the working conditions of workers in weak bargaining positions should give special attention to equity issues, including the possible health effects of experience of work in atypical employment arrangements.


Vigorous lateral export of the meltwater outflow from beneath an Antarctic ice shelf.
The instability and accelerated melting of the Antarctic Ice Sheet are among the foremost elements of contemporary global climate change. The increased freshwater output from Antarctica is important in determining sea level rise, the fate of Antarctic sea ice and its effect on the Earth's albedo, ongoing changes in global deep-ocean ventilation, and the evolution of Southern Ocean ecosystems and carbon cycling. A key uncertainty in assessing and predicting the impacts of Antarctic Ice Sheet melting concerns the vertical distribution of the exported meltwater. This is usually represented by climate-scale models as a near-surface freshwater input to the ocean, yet measurements around Antarctica reveal the meltwater to be concentrated at deeper levels. Here we use observations of the turbulent properties of the meltwater outflows from beneath a rapidly melting Antarctic ice shelf to identify the mechanism responsible for the depth of the meltwater. We show that the initial ascent of the meltwater outflow from the ice shelf cavity triggers a centrifugal overturning instability that grows by extracting kinetic energy from the lateral shear of the background oceanic flow. The instability promotes vigorous lateral export, rapid dilution by turbulent mixing, and finally settling of meltwater at depth. We use an idealized ocean circulation model to show that this mechanism is relevant to a broad spectrum of Antarctic ice shelves. Our findings demonstrate that the mechanism producing meltwater at depth is a dynamically robust feature of Antarctic melting that should be incorporated into climate-scale models.


Ketamine administered to pregnant rats in the second trimester causes long-lasting behavioral disorders in offspring.
Commonly used anesthetic agents, e.g. ketamine, may be neurotoxic to the developing brain but there has been little attention to the neurobehavioral consequences for offspring when used for maternal anesthesia. We hypothesize that treatment of pregnant rats with ketamine during the second trimester would affect brain development of the offspring. Pregnant rats on gestational day 14, about equal to midtrimester pregnancy in humans, received a sedative dose of ketamine intravenously for 2h. Brain hippocampal morphology of their pups at postnatal days 0 (P0) and P30 was examined by Nissl-staining and the characteristics of dendrites were determined using the Golgi-Cox staining, while cell proliferation in subventricular zone (SVZ) and dentate gyrus (DG) was labeled with bromodeoxyuridine (BrdU). Their neurobehavioral functions were tested at P25-30 after which the NR1 and NR2 subunits of N-methyl-d-aspartate (NMDA) receptor, brain-derived neurotrophic factor (BDNF) and postsynaptic density protein 95 (PSD-95) in the hippocampus were analyzed by western blot. When pregnant rats were exposed to ketamine, there was neuronal loss, pyramidal neuronal abnormality and reduced cell proliferation in the hippocampus of offspring. These morphological abnormalities were associated with depression- and anxiety-like behaviors, and impaired memory up to young adult age. The treatment further caused NR2A receptor subunit up-regulation and NR2B receptor subunit, BDNF and PSD-95 down-regulation. These data suggest that maternal anesthesia with ketamine during the fetal brain development period can cause fetal brain damage and subsequent neurobehavioral abnormality, which is likely associated with the imbalanced expression of NMDA receptor subunits.