Discriminatory processing of the precursor forms of cytochrome P-450scc and adrenodoxin by adrenocortical and heart mitochondria.
The mitochondrial proteins involved in adrenocortical steroidogenesis are synthesized as higher molecular weight precursors which require processing by the mitochondria to their mature sizes. The post-translational maturation of two of these proteins has been examined: the cholesterol side chain cleavage cytochrome P-450 (P-450scc) and the iron-sulfur protein, adrenodoxin. Total translation products synthesized in a cell-free system programmed by bovine adrenocortical poly(A+) RNA were incubated with isolated bovine adrenocortical or heart mitochondria followed by immunoisolation of radiolabeled P-450scc or adrenodoxin. In the presence of adrenocortical mitochondria, the precursor form of P-450scc was converted into a trypsin-resistant form that had the same molecular weight as mature P-450scc. Unlike adrenocortical mitochondria, heart mitochondria were unable to process the P-450scc precursor which remained unaltered and trypsin-sensitive. In addition, a matrix fraction of heart mitochondria did not cleave the P-450scc precursor. In contrast, the adrenodoxin precursor did not exhibit similar specificity as it was processed to the mature form by both adrenocortical and heart mitochondria. Also, the adrenocortical mitochondria were not restricted to processing endogenous proteins as they imported and cleaved the precursor to ornithine transcarbamylase. The results indicate that some mitochondrial precursor proteins have tertiary structures which allow them to be recognized by all mitochondria while other mitochondrial precursor proteins have structures recognizable by only specialized mitochondria.


ART adherence changes among patients in community substance use treatment: a preliminary analysis from MACH14.
Opiate substitution treatment has been associated with better adherence to lifesaving antiretroviral medications, but the impact of other substance abuse treatment on adherence is unknown. In this study, 215 patients who had been in adherence-focused research studies provided electronically-measured adherence data and a measure of whether the patient had recently been in substance abuse treatment. Recent engagement in substance abuse treatment was independently associated with significantly higher adherence, after covarying for recent substance use and other factors potentially affecting adherence. The findings suggest that substance abuse treatment is associated with better adherence. Potential mechanisms by which substance abuse treatment improves adherence, such as more stability or more future-orientation, require further study.


Distinct fitness costs associated with the knockdown of RNAi pathway genes in western corn rootworm adults.
RNA interference (RNAi) based approaches can potentially be used to control insect pests. These approaches may depend on the usage of microRNA (miRNA) or double stranded RNA (dsRNA) mediated gene knockdown, which likely involves proteins that regulate these pathways, such as Argonaute 1 (Ago1), Argonaute 2 (Ago2), Dicer 1 (Dcr1), Dicer 2 (Dcr2), and Drosha in insects. We previously performed functional characterization of Ago2 and Dcr2 of western corn rootworm (WCR), Diabrotica virgifera virgifera (Coleoptera: Chrysomelidae) and observed that knockdown of Ago2 and Dcr2 ameliorated the lethal effect induced by the dsRNA-mediated knockdown of an essential gene in WCR, thereby confirming the involvement of Ago2 and Dcr2 in the dsRNA pathway. In the current study, we identified and characterized additional members of the Argonaute and Dicer gene families, namely Ago1, Ago3, Aubergine, and Dcr1, in a previously developed WCR transcriptome. We also identified a Drosha homolog in the same transcriptome. We evaluated the impacts on WCR adult fitness associated with the dsRNA-mediated knockdown of Ago1, Ago2, Dcr1, Dcr2, and Drosha genes. Among these putative RNAi pathway genes, only the knockdown of Ago1 incurred significant fitness costs such as reduced survival and oviposition rate, as well as decreased egg viability. The present study, to our knowledge, represents the first report showing that Ago1 is critical to the survival of insect adults. Our findings suggest that Ago1 plays an essential role in broader life stages of an insect than previously thought. Importantly, since fitness costs were not observed, downregulation or loss of function of RNAi pathway genes such as Ago2 or Dcr2 may confer resistance to pest control measures that rely on the normal functions of these genes. However, the precise roles of these genes under field conditions (i.e., in the presence of possible viral pathogens) requires further investigation.


Histidine 55 of tryptophan 2,3-dioxygenase is not an active site base but regulates catalysis by controlling substrate binding.
Tryptophan 2,3-dioxygenase (TDO) from Xanthomonas campestris is a highly specific heme-containing enzyme from a small family of homologous enzymes, which includes indoleamine 2,3-dioxygenase (IDO). The structure of wild type (WT TDO) in the catalytically active, ferrous (Fe (2+)) form and in complex with its substrate l-tryptophan ( l-Trp) was recently reported [Forouhar et al. (2007) Proc. Natl. Acad. Sci. U.S.A. 104, 473-478] and revealed that histidine 55 hydrogen bonds to l-Trp, precisely positioning it in the active site and implicating it as a possible active site base. In this study the substitution of the active site residue histidine 55 by alanine and serine (H55A and H55S) provides insight into the molecular mechanism used by the enzyme to control substrate binding. We report the crystal structure of the H55A and H55S mutant forms at 2.15 and 1.90 A resolution, respectively, in binary complexes with l-Trp. These structural data, in conjunction with potentiometric and kinetic studies on both mutants, reveal that histidine 55 is not essential for turnover but greatly disfavors the mechanistically unproductive binding of l-Trp to the oxidized enzyme allowing control of catalysis. This is demonstrated by the difference in the K d values for l-Trp binding to the two oxidation states of wild-type TDO (3.8 mM oxidized, 4.1 microM reduced), H55A TDO (11.8 microM oxidized, 3.7 microM reduced), and H55S TDO (18.4 microM oxidized, 5.3 microM reduced).


Early results of the Bernese periacetabular osteotomy for symptomatic dysplasia in Charcot-Marie-Tooth disease.
Charcot-Marie-Tooth disease (CMTD) is one of the most common inherited neurologic disorders and can be associated with hip dysplasia. Little is known regarding outcomes of the PAO for patients with CMTD. Our purpose is to document the early results and complications of the PAO for hip dysplasia associated with CMTD. A two centre, retrospective clinical and radiographic review was performed. Demographic and surgical data were recorded. Pre- and postoperative lateral centre edge angle (LCEA), acetabular index (AI), ventral centre edge angle (VCEA), and Tönnis osteoarthritis grade were compared. Hips were classified according to Severin. The Harris Hip Score (HHS) and the Western Ontario and McMasters University (WOMAC) index documented self-reported function. Nineteen hips in 14 patients underwent PAO, mean age 16.2 (range 11.2-21 years). Thirteen concomitant procedures were performed, including seven proximal femoral osteotomies. Average follow-up was 3.4 years (range 0.9-8.5). Postoperative radiographic measurements significantly improved. Complications included femoral head AVN, transient complete bilateral peroneal nerve palsy, inferior rami fractures, and heterotopic ossification (Brooker stage 3). The HHS significantly improved from a mean 49.6 preoperatively to 82.2 at final follow-up of four patients. Seven subjects reported a mean postoperative WOMAC score of 94 (range 58.3-100). Most patients presented with severe dysplasia in the second decade of life. The PAO successfully corrected the radiographic abnormalities. Complications were common. The majority of patients reported improved outcomes, although seven showed signs of radiographic progression of osteoarthritis.


Clinical relevance of cytomegalovirus viraemia(*,†).
Using new sensitive quantitative polymerase chain reaction (PCR) assays, cytomegalovirus (CMV) DNA is often detectable in the plasma of immunosuppressed patients. We investigated the prognostic value of a positive CMV DNA test for the development of CMV end-organ disease, other AIDS-defining events and mortality. A survival analysis was performed, using the Kaplan-Meier method and Cox proportional hazards models, for patients prospectively followed in the Swiss HIV Cohort Study, from January 1996 to December 2007, who were CMV-seropositive, had a CD4 count of ≤ 100 cells/μL, and had a plasma sample available for the measurement of baseline CMV DNA with an ultrasensitive PCR. The outcome analysed was an AIDS-defining event, including CMV end-organ disease, or death. Variables analysed at the time of CMV measurement were demographic variables, CD4 cell counts, HIV-1 RNA loads, and use and type of highly active antiretroviral therapy (HAART). Of 1128 patients, 208 (18%) presented an AIDS-defining event and 246 (22%) died. A total of 368 patients (34% of samples) had detectable CMV DNA at baseline, with DNA concentrations of up to 38 800 copies/mL. In the multivariate analysis, CMV DNA predicted evolution not only towards CMV end-organ disease [hazard ratio (HR) 12.6; 95% confidence interval (CI) 4.27-37.41], but also towards other AIDS-defining events (HR 2.6; 95% CI 1.60-4.33) and death (HR 1.9; 95% CI 1.10-3.34). Quantitative CMV DNA detected in the plasma of HIV-infected patients with CD4 counts ≤ 100 cells/μL is a predictor for HIV disease progression, CMV disease and death. A single low value of 80 copies/mL identifies patients at low but significantly increased risk during the following months, after the measurement.


General sexual desire, but not desire for uncommitted sexual relationships, tracks changes in women's hormonal status.
Several recent longitudinal studies have investigated the hormonal correlates of both young adult women's general sexual desire and, more specifically, their desire for uncommitted sexual relationships. Findings across these studies have been mixed, potentially because each study tested only small samples of women (Ns = 43, 33, and 14). Here we report results from a much larger (N = 375) longitudinal study of hormonal correlates of young adult women's general sexual desire and their desire for uncommitted sexual relationships. Our analyses suggest that within-woman changes in general sexual desire are negatively related to progesterone, but are not related to testosterone or cortisol. We observed some positive relationships for estradiol, but these were generally only significant for solitary sexual desire. By contrast with our results for general sexual desire, analyses showed no evidence that changes in women's desire for uncommitted sexual relationships are related to their hormonal status. Together, these results suggest that changes in hormonal status contribute to changes in women's general sexual desire, but do not influence women's desire for uncommitted sexual relationships.


Anti-inflammatory strategies in hypertension: focus on COX-1 and COX-2.
An increasing body of evidence suggests that elevated levels of blood pressure may induce a proinflammatory response. Indeed, both C-reactive protein and blood pressure are independent determinants of cardiovascular risk, and, in combination, each parameter has additional predictive value. Hence, strategies targeted to lower blood pressure and reduce vascular inflammation may potentially provide clinical benefit. In this review, we discuss the role of chronic low-grade inflammation in the context of cardiovascular disease with a focus on roles of cyclooxygenase-1 and -2 in potential anti-inflammatory treatment strategies.


Mu opioid receptor-effector coupling and trafficking in dorsal root ganglia neurons.
Morphine induces profound analgesic tolerance in vivo despite inducing little internalization of the mu opioid receptor (muOR). Previously proposed explanations suggest that this lack of internalization could either lead to prolonged signaling and associated compensatory changes in downstream signaling systems, or that the receptor is unable to recycle and resensitize and so loses efficacy, either mechanism resulting in tolerance. We therefore examined, in cultured neurons, the relationship between muOR internalization and desensitization in response to two agonists, D-Ala2, N-MePhe4, Gly5-ol-enkephalin (DAMGO) and morphine. In addition, we studied the chimeric mu/delta opioid receptor (mu/ partial differentialOR) which could affect internalization and desensitization in neurons. Dorsal root ganglia neurons from muOR knockout mice were transduced with an adenovirus expressing either receptor and their respective internalization, desensitization and trafficking profiles determined. Both receptors desensitized equally, measured by Ca2+ current inhibition, during the first 5 min of agonist exposure to DAMGO or morphine treatment, although the mu/partial differentialOR desensitized more extensively. Such rapid desensitization was unrelated to internalization as DAMGO, but not morphine, internalized both receptors after 20 min. In response to DAMGO the mu/partial differentialOR internalized more rapidly than the muOR and was trafficked through Rab4-positive endosomes and lysosomal-associated membrane protein-1-labeled lysosomes whereas the muOR was trafficked through Rab4 and Rab11-positive endosomes. Chronic desensitization of the Ca2+ current response, after 24 h of morphine or DAMGO incubation, was seen in the DAMGO, but not morphine-treated, muOR-expressing cells. Such persistence of signaling after chronic morphine treatment suggests that compensation of downstream signaling systems, rather than loss of efficacy due to poor receptor recycling, is a more likely mechanism of morphine tolerance in vivo. In contrast to the muOR, the mu/partial differentialOR showed equivalent desensitization whether morphine or DAMGO treated, but internalized further with DAMGO than morphine. Such ligand-independent desensitization could be a result of the observed higher rate of synthesis and degradation of this chimeric receptor.


Hereditary diffuse gastric cancer: implications of genetic testing for screening and prophylactic surgery.
Approximately 10% of patients with gastric cancer show familial clustering, and 3% show autosomal dominance and high penetrance. Hereditary diffuse gastric cancer (HDGC) is an autosomal-dominant, inherited cancer syndrome in which affected individuals develop diffuse-type gastric cancer at a young age. Inactivating mutations in the E-cadherin gene CDH1 have been identified in 30% to 50% of patients. CDH1 mutation carriers have an approximately 70% lifetime risk of developing DGC, and affected women carry an additional 20% to 40% risk of developing lobular breast cancer. Because endoscopic surveillance is ineffective in identifying early HDGC, gene-directed prophylactic total gastrectomy currently is offered for CDH1 mutation carriers. In series of asymptomatic individuals undergoing total gastrectomy for CDH1 mutations, the removed stomachs usually contain small foci of early DGC, making surgery not prophylactic but curative. The authors of this review recommend consideration of total gastrectomy in CDH1 mutation carriers at an age 5 years younger than the youngest family member who developed gastric cancer. Individuals who choose not to undergo prophylactic gastrectomy should be followed with biannual chromoendoscopy, and women with CDH1 mutations also should undergo regular surveillance with magnetic resonance imaging studies of the breast. Because of the emergence of gene-directed gastrectomy for HDGC, today, a previously lethal disease is detected by molecular techniques, allowing curative surgery at an early stage.


Acute, sublethal, and combination effects of azadirachtin and Bacillus thuringiensis on the cotton bollworm, Helicoverpa armigera.
The cotton bollworm, Helicoverpa armigera Hübner (Lepidoptera: Noctuidae) is a polyphagous and cosmopolitan insect pest that causes damage to various plants. In this study, the lethal and sublethal effects of azadirachtin and Bacillus thuringiensis Berliner sub sp . kurstaki (Bacillales: Bacillaceae) were evaluated on third instar H. armigera under laboratory conditions. The LC50 values of azadirachtin and Bt were 12.95 and 96.8 µg a.i./mL, respectively. A total mortality of 56.7% was caused on third instar larvae when LC20 values of the insecticides were applied in combination with each other. The LT50 values of azadirachtin and Bt were 4.8 and 3.6 days, respectively. The results of the sublethal study showed that the application of LC30 value of azadirachtin and Bt reduced the larval and pupal weight and increased larval and pupal duration of H. armigera. The longevity and fecundity of female adults were affected significantly by the insecticides. Female fecundity was reduced by the treatments, respectively. The lowest adult emergence ratio and pupation ratio were observed in the azadirachtin treatment. The results indicated that both insecticides have high potential for controlling of the pest.


Femoral and sciatic nerve blockades and incision site infiltration in rabbits undergoing stifle joint arthrotomy.
This study was designed to determine whether perineural injections of local anaesthetics decreases intraoperative nociception and improves postoperative analgesia in New Zealand White rabbits undergoing experimental stifle arthrotomy. All animals were anaesthetized using isoflurane and received morphine intramuscularly. The sciatic and femoral nerves of the leg to be operated on were located using a nerve stimulator (1 Hz, 0.5 mA). Rabbits were assigned to a treatment group (LB; n = 12) or a placebo group (P; n = 12) in a randomized blinded fashion. Group LB received lidocaine 2% (1 mg/kg) combined with bupivacaine 0.5% (0.25 mg/kg) injections around the sciatic and femoral nerves (0.1 mL/kg total volume per site) and subcutaneous infiltration of the incision site with lidocaine 1% (1.25 mg/kg). Group P received the same volume of 0.9% NaCl. Rabbits in group P required higher doses of intraoperative fentanyl and propofol to reduce heart rate and suppress increase in systolic blood pressure, and maintain an adequate anaesthetic plane. Interventional analgesia (buprenorphine and carprofen) was required significantly earlier in rabbits in group P (2 and 6 h after the first nerve blockade, respectively) based on assessment of their pain-related behaviour and range of motion. Using a visual analogue scale (0 mm= no pain to 100 mm= maximal possible pain), rabbits in group LB were judged to show significantly less pain compared with rabbits in group P (14 ± 10 mm and 37 ± 25 mm, respectively) 2 h after nerve blockade. In conclusion, this technique of perineural analgesia combined with incision site infiltration reduced intraoperative fentanyl requirements and improved postoperative analgesia in rabbits.


How do respiratory patients perceive oxygen therapy? A critical interpretative synthesis of the literature.
Oxygen therapy is a common intervention in health care worldwide; yet, despite universal use, it is evident through poor practice that oxygen is often prescribed and administered injudiciously. It is proposed that possibly an influencing culture presides, whereby oxygen is often poorly understood and uncertainty regarding its use exists. It is unclear where the origins of this culture lie but exploring perceptions may enlighten the problem. A review of the literature was undertaken to establish what is already known about this elusive phenomenon. The paucity of any direct evidence regarding perceptions of oxygen directed the review to utilize a critical interpretative synthesis (CIS). The aim of this study was to explore how respiratory patients perceive oxygen therapy. A systematic search in Medline, Cinahl, Embase, British Nursing Index and PsychInfo yielded 1514 studies of which 42 were selected to consider the review question. The CIS allowed evidence from across studies to synthesize existing and new interpretations of data related to patients' perceptions of oxygen therapy. Synthetic constructs then informed the synthesizing arguments, namely positive - feeling safe, enabler and comforter; negative - fear, oxygen versus self, restriction and embarrassment; and impartiality - mixed blessings. The findings are divergent, and at times contradictory. There appears uncertainty among patients regarding the purpose and benefits of oxygen therapy, though an underlying faith in health-care professionals is apparent. This faith seems to foster acceptance of a life-changing therapy, despite the impact, burden and incomplete understanding. There is a clear need for further research regarding these elusive perceptions in order to improve clinical practice in respect of oxygen.


Characterization of Charge States in Conducting Organic Nanoparticles by X-ray Photoemission Spectroscopy.
The metallic and semiconducting character of a large family of organic materials based on the electron donor molecule tetrathiafulvalene (TTF) is rooted in the partial oxidation (charge transfer or mixed valency) of TTF derivatives leading to partially filled molecular orbital-based electronic bands. The intrinsic structure of such complexes, with segregated donor and acceptor molecular chains or planes, leads to anisotropic electronic properties (quasi one-dimensional or two-dimensional) and morphology (needle-like or platelet-like crystals). Recently, such materials have been synthesized as nanoparticles by intentionally frustrating the intrinsic anisotropic growth. X-ray photoemission spectroscopy (XPS) has emerged as a valuable technique to characterize the transfer of charge due to its ability to discriminate the different chemical environments or electronic configurations manifested by chemical shifts of core level lines in high-resolution spectra. Since the photoemission process is inherently fast (well below the femtosecond time scale), dynamic processes can be efficiently explored. We determine here the fingerprint of partial oxidation on the photoemission lines of nanoparticles of selected TTF-based conductors.


Lockdown, bottoms up? Changes in adolescent substance use across the COVID-19 pandemic.
The COVID-19 pandemic notably altered adolescent substance use during the initial stage (Spring 2020) of the pandemic. The purpose of this longitudinal study is to examine trajectories of adolescent substance use across the pandemic and subsequent periods of stay-at-home orders and re-opening efforts. We further examined differences as a function of current high school student versus graduate status. Adolescents (n = 1068, 14-18 years, Mage = 16.95 years and 76.7% female at T1) completed 4 different self-report surveys, starting during the first stay-at-home order and ending approximately 14 months later. Negative binomial hurdle models predicted: (1) the likelihood of no substance use and (2) frequency of days of substance use. As hypothesized, results demonstrated significant increases in adolescents' likelihood of alcohol use, binge drinking, and cannabis use once initial stay-at-home orders were lifted, yet few changes occurred as a result of a second stay-at-home order, with rates never lowering again to that of the first lockdown. Further, graduates (and particularly those who transitioned out of high school during the study) demonstrated a greater likelihood and frequency of substance use and were more stable in their trajectories across periods of stay-at-home orders than current high school students. Unexpectedly, however, there was a strong increase in current high school students' likelihood of e-cigarette use and a significant linear increase in participants' frequency of e-cigarette use over the study. Results suggest adolescent substance use, and in particular, e-cigarette use among current high school students, may be of increasing concern as the pandemic evolves.


Drosophila patterning is established by differential association of mRNAs with P bodies.
The primary embryonic axes in flies, frogs and fish are formed through translational regulation of localized transcripts before fertilization. In Drosophila melanogaster, the axes are established through the transport and translational regulation of gurken (grk) and bicoid (bcd) messenger RNA in the oocyte and embryo. Both transcripts are translationally silent while being localized within the oocyte along microtubules by cytoplasmic dynein. Once localized, grk is translated at the dorsoanterior of the oocyte to send a TGF-α signal to the overlying somatic cells. In contrast, bcd is translationally repressed in the oocyte until its activation in early embryos when it forms an anteroposterior morphogenetic gradient. How this differential translational regulation is achieved is not fully understood. Here, we address this question using ultrastructural analysis, super-resolution microscopy and live-cell imaging. We show that grk and bcd ribonucleoprotein (RNP) complexes associate with electron-dense bodies that lack ribosomes and contain translational repressors. These properties are characteristic of processing bodies (P bodies), which are considered to be regions of cytoplasm where decisions are made on the translation and degradation of mRNA. Endogenous grk mRNA forms dynamic RNP particles that become docked and translated at the periphery of P bodies, where we show that the translational activator Oo18 RNA-binding protein (Orb, a homologue of CEPB) and the anchoring factor Squid (Sqd) are also enriched. In contrast, an excess of grk mRNA becomes localized inside the P bodies, where endogenous bcd mRNA is localized and translationally repressed. Interestingly, bcd mRNA dissociates from P bodies in embryos following egg activation, when it is known to become translationally active. We propose a general principle of translational regulation during axis specification involving remodelling of transport RNPs and dynamic partitioning of different transcripts between the translationally active edge of P bodies and their silent core.


Escalating trends in the urban metabolism of Hong Kong: 1971-1997.
Urban metabolism measures quantitatively a city's load on the natural environment. We update the Newcombe et al. (3) pioneering study of Hong Kong's urban metabolism in 1971, highlighting trends in resource consumption and waste generation. Per capita food, water and materials consumption have surged since the early 1970s by 20%, 40%, and 149%, respectively. Tremendous pollution has accompanied this growing affluence and materialism, and total air emissions, CO2 outputs, municipal solid wastes, and sewage discharges have risen by 30%, 250%, 245%, and 153%. As a result, systemic overload of land, atmospheric and water systems has occurred. While some strategies to tackle deteriorating environmental quality have succeeded, greater and more far-reaching changes in consumer behavior and government policy are needed if Hong Kong is to achieve its stated goal of becoming "a truly sustainable city" in the 21st century.


In vivo molecular and genomic imaging: new challenges for imaging physics.
The emerging and rapidly growing field of molecular and genomic imaging is providing new opportunities to directly visualize the biology of living organisms. By combining our growing knowledge regarding the role of specific genes and proteins in human health and disease, with novel ways to target these entities in a manner that produces an externally detectable signal, it is becoming increasingly possible to visualize and quantify specific biological processes in a non-invasive manner. All the major imaging modalities are contributing to this new field, each with its unique mechanisms for generating contrast and trade-offs in spatial resolution, temporal resolution and sensitivity with respect to the biological process of interest. Much of the development in molecular imaging is currently being carried out in animal models of disease, but as the field matures and with the development of more individualized medicine and the molecular targeting of new therapeutics, clinical translation is inevitable and will likely forever change our approach to diagnostic imaging. This review provides an introduction to the field of molecular imaging for readers who are not experts in the biological sciences and discusses the opportunities to apply a broad range of imaging technologies to better understand the biology of human health and disease. It also provides a brief review of the imaging technology (particularly for x-ray, nuclear and optical imaging) that is being developed to support this new field.


Cerebrospinal fluid rhinorrhea: An institutional perspective from Pakistan.
The management of cerebrospinal fluid (CSF) rhinorrhea has evolved over the last two decades. We present here a review of our 11-year data on CSF rhinorrhea and its management at a tertiary care hospital in a developing country, with particular reference to the diagnosis, surgical management and outcome of the disease. The medical charts of all patients with a diagnosis of CSF rhinorrhea over an 11-year period were reviewed. The etiology of CSF rhinorrhea was classified into three categories: spontaneous, iatrogenic and traumatic. All the patients were divided into three categories based on the type of management as conservative, intracranial and transnasal endoscopic groups. A total of 43 patients fulfilled our inclusion criteria and were included in the final analysis. Eleven of the 43 patients were managed conservatively, while 22 underwent intracranial repairs; 10 patients had transnasal endoscopic repairs. The primary success rate for the transnasal approach was 70% compared to 86% for the intracranial repair. Blood loss, special care unit (SCU) stay and total cost were found to be significantly less in the transnasal endoscopic group. Computed tomography (CT) cisternography was found to have the highest sensitivity and specificity. Further, no postoperative complications were found in the transnasal endoscopic group, while five patients from the intracranial group developed various complications. We conclude that the transnasal endoscopic approach has comparable success rates with the intracranial approach and significantly lower morbidity.


A compartmental model to quantitate in vivo glucose transport in the human forearm.
Glucose transport is a critical step in the control of glucose disposal that, until presently, has not been quantitated in vivo in humans. We have employed the perfused forearm and euglycemic insulin-clamp techniques in combination with a dual-tracer injection to measure basal and insulin-mediated glucose transport in six normal subjects. L-[3H]glucose, which is not transported, was used to trace extracellular glucose kinetics; 3-O-[14C]-methyl-D-glucose, transportable but not metabolizable, was used to monitor glucose movement across the cell membrane. After bolus intra-arterial injection of the two tracers, plasma samples were obtained every 15-30 s for 10 min from a deep forearm vein to determine the washout curves. A linear compartmental model was developed that accounts for blood flow heterogeneity. It consists of three parallel, two-compartment chains merging into the sampling compartment to which cellular compartments are appended. A priori identifiability analysis was performed. The uniquely identifiable parameterization includes the transport rate constants of glucose into and out of the cell. The model was identified using nonlinear least-squares parameter estimation. Transport parameters are estimated with very good precision, and their reproducibility is satisfactory. The model also allows the estimation of the mean arteriovenous transit times of both the extracellular and the transported tracer. The compartmental model provides a novel approach to investigate glucose transport in vivo in humans.


Distribution, determinants, and normal reference values of thoracic and abdominal aortic diameters by computed tomography (from the Framingham Heart Study).
Current screening and detection of asymptomatic aortic aneurysms is based largely on uniform cut-point diameters. The aims of this study were to define normal aortic diameters in asymptomatic men and women in a community-based cohort and to determine the association between aortic diameters and traditional risk factors for cardiovascular disease. Measurements of the diameters of the ascending thoracic aorta (AA), descending thoracic aorta (DTA), infrarenal abdominal aorta (IRA), and lower abdominal aorta (LAA) were acquired from 3,431 Framingham Heart Study (FHS) participants. Mean diameters were stratified by gender, age, and body surface area. Univariate associations with risk factor levels were examined, and multivariate linear regression analysis was used to assess the significance of covariate-adjusted relations with aortic diameters. For men, the average diameters were 34.1 mm for the AA, 25.8 mm for the DTA, 19.3 mm for the IRA, and 18.7 mm for the LAA. For women, the average diameters were 31.9 mm for the AA, 23.1 mm for the DTA, 16.7 mm for the IRA, and 16.0 mm for the LAA. The mean aortic diameters were strongly correlated (p <0.0001) with age and body surface area in age-adjusted analyses, and these relations remained significant in multivariate regression analyses. Positive associations of diastolic blood pressure with AA and DTA diameters in both genders and pack-years of cigarette smoking with DTA diameter in women and IRA diameter in men and women were observed. In conclusion, average diameters of the thoracic and abdominal aorta by computed tomography are larger in men compared with women, vary significantly with age and body surface area, and are associated with modifiable cardiovascular disease risk factors, including diastolic blood pressure and cigarette smoking.


Endoplasmic reticulum stress is a mediator of posttransplant injury in severely steatotic liver allografts.
Hepatic steatosis continues to present a major challenge in liver transplantation. These organs have been shown to have increased susceptibility to cold ischemia/reperfusion (CIR) injury in comparison with otherwise comparable lean livers; the mechanisms governing this increased susceptibility to CIR injury are not fully understood. Endoplasmic reticulum (ER) stress is an important link between hepatic steatosis, insulin resistance, and metabolic syndrome. In this study, we investigated ER stress signaling and blockade in the mediation of CIR injury in severely steatotic rodent allografts. Steatotic allografts from genetically leptin-resistant rodents had increased ER stress responses and increased markers of hepatocellular injury after liver transplantation into strain-matched lean recipients. ER stress response components were reduced by the chemical chaperone taurine-conjugated ursodeoxycholic acid (TUDCA), and this resulted in an improvement in the allograft injury. TUDCA treatment decreased nuclear factor kappa B activation and the proinflammatory cytokines interleukin-6 and interleukin-1β. However, the predominant response was decreased expression of the ER stress cell death mediator [CCAAT/enhancer-binding protein homologous protein (CHOP)]. Furthermore, activation of inflammation-associated caspase-11 was decreased, and this linked ER stress/CHOP to proinflammatory cytokine production after steatotic liver transplantation. These data confirm ER stress in steatotic allografts and implicate this as a mediating mechanism of inflammation and hepatocyte death in the steatotic liver allograft.


Gene expression analysis in human renal allograft biopsy samples using high-density oligoarray technology.
High-density oligoarray technology is a novel method for screening the expression of thousands of genes in a small tissue sample. Oligoarray analysis of genes expressed during human renal allograft rejection has not been reported previously. Seven human renal allograft biopsies with histologic evidence of acute cellular rejection and three renal allograft biopsies without evidence of rejection (control) were analyzed for the expression of 6800 human genes using high-density oligoarrays (GeneChip, Affymetrix, Santa Clara, CA). Quantitative expression of gene transcripts was determined and a comparison analysis between acute rejection and control biopsy samples was performed. Up-regulation of a specific gene transcript during acute rejection was considered to be significant if transcript abundance increased fourfold or more relative to control biopsy samples. Comparison analysis revealed that between 32 and 219 gene transcripts are up-regulated (>fourfold) during acute rejection. Of these transcripts, only four (human monokine induced by interferon-gamma, T-cell receptor active beta-chain protein, interleukin-2 stimulated phosphoprotein, and RING4 (a transporter involved in antigen presentation)) were consistently up-regulated in each acute rejection sample relative to at least two of three control biopsy samples. Six other genes were up-regulated in six of seven acute rejection samples. These were interferon-stimulated growth factor-3, complement factor 3, nicotinamide N-methyltransferase, macrophage inflammatory protein-3beta, myeloid differentiation protein, and CD18. Only two gene transcripts were down-regulated in five of seven acute rejection samples. Significant up-regulation of cytotoxic T-cell effector molecules, previously reported as markers of acute renal rejection in humans, was not detected. High-density oligoarray technology is useful for screening gene expression in transplanted tissues undergoing acute rejection. Because this method does not rely on a priori knowledge of which genes are involved in acute rejection, it is likely to yield novel insights into the mechanisms and diagnosis of rejection.


EULAR recommendations for the management of antiphospholipid syndrome in adults.
The objective was to develop evidence-based recommendations for the management of antiphospholipid syndrome (APS) in adults. Based on evidence from a systematic literature review and expert opinion, overarching principles and recommendations were formulated and voted. High-risk antiphospholipid antibody (aPL) profile is associated with greater risk for thrombotic and obstetric APS. Risk modification includes screening for and management of cardiovascular and venous thrombosis risk factors, patient education about treatment adherence, and lifestyle counselling. Low-dose aspirin (LDA) is recommended for asymptomatic aPL carriers, patients with systemic lupus erythematosus without prior thrombotic or obstetric APS, and non-pregnant women with a history of obstetric APS only, all with high-risk aPL profiles. Patients with APS and first unprovoked venous thrombosis should receive long-term treatment with vitamin K antagonists (VKA) with a target international normalised ratio (INR) of 2-3. In patients with APS with first arterial thrombosis, treatment with VKA with INR 2-3 or INR 3-4 is recommended, considering the individual's bleeding/thrombosis risk. Rivaroxaban should not be used in patients with APS with triple aPL positivity. For patients with recurrent arterial or venous thrombosis despite adequate treatment, addition of LDA, increase of INR target to 3-4 or switch to low molecular weight heparin may be considered. In women with prior obstetric APS, combination treatment with LDA and prophylactic dosage heparin during pregnancy is recommended. In patients with recurrent pregnancy complications, increase of heparin to therapeutic dose, addition of hydroxychloroquine or addition of low-dose prednisolone in the first trimester may be considered. These recommendations aim to guide treatment in adults with APS. High-quality evidence is limited, indicating a need for more research.


Lansoprazole for persistent throat symptoms in secondary care: the TOPPITS RCT.
Persistent throat symptoms are commonly attributed to 'laryngopharyngeal reflux'. Despite a limited evidence base, these symptoms are increasingly being treated in primary care with proton pump inhibitors. To assess the value of proton pump inhibitor therapy in patients with persistent throat symptoms. This was a double-blind, placebo-controlled, randomised Phase III trial. This was a multicentre UK trial in eight UK ear, nose and throat departments. A total of 346 participants aged ≥ 18 years with persistent throat symptoms and a Reflux Symptom Index score of ≥ 10, exclusive of the dyspepsia item, were recruited. Random allocation (1 : 1 ratio) to either 30 mg of lansoprazole twice daily or matched placebo for 16 weeks. Symptomatic response (i.e. total Reflux Symptom Index score after 16 weeks of therapy). A total of 1427 patients were screened and 346 were randomised. The mean age was 52 years (standard deviation 13.7 years, range 20-84 years); 150 (43%) participants were male and 196 (57%) were female; 184 (53%) participants had a mild Reflux Symptom Index minus the heartburn/dyspepsia item and 162 (47%) had a severe Reflux Symptom Index minus the heartburn/dyspepsia item. A total of 172 patients were randomised to lansoprazole and 174 were randomised to placebo. A total of 267 participants completed the primary end-point visit (lansoprazole, n = 127; placebo, n = 140), of whom 220 did so between 14 and 20 weeks post randomisation ('compliant' group); 102 received lansoprazole and 118 received placebo. The mean Reflux Symptom Index scores at baseline were similar [lansoprazole 22.0 (standard deviation 8.0), placebo 21.7 (standard deviation 7.1), overall 21.9 (standard deviation 7.5)]. The mean Reflux Symptom Index scores at 16 weeks reduced from baseline in both groups [overall 17.4 (standard deviation 9.9), lansoprazole 17.4 (standard deviation 9.9), placebo 15.6 (standard deviation 9.8)]. Lansoprazole participants had estimated Reflux Symptom Index scores at 16 weeks that were 1.9 points higher (worse) than those of placebo participants (95% confidence interval -0.3 to 4.2; padj = 0.096), adjusted for site and baseline severity. Ninety-five (43%) participants achieved a Reflux Symptom Index score in the normal range (< 12) at 16 weeks: 42 (41%) in the lansoprazole group and 53 (45%) in the placebo group. A total of 226 participants completed the end-of-trial follow-up visit (lansoprazole, n = 109; placebo, n = 117), of whom 181 were 'compliant'. The mean Reflux Symptom Index scores at 12 months reduced from baseline in both groups [lansoprazole 16.0 (standard deviation 10.8), placebo 13.6 (standard deviation 9.6), overall 14.7 (standard deviation 10.2)]. A total of 87 (48%) participants achieved a Reflux Symptom Index score in the normal range at 12 months: 33 (40%) in the lansoprazole group and 54 (55%) in the placebo group. Likewise, the Comprehensive Reflux Symptom Score and Laryngopharyngeal Reflux - Health Related Quality of Life total scores and subscales all showed very similar changes in the lansoprazole and placebo cohorts at both 16 weeks and 12 months. Drop-out rate and compliance are issues in pragmatic clinical trials. The Trial Of Proton Pump Inhibitors in Throat Symptoms (TOPPITS) aimed to detect clinically relevant difference with 90% power. The 346 randomised participants reduced to 283 at the primary end point; 267 completed the primary outcome measure, 220 within the protocol time scale. Despite this, the powers to detect the clinically relevant difference in Reflux Symptom Index score at 16 weeks were 82% (compliant comparison) and 89% (pragmatic comparison). The lack of difference between lansoprazole and placebo is generalisable across NHS clinics. Participants on lansoprazole did not report significantly better outcomes than participants on placebo on any of the three patient-reported outcome tools (Reflux Symptom Index, Comprehensive Reflux Symptom Score and Laryngopharyngeal Reflux - Health Related Quality of Life). This multicentre, pragmatic, powered, definitive Phase III trial found no evidence of benefit for patients by treating persistent throat symptoms with lansoprazole. Current Controlled Trials ISRCTN38578686 and EudraCT number 2013-004249-17. This project was funded by the National Institute for Health Research (NIHR) Health Technology Assessment programme and will be published in full in Health Technology Assessment; Vol. 25, No. 3. See the NIHR Journals Library website for further project information.


The experiences of children with intellectual and developmental disabilities in inclusive schools in Accra, Ghana.
Inclusive education is internationally recognised as the best strategy for providing equitable quality education to all children. However, because of the unique challenges they often present, children with intellectual and developmental disabilities (IDDs) are often excluded from inclusive schools. To date, limited research on inclusion has been conducted involving children with IDD as active participants. The study sought to understand the experiences of children with IDDs in learning in inclusive schools in Accra, Ghana. A qualitative descriptive design was utilised with 16 children with IDDs enrolled in inclusive schools in Accra, Ghana. Participants were recruited through purposive sampling and data were collected using classroom observations, the draw-and-write technique and semi-structured interviews. The data were analysed to identify themes as they emerged. Children's experiences in inclusive schools were identified along three major themes: (1) individual characteristics, (2) immediate environments and (3) interactional patterns. Insights from children's experiences reveal that they faced challenges including corporal punishment for slow performance, victimisation and low family support relating to their learning. Although children with IDDs receive peer support in inclusion, they experience diverse challenges including peer victimisation, corporal punishment and low family and teacher support in their learning. Improvement in inclusive best practices for children with IDD requires systematic efforts by diverse stakeholders to address identified challenges.


Small ruminant lentiviruses (SRLVs) break the species barrier to acquire new host range.
Zoonotic events of simian immunodeficiency virus (SIV) from non-human primates to humans have generated the acquired immunodeficiency syndrome (AIDS), one of the most devastating infectious disease of the last century with more than 30 million people dead and about 40.3 million people currently infected worldwide. Human immunodeficiency virus (HIV-1 and HIV-2), the two major viruses that cause AIDS in humans are retroviruses of the lentivirus genus. The genus includes arthritis-encephalitis virus (CAEV) and Maedi-Visna virus (MVV), and a heterogeneous group of viruses known as small ruminant lentiviruses (SRLVs), affecting goat and sheep. Lentivirus genome integrates into the host DNA, causing persistent infection associated with a remarkable diversity during viral replication. Direct evidence of mixed infections with these two closely related SRLVs was found in both sheep and goats. The evidence of a genetic continuum with caprine and ovine field isolates demonstrates the absence of an efficient species barrier preventing cross-species transmission. In dual-infected animals, persistent infections with both CAEV and MVV have been described, and viral chimeras have been detected. This not only complicates animal trade between countries but favors the risk that highly pathogenic variants may emerge as has already been observed in the past in Iceland and, more recently, in outbreaks with virulent strains in Spain. SRLVs affecting wildlife have already been identified, demonstrating the existence of emergent viruses adapted to new hosts. Viruses adapted to wildlife ruminants may acquire novel biopathological properties which may endanger not only the new host species but also domestic ruminants and humans. SRLVs infecting sheep and goats follow a genomic evolution similar to that observed in HIV or in other lentiviruses. Lentivirus genetic diversity and host factors leading to the establishment of naturally occurring virulent versus avirulent infections, in addition to the emergence of new strains, challenge every aspect of SRLV control measures for providing efficient tools to prevent the transmission of diseases between wild ungulates and livestock.


Mechanisms of Connexin-Related Lymphedema.
Mutations in GJC2 and GJA1, encoding Cxs (connexins) 47 and 43, respectively, are linked to lymphedema, but the underlying mechanisms are unknown. Because efficient lymph transport relies on the coordinated contractions of lymphatic muscle cells (LMCs) and their electrical coupling through Cxs, Cx-related lymphedema is proposed to result from dyssynchronous contractions of lymphatic vessels. To determine which Cx isoforms in LMCs and lymphatic endothelial cells are required for the entrainment of lymphatic contraction waves and efficient lymph transport. We developed novel methods to quantify the spatiotemporal entrainment of lymphatic contraction waves and used optogenetic techniques to analyze calcium signaling within and between the LMC and the lymphatic endothelial cell layers. Genetic deletion of the major lymphatic endothelial cell Cxs (Cx43, Cx47, or Cx37) revealed that none were necessary for the synchronization of the global calcium events that triggered propagating contraction waves. We identified Cx45 in human and mouse LMCs as the critical Cx mediating the conduction of pacemaking signals and entrained contractions. Smooth muscle-specific Cx45 deficiency resulted in 10- to 18-fold reduction in conduction speed, partial-to-severe loss of contractile coordination, and impaired lymph pump function ex vivo and in vivo. Cx45 deficiency resulted in profound inhibition of lymph transport in vivo, but only under an imposed gravitational load. Our results (1) identify Cx45 as the Cx isoform mediating the entrainment of the contraction waves in LMCs; (2) show that major endothelial Cxs are dispensable for the entrainment of contractions; (3) reveal a lack of coupling between lymphatic endothelial cells and LMCs, in contrast to arterioles; (4) point to lymphatic valve defects, rather than contraction dyssynchrony, as the mechanism underlying GJC2- or GJA1-related lymphedema; and (5) show that a gravitational load exacerbates lymphatic contractile defects in the intact mouse hindlimb, which is likely critical for the development of lymphedema in the adult mouse.


Identification of UNC5A as a novel transcriptional target of tumor suppressor p53 and a regulator of apoptosis.
UNC5A is an axon-guidance molecule, and plays a critical role in neuronal development and differentiation as a netrin-1 receptor. Emerging evidence suggests that axon guidance molecules including UNC5A regulate apoptosis in non-neuronal cells. Here, we report that UNC5A regulates apoptosis as a downstream target of p53. UNC5A expression was strongly induced by exogenous and endogenous p53. Chromatin immunoprecipitation (ChIP) revealed that p53 binds to a sequence in the promoter region of the UNC5A gene. Reporter assays showed that this sequence exhibits p53-dependent transcriptional activity. Overexpression of UNC5A significantly suppressed colony formation of two glioblastoma cell lines-U373MG and T98G. UNC5A dramatically induced apoptosis through the activation of caspase-3 in various cancer cell lines, including LS174T (colon cancer), U373MG (glioblastoma), SH-SY5Y (neuroblastoma), and SKNAS (neuroblastoma). Finally, gamma irradiation strongly induced the expression of UNC5A mRNA in the spleen and colon of p53+/+ mice, but not in those of p53-/- mice, implying that the transcription of UNC5A in vivo is regulated by p53. These results suggest that UNC5A is a novel transcriptional target of p53 and plays a role in p53-dependent apoptosis.


Integration of Proteomics and Transcriptomics Data Sets for the Analysis of a Lymphoma B-Cell Line in the Context of the Chromosome-Centric Human Proteome Project.
A comprehensive study of the molecular active landscape of human cells can be undertaken to integrate two different but complementary perspectives: transcriptomics, and proteomics. After the genome era, proteomics has emerged as a powerful tool to simultaneously identify and characterize the compendium of thousands of different proteins active in a cell. Thus, the Chromosome-centric Human Proteome Project (C-HPP) is promoting a full characterization of the human proteome combining high-throughput proteomics with the data derived from genome-wide expression profiling of protein-coding genes. Here we present a full proteomic profiling of a human lymphoma B-cell line (Ramos) performed using a nanoUPLC-LTQ-Orbitrap Velos proteomic platform, combined to an in-depth transcriptomic profiling of the same cell type. Data are available via ProteomeXchange with identifier PXD001933. Integration of the proteomic and transcriptomic data sets revealed a 94% overlap in the proteins identified by both -omics approaches. Moreover, functional enrichment analysis of the proteomic profiles showed an enrichment of several functions directly related to the biological and morphological characteristics of B-cells. In turn, about 30% of all protein-coding genes present in the whole human genome were identified as being expressed by the Ramos cells (stable average of 30% genes along all the chromosomes), revealing the size of the protein expression-set present in one specific human cell type. Additionally, the identification of missing proteins in our data sets has been reported, highlighting the power of the approach. Also, a comparison between neXtProt and UniProt database searches has been performed. In summary, our transcriptomic and proteomic experimental profiling provided a high coverage report of the expressed proteome from a human lymphoma B-cell type with a clear insight into the biological processes that characterized these cells. In this way, we demonstrated the usefulness of combining -omics for a comprehensive characterization of specific biological systems.


Renal alpha 1-adrenergic receptor subtypes: MDCK-D1 cells, but not rat cortical membranes possess a single population of receptors.
Recent work has demonstrated that alpha 1-adrenergic receptors are composed of at least two subtypes, termed alpha 1a and alpha 1b. It has been proposed that these subtypes may be linked to distinct second messenger systems. In the current studies, we have compared the properties of alpha 1-adrenergic receptors in rat renal cortical membranes with those in MDCK-D1 cells, a clonal cell line derived from distal tubule/collecting duct. Competitive binding studies with [3H]prazosin and compounds [5-methylurapidil, (+)-niguldipine, WB4101, and oxymetazoline] that distinguish high affinity (alpha 1a) and low affinity (alpha 1b) sites indicated that rat renal cortical membranes contain about 50% of each class of site. In contrast, MDCK-D1 cells contained a single population of low affinity sites. 5-Methylurapidil, but not the other compounds, recognized binding sites in these cells with a substantially lower affinity than has been observed for the low affinity site in other tissues and in parallel studies with renal cortical membranes. [3H]Prazosin binding sites in these cells, as well as alpha 1-adrenergic receptor-mediated arachidonic acid release and phosphoinositide and phosphatidylcholine hydrolysis, were sensitive to inactivation by chloroethylclonidine (IC50 approximately 0.7 microM), as expected for alpha 1b receptors. However, alpha 1-adrenergic receptors of MDCK-D1 cells required extracellular calcium for biological response, unlike what has been hypothesized for the alpha 1b receptor subtype. These data indicate that the population of alpha 1-adrenergic receptors of distal tubule/collecting duct cells likely consists of receptors of the alpha 1b subtype. The low affinity binding of 5-methylurapidil and the requirement for extracellular calcium for biological response in these cells suggest that this receptor may not be identical to the alpha 1b receptor that has been observed in other systems.


Double strand breaks can initiate gene silencing and SIRT1-dependent onset of DNA methylation in an exogenous promoter CpG island.
Chronic exposure to inducers of DNA base oxidation and single and double strand breaks contribute to tumorigenesis. In addition to the genetic changes caused by this DNA damage, such tumors often contain epigenetically silenced genes with aberrant promoter region CpG island DNA hypermethylation. We herein explore the relationships between such DNA damage and epigenetic gene silencing using an experimental model in which we induce a defined double strand break in an exogenous promoter construct of the E-cadherin CpG island, which is frequently aberrantly DNA hypermethylated in epithelial cancers. Following the onset of repair of the break, we observe recruitment to the site of damage of key proteins involved in establishing and maintaining transcriptional repression, namely SIRT1, EZH2, DNMT1, and DNMT3B, and the appearance of the silencing histone modifications, hypoacetyl H4K16, H3K9me2 and me3, and H3K27me3. Although in most cells selected after the break, DNA repair occurs faithfully with preservation of activity of the promoter, a small percentage of the plated cells demonstrate induction of heritable silencing. The chromatin around the break site in such a silent clone is enriched for most of the above silent chromatin proteins and histone marks, and the region harbors the appearance of increasing DNA methylation in the CpG island of the promoter. During the acute break, SIRT1 appears to be required for the transient recruitment of DNMT3B and subsequent methylation of the promoter in the silent clones. Taken together, our data suggest that normal repair of a DNA break can occasionally cause heritable silencing of a CpG island-containing promoter by recruitment of proteins involved in silencing. Furthermore, with contribution of the stress-related protein SIRT1, the break can lead to the onset of aberrant CpG island DNA methylation, which is frequently associated with tight gene silencing in cancer.


Adenovirus with hexon Tat-protein transduction domain modification exhibits increased therapeutic effect in experimental neuroblastoma and neuroendocrine tumors.
Adenovirus serotype 5 (Ad5) is widely used as an oncolytic agent for cancer therapy. However, its infectivity is highly dependent on the expression level of coxsackievirus-adenovirus receptor (CAR) on the surfaces of tumor cells. Furthermore, infected cells overproduce adenovirus fiber proteins, which are released prior to cell lysis. The released fibers block CAR on noninfected neighboring cells, thereby preventing progeny virus entry. Our aim was to add a CAR-independent infection route to Ad5 to increase the infectivity of tumor cells with low CAR expression and prevent the fiber-masking problem. We constructed Ad5 viruses that encode the protein transduction domain (PTD) of the HIV-1 Tat protein (Tat-PTD) in hypervariable region 5 (HVR5) of the hexon protein. Tat-PTD functions as a cell-penetrating peptide, and Tat-PTD-modified Ad5 showed a dramatic increased transduction of CAR-negative cell lines compared to unmodified vector. Moreover, while tumor cell infectivity was severely reduced for Ad5 in the presence of fiber proteins, it was only marginally reduced for Tat-PTD-modified Ad5. Furthermore, because of the sequence alteration in the hexon HVR, coagulation factor X-mediated virus uptake was significantly reduced. Mice harboring human neuroblastoma and neuroendocrine tumors show suppressed tumor growths and prolonged survival when treated with Tat-PTD-modified oncolytic viruses. Our data suggest that modification of Ad5 with Tat-PTD in HVR5 expands its utility as an oncolytic agent.


The Temporal and Spatial Invasion Genetics of the Western Corn Rootworm (Coleoptera: Chrysomelidae) in Southern Europe.
This study describes the genetics of the western corn rootworm (WCR), Diabrotica virgifera virgifera LeConte in southern Europe during the introduction (1996-2001) and establishment/spread (2002-2011) phases of its invasion. The Diabrotica microsatellite core-set was used to perform traditional population genetics analyses. Our results indicated that during the introduction phase genetic diversity and population genetic structure were lower overall as compared to the establishment/spread phase. Unusually high genetic differentiation was found between the Italy and southern Europe comparisons, including high differentiation between Italian populations separated by a short distance during the establishment/spread phase. STRUCTURE analysis revealed two genetic clusters during the introduction phase and two genetic clusters during the establishment/spread phase. However, bottlenecked populations were only detected during the invasion phase. A small but significant isolation by distance effect was noted in both phases. Serbia was the geographic source of WCR to Croatia and Hungary in the introduction phase, while the United States of America was the possible source of WCR to Italy in 2001. These introductory populations were the subsequent source of individuals sampled during the establishment/spread phase. Repeated introductions and admixture events in southern Europe may have resulted in genetically diverse WCR populations that have attained 83% of all known alleles worldwide.


Genetic influences on language impairment and phonological short-term memory.
It has been known for some years that specific language impairment (SLI), an unexpected failure to acquire age-appropriate language skills, is highly heritable. However, molecular genetic studies have been hampered by the heterogeneity of the disorder and the predominant lack of clear genotype-phenotype relationships. We review recent studies suggesting that a better understanding of the genetics of SLI might emerge if we move away from clinical criteria for diagnosis to look instead at a theoretically based quantitative and cognitive measure of the phenotype: a test of phonological short-term memory (STM). Deficient phonological STM has been linked to specific genetic loci, and might play a role in determining some types of reading impairment as well as SLI. Identifying those cognitive deficits that work best as indices of heritable phenotypes will help us to uncover the aetiology of developmental disorders.


The impact of quality and accessibility of primary care on emergency admissions for a range of chronic ambulatory care sensitive conditions (ACSCs) in Scotland: longitudinal analysis.
Hospital admissions for Ambulatory Care Sensitive Conditions (ACSC) are those that could potentially be prevented by timely and effective disease management within primary care. ACSC admissions are increasingly used as performance indicators. However, key questions remain about the validity of these measures. The evidence to date has been inconclusive and limited to specific conditions. The aim of this study was to test the robustness of ACSC admissions as indicators of the quality of primary care. It is the first study to examine a wide range of ACSCs using longitudinal data which enables us to control for unmeasured characteristics which differ by practice but which are constant over time. Using longitudinal data at the practice level, from 907 Scottish practices for the time period 1/4/2005 to 31/32012, we explored the relationships between the quality of primary care, and hospital admissions for multiple ACSCs controlling for a wide range of covariates including characteristics of GP practices, characteristics of the practice population, hospital effects and year effects. We examined the impact of two dimensions of quality of care: clinical quality of and access to daytime general practice. Generalised Estimating Equations taking the form of Negative Binomial regression models with the practice population included as the exposure term were estimated. We found that higher achievement on some clinical quality measures of primary care was associated with reduced ACSC emergency admissions. We also show that access to primary care was associated with ACSC emergency admissions. However, the effects were small and inconsistent and ACSC emergency admissions were associated with several confounding factors such as deprivation, rurality and distance to the hospital. The results suggest caution in the use of crude ACSC admission rates as a performance indicator of quality of primary care.


Late oligodendrocyte progenitors coincide with the developmental window of vulnerability for human perinatal white matter injury.
Hypoxic-ischemic injury to the periventricular cerebral white matter [periventricular leukomalacia (PVL)] results in cerebral palsy and is the leading cause of brain injury in premature infants. The principal feature of PVL is a chronic disturbance of myelination and suggests that oligodendrocyte (OL) lineage progression is disrupted by ischemic injury. We determined the OL lineage stages at risk for injury during the developmental window of vulnerability for PVL (23-32 weeks, postconceptional age). In 26 normal control autopsy human brains, OL lineage progression was defined in parietal white matter, a region of predilection for PVL. Three successive OL stages, the late OL progenitor, the immature OL, and the mature OL, were characterized between 18 and 41 weeks with anti-NG2 proteoglycan, O4, O1, and anti-myelin basic protein (anti-MBP) antibodies. NG2+O4+ late OL progenitors were the predominant stage throughout the latter half of gestation. Between 18 and 27 weeks, O4+O1+ immature OLs were a minor population (9.9 +/- 2.1% of total OLs; n = 9). Between 28 and 41 weeks, an increase in immature OLs to 30.9 +/- 2.1% of total OLs (n = 9) was accompanied by a progressive increase in MBP+ myelin sheaths that were restricted to the periventricular white matter. The developmental window of high risk for PVL thus precedes the onset of myelination and identifies the late OL progenitor as the major potential target. Moreover, the decline in incidence of PVL at approximately 32 weeks coincides with the onset of myelination in the periventricular white matter and suggests that the risk for PVL is related to the presence of late OL progenitors in the periventricular white matter.


Adaptive inferential sensors based on evolving fuzzy models.
A new technique to the design and use of inferential sensors in the process industry is proposed in this paper, which is based on the recently introduced concept of evolving fuzzy models (EFMs). They address the challenge that the modern process industry faces today, namely, to develop such adaptive and self-calibrating online inferential sensors that reduce the maintenance costs while keeping the high precision and interpretability/transparency. The proposed new methodology makes possible inferential sensors to recalibrate automatically, which reduces significantly the life-cycle efforts for their maintenance. This is achieved by the adaptive and flexible open-structure EFM used. The novelty of this paper lies in the following: (1) the overall concept of inferential sensors with evolving and self-developing structure from the data streams; (2) the new methodology for online automatic selection of input variables that are most relevant for the prediction; (3) the technique to detect automatically a shift in the data pattern using the age of the clusters (and fuzzy rules); (4) the online standardization technique used by the learning procedure of the evolving model; and (5) the application of this innovative approach to several real-life industrial processes from the chemical industry (evolving inferential sensors, namely, eSensors, were used for predicting the chemical properties of different products in The Dow Chemical Company, Freeport, TX). It should be noted, however, that the methodology and conclusions of this paper are valid for the broader area of chemical and process industries in general. The results demonstrate that well-interpretable and with-simple-structure inferential sensors can automatically be designed from the data stream in real time, which predict various process variables of interest. The proposed approach can be used as a basis for the development of a new generation of adaptive and evolving inferential sensors that can address the challenges of the modern advanced process industry.


A clinical study on the 6-year outcomes of immediately loaded three implants for completely edentulous mandibles: "the all-on-3 concept".
The objective of the current study was to demonstrate the 6-year clinical and radiological treatment outcomes of the technique performed by immediately loading of three implants (single straight in the mid-line and two tilted distal implants) for the management of total edentulous mandibles and introduce a simple decision matrix for selection of the most appropriate protocol in cases with insufficient length of the interforaminal area. Re-assessments were performed over a total observation period of 6 years after surgery via measurement of bone resorption around implants, bleeding on probing, plaque accumulation, periodontal probing depth, bite force measurements and oral health impact profile. A total of 29 patients (45% women and 55% men) with a mean age of 65 ± 6 years enrolled in the study. 14 patients received an acryl-based bridge as definitive prosthetic restoration and 15 patients received a ceramic-based restoration. Both during the immediate loading phase and during the 6-year follow-up, there was no implantation loss. Regardless of the implant position, all implants showed continuous bone loss over the observation time. The bone loss around dental implants during observation period was only maximum 1.0 ± 1 mm and it remained well within the limits for 'success' according to the 2007 Pisa consensus (< 2 mm). The plaque index showed no significant fluctuations between the implant positions and the individual examination times. The approach described herein might help the surgeon by avoiding unnecessary loss of bone strength, selecting implant sites, and establishing the biomechanical advantage of increased A-P spread for immediate function.


Inhibition of the alveolar macrophage oxidative burst by a diffusible component from the surface of the spores of the fungus Aspergillus fumigatus.
Aspergillus fumigatus is a fungus that grows on dead and decaying organic matter in the environment and whose spores are present ubiquitously in the air. The fungus causes a range of diseases in the human lung. A study was undertaken to demonstrate and partially characterise an inhibitor of the macrophage respiratory burst from the surface of A fumigatus spores that could be an important factor in allowing the fungus to colonise the lung. The spore-derived inhibitor of the respiratory burst of rat alveolar macrophages, as measured by generation of superoxide anion, was demonstrated in Hank's balanced salt solution extracts of four clinical isolates and an environmental isolate of A fumigatus. The time course of the release of the inhibitor into aqueous solution was assessed and the cytotoxic potential of the spore-derived inhibitor towards macrophages was tested using the propidium iodide method. An oxygen electrode was used to confirm the superoxide anion measurements. Molecular weight cutoff filters were used to determine the size of the inhibitor as assessed in the respiratory burst assay and also by its ability to inhibit macrophage spreading on glass. The crude diffusate from the spore surface was fractionated by reversed phase high pressure liquid chromatography (HPLC) and the fractions analysed for inhibitory activity, protein, and carbohydrate content. A small molecular weight (< 10 kD) heat stable toxin was released from the spores of clinical and environmental isolates of A fumigatus within minutes of deposition in aqueous solution. The key effect of the toxin demonstrated here was its ability to inhibit the oxidative burst of macrophages as measured by superoxide anion release. The inhibition was not due to cell death or detectable loss of membrane integrity as measured by permeability to propidium iodide. The toxin was not a scavenger of superoxide anion. Oxygen electrode studies suggested indirectly that the inhibitor acted to inhibit the assembly of the macrophage NADPH-oxidase complex. Fractions of < 10 kD also inhibited the spreading of alveolar macrophages, confirming that the toxin had an additional effect on macrophages that leads to loss of adherence or impairment of cytoskeletal function. In reversed phase HPLC fractions the inhibitory activity eluted with an associated carbohydrate, although the exact chemical nature of the toxin remains to be elucidated. This spore toxin may, through its ability to diffuse rapidly into lung lining fluid, diminish the macrophage respiratory burst and play a part in allowing A fumigatus to persist in the lung and manifest its well known pathogenic effects. Future research will be focused on further molecular characterisation of the toxin and elaboration of the effect of the toxin on intracellular signalling pathways involved in the activation of alveolar macrophages.


Review of cases of IgA nephropathy.
A review of 26 years of British military renal pathology showed the commonest diagnosis to be immunoglobulin A (IgA) nephropathy affecting 115/346 (33%) of cases. It was possible to follow-up 50/115 military patients with this condition with the primary objective to determine whether initial observations enabled a confident prediction of prognosis. Additionally, unpublished observations have shown that the incidence of glomerulonephritis in an Indo-Asian British military racial group was fourfold that in the majority Caucasians although the nature of pathology (IgA vs non-IgA nephropathy) was not statistically significantly different. The present study secondarily sought to determine if prognosis of IgA nephropathy was different in this Indo-Asian military group. Finally, some conclusions concerning the traditionally restrictive policy towards applicants for military service with evidence of active nephritis were attempted. An archive of military renal patients covering 1985-2011 was reviewed and clinical details of cases of IgA nephropathy were extracted and analysed. 95 cases (80 Caucasian, 15 Indo-Asian) were reviewed. There was no racial difference (p=0.2) in initial median estimated glomerular filtration rate (eGFR, 86 vs 83 ml/min/1.73 m(2)). Altogether, initial median of mean arterial pressure (MAP) was 96 mm Hg (IQR=87-103) and median proteinuria was 485 mg/24 h (IQR=195-925). There was an inverse correlation between initial eGFR and both MAP (p=0.0008) and proteinuria (p=0.0006). In the 50 patients who were followed up, the change in eGFR with time (ΔeGFR) was calculated. 10 of the 44 Caucasians and 3/6 Indo-Asians showed significant changes in eGFR but population proportions were not significantly different (p=0.2). Altogether, 11/50 (22%) showed deteriorating eGFR despite therapeutic interventions, compared with those without deterioration; this was not due to different duration of observation (p=0.08), initial MAP (p=0.23) or initial proteinuria (p=0.07). There was no statistically significant correlation between ΔeGFR and initial MAP (p=0.6) or proteinuria (p=0.7). The proportion of military cases of IgA nephropathy with deteriorating renal function (22%) was not different from that described in civilians, suggesting that military medical management (including operational restriction where necessary) was appropriate. Initial clinical observations, including racial group, did not permit confident prediction of prognosis thus mandating follow-up for all military cases. Relaxing current enlistment policy would be inadvisable given the inability to confidently predict prognosis in individual cases.


Accreditation standards for medical imaging services.
The rapid changes in the healthcare system, with revolutionary advancements in imaging, along with the lack of any existing imaging standards in our country, have raised the need for an accreditation structure. The Quality Council of India (QCI) has therefore introduced standards for medical imaging services, focusing on the control of services, personnel, imaging processes and procedures, facility and environment, equipment, and documentation, as well as risk control and safety. This article deals briefly with the standards structured by the QCI for accreditation of imaging services.


Scaling up health knowledge at European level requires sharing integrated data: an approach for collection of database specification.
Computerized health care databases have been widely described as an excellent opportunity for research. The availability of "big data" has brought about a wave of innovation in projects when conducting health services research. Most of the available secondary data sources are restricted to the geographical scope of a given country and present heterogeneous structure and content. Under the umbrella of the European Innovation Partnership on Active and Healthy Ageing, collaborative work conducted by the partners of the group on "adherence to prescription and medical plans" identified the use of observational and large-population databases to monitor medication-taking behavior in the elderly. This article describes the methodology used to gather the information from available databases among the Adherence Action Group partners with the aim of improving data sharing on a European level. A total of six databases belonging to three different European countries (Spain, Republic of Ireland, and Italy) were included in the analysis. Preliminary results suggest that there are some similarities. However, these results should be applied in different contexts and European countries, supporting the idea that large European studies should be designed in order to get the most of already available databases.


Evaluation of antiurolithiatic and antioxidant potential of Lepidagathis prostrata: A Pashanbhed plant.
Oxidative stress acts as an essential mediator in the pathophysiology of urolithiasis. Lepidagathis prostrata Dalz. (Acanthaceae) is a Pashanbhed plant that is recommended for the management of urolithiasis; however, no scientific validation has been reported. To evaluate the antiurolithiatic and antioxidant potential of L. prostrata. Methanol extract (LPM) and fractions; petroleum ether (LPPE), ethyl acetate (LPEA), n-butanol (LPBU) and aqueous (LPAQ) were prepared. In vitro antiurolithiatic activity was evaluated by the capacity to inhibit calcium oxalate (CaOx) nucleation and aggregation at different concentrations of extract/fractions (0.04-3 mg/mL) for 30 min. Total phenol and flavonoid content and antioxidant potential were determined. A validated HPTLC method was performed to quantify lupeol and β-sitosterol. LPEA exhibited the highest dose-dependent inhibition of CaOx nucleation (IC50: 336.23 ± 30.79 µg/mL) and aggregation (IC50: 149.63 ± 10.31 µg/mL), which was significantly (p < 0.05) better than standard Cystone®. The polar LPBU fraction was enriched with phenols (47.34 ± 0.19 mg GAE/g) and flavonoids (20.38 ± 0.05 mg QE/g), which correlates with its highest antioxidant potential in DPPH, ABTS, nitric oxide scavenging and iron chelating activities (IC50: 1.18-87.34 µg/mL). To our knowledge, this is the first study reporting the presence of lupeol and β-sitosterol in L. prostrata. The antiurolithiatic activity of L. prostrata is probably mediated through the inhibition of CaOx crystallization. In addition to its free radical scavenging and antioxidant activities, it would act as an excellent agent for the prevention of urolithiasis.


Inbreeding Ratio and Genetic Relationships among Strains of the Western Clawed Frog, Xenopus tropicalis.
The Western clawed frog, Xenopus tropicalis, is a highly promising model amphibian, especially in developmental and physiological research, and as a tool for understanding disease. It was originally found in the West African rainforest belt, and was introduced to the research community in the 1990s. The major strains thus far known include the Nigerian and Ivory Coast strains. However, due to its short history as an experimental animal, the genetic relationship among the various strains has not yet been clarified, and establishment of inbred strains has not yet been achieved. Since 2003 the Institute for Amphibian Biology (IAB), Hiroshima University has maintained stocks of multiple X. tropicalis strains and conducted consecutive breeding as part of the National BioResource Project. In the present study we investigated the inbreeding ratio and genetic relationship of four inbred strains at IAB, as well as stocks from other institutions, using highly polymorphic microsatellite markers and mitochondrial haplotypes. Our results show successive reduction of heterozygosity in the genome of the IAB inbred strains. The Ivory Coast strains clearly differed from the Nigerian strains genetically, and three subgroups were identified within both the Nigerian and Ivory Coast strains. It is noteworthy that the Ivory Coast strains have an evolutionary divergent genetic background. Our results serve as a guide for the most effective use of X. tropicalis strains, and the long-term maintenance of multiple strains will contribute to further research efforts.


Relative Distribution, Diversity, and Bloodmeal Sources of Mosquitoes and Known Vectors of Rift Valley Fever Phlebovirus in Three Differing Ecosystems in Bura, Tana River County, Kenya.
Environmental modifications disturb the equilibrium of mosquito populations, altering the risk of mosquito-borne diseases. Mosquito distribution, diversity, and bloodmeal sources were examined to compare Rift Valley fever (RVF) risk among irrigated, riverine, and pastoral ecosystems in Bura, Tana River County, Kenya, between September 2014 and June 2015. Thirty-eight households and 21 irrigation fields were selected for the study. Mosquitoes were trapped with carbon dioxide-impregnated CDC traps, one trap per household and three traps per irrigated field, and morphologically identified using taxonomic keys. Host DNA was extracted from engorged females and cytochrome b genes amplified by PCR to identify sources of bloodmeals. A total of 21,015 mosquitoes were collected; 5742 within households in the 3 ecosystems and 15,273 within irrigated fields. Mosquitoes collected within irrigated fields belonged to 8 genera and 37 species, while those from households within the irrigation scheme belonged to 6 genera and 29 species. Collections from riverine and pastoral households belonged to five and four genera, respectively. The most abundant genera in the irrigated fields were Aedes (21%) and Mansonia (22%), while Anopheles (43%) was the most abundant within households. Most mosquitoes in riverine and pastoral households belonged to Anopheles (76%) and Aedes (65%) genera, respectively. Seasonal variation driven by rainfall was evidenced by spikes in mosquito numbers within irrigated and riverine ecosystems. Host species identification revealed that goats and humans were the main sources of bloodmeal. There was an overall increase in mosquito abundance and diversity as a result of the presence of the irrigated ecosystem in this county, and an increased availability of highly RVF-susceptible hosts as a result of the establishment and concentration of residential areas, promoting potential vector-host contacts. These results highlight the impact of anthropogenic changes on mosquito ecology, potentially heightening the risk of transmission and maintenance of RVF in this region.


MRI-guided vacuum-assisted breast biopsy: experience of a single tertiary referral cancer centre and prospects for the future.
MRI-guided vacuum-assisted breast biopsy (VABB) is used for suspicious breast cancer (BC) lesions which are detectable only with MRI: because the high sensitivity but limited specificity of breast MRI it is a fundamental tool in breast imaging divisions. We analyse our experience of MRI-guided VABB and critically discuss the potentialities of diffusion-weighted imaging (DWI) and artificial intelligence (AI) in this matter. We retrospectively analysed a population of consecutive women underwent VABB at our tertiary referral BC centre from 01/2011 to 01/2019. Reference standard was histological diagnosis or at least 1-year negative follow-up. McNemar, Mann-Whitney and χ2 tests at 95% level of significance were used as statistical exams. 217 women (mean age = 52, 18-72 years) underwent MRI-guided VABB; 11 were excluded and 208 MRI-guided VABB lesions were performed: 34/208 invasive carcinomas, 32/208 DCIS, 8/208 LCIS, 3/208 high-risk lesions and 131/208 benign lesions were reported. Accuracy of MRI-guided VABB was 97%. The predictive features for malignancy were mass with irregular shape (OR 8.4; 95% CI 0.59-31.6), size of the lesion (OR 4.4; 95% CI 1.69-9.7) and mass with irregular/spiculated margins (OR 5.4; 95% CI 6.8-31.1). Six-month follow-up showed 4 false-negative cases (1.9%). Invasive BC showed a statistically significant higher hyperintense signal at DWI compared to benign lesions (p = 0.03). No major complications occurred. MR-guided VABB showed high accuracy. Benign-concordant lesions should be followed up with breast MRI in 6-12 months due to the risk of false-negative results. DWI and AI applications showed potential benefit as support tools for radiologists.


Breaking of macroscopic centric symmetry in paraelectric phases of ferroelectric materials and implications for flexoelectricity.
A centrosymmetric stress cannot induce a polar response in centric materials; piezoelectricity is, for example, possible only in non-centrosymmetric structures. An exception is metamaterials with shape asymmetry, which may be polarized by stress even when the material is centric. In this case the mechanism is flexoelectricity, which relates polarization to a strain gradient. The flexoelectric response scales inversely with size, thus a large effect is expected in nanoscale materials. Recent experiments in polycrystalline, centrosymmetric perovskites (for example, (Ba, Sr)TiO3) have indicated values of flexoelectric coefficients that are orders of magnitude higher than theoretically predicted, promising practical applications based on bulk materials. We show that materials with unexpectedly large flexoelectric response exhibit breaking of the macroscopic centric symmetry through inhomogeneity induced by the high-temperature processing. The emerging electro-mechanical coupling is significant and may help to resolve the controversy surrounding the large apparent flexoelectric coefficients in this class of materials.


Neurophysiological activity underlying altered brain metabolism in epileptic encephalopathies with CSWS.
We investigated the neurophysiological correlate of altered regional cerebral glucose metabolism observed in children with epileptic encephalopathy with continuous spike-waves during sleep (CSWS) by using a multimodal approach combining time-sensitive magnetic source imaging (MSI) and positron emission tomography with [(18)F]-fluorodeoxyglucose (FDG-PET). Six patients (4 boys and 2 girls, age range: 4-8 years, 3 patients with Landau-Kleffner syndrome (LKS), 3 patients with atypical rolandic epilepsy (ARE)) were investigated by FDG-PET and MSI at the acute phase of CSWS. In all patients, the onset(s) of spike-waves discharges were associated with significant focal hypermetabolism. The propagation of epileptic discharges to other brain areas was associated with focal hypermetabolism (five patients), hypometabolism (one patient) or the absence of any significant metabolic change (one patient). Interestingly, most of the hypometabolic areas were not involved in the epileptic network per se. This study shows that focal hypermetabolism observed at the acute phase of CSWS are related to the onset or propagation sites of spike-wave discharges. Spike-wave discharges propagation can be associated to other types of metabolic changes, suggesting the occurrence of various neurophysiological mechanisms at the cellular level. Most of the hypometabolic areas are not involved in the epileptic network as such and are probably related to a mechanism of remote inhibition. These findings highlight the critical value of combining FDG-PET with time-sensitive functional neuroimaging approaches such as MSI to assess CSWS epileptic network when surgery is considered as a therapeutic approach.


Physiologically based quantitative modeling of unihemispheric sleep.
Unihemispheric sleep has been observed in numerous species, including birds and aquatic mammals. While knowledge of its functional role has been improved in recent years, the physiological mechanisms that generate this behavior remain poorly understood. Here, unihemispheric sleep is simulated using a physiologically based quantitative model of the mammalian ascending arousal system. The model includes mutual inhibition between wake-promoting monoaminergic nuclei (MA) and sleep-promoting ventrolateral preoptic nuclei (VLPO), driven by circadian and homeostatic drives as well as cholinergic and orexinergic input to MA. The model is extended here to incorporate two distinct hemispheres and their interconnections. It is postulated that inhibitory connections between VLPO nuclei in opposite hemispheres are responsible for unihemispheric sleep, and it is shown that contralateral inhibitory connections promote unihemispheric sleep while ipsilateral inhibitory connections promote bihemispheric sleep. The frequency of alternating unihemispheric sleep bouts is chiefly determined by sleep homeostasis and its corresponding time constant. It is shown that the model reproduces dolphin sleep, and that the sleep regimes of humans, cetaceans, and fur seals, the latter both terrestrially and in a marine environment, require only modest changes in contralateral connection strength and homeostatic time constant. It is further demonstrated that fur seals can potentially switch between their terrestrial bihemispheric and aquatic unihemispheric sleep patterns by varying just the contralateral connection strength. These results provide experimentally testable predictions regarding the differences between species that sleep bihemispherically and unihemispherically.


Pathogenesis of oral type I feline infectious peritonitis virus (FIPV) infection: Antibody-dependent enhancement infection of cats with type I FIPV via the oral route.
Feline infectious peritonitis virus (FIPV) causes a severe, immune-mediated disease called FIP in domestic and wild cats. It is unclear whether FIP transmits from cat to cat through the oral route of FIPV infection, and the reason for this includes that FIP is caused by oral inoculation with some FIPV strains (e.g., type II FIPV WSU 79-1146), but is not caused by other FIPV (e.g., type I FIPV KU-2 strain: FIPV-I KU-2). In this study, when cats passively immunized with anti-FIPV-I KU-2 antibodies were orally inoculated with FIPV-I KU-2, FIP was caused at a 50% probability, i.e., FIPV not causing FIP through oral infection caused FIP by inducing antibody-dependent enhancement. Many strains of type I FIPV do not cause FIP by inoculation through the oral route in cats. Based on the findings of this study, type I FIPV which orally infected cats may cause FIP depending on the condition.


Quantitative study of the gray-scale fidelity of volume holographic images.
The reconstruction fidelity of gray-scale holographic images is analyzed quantitatively and assessed by use of proposed quantities, especially the correlation coefficient between the original and the retrieved images. The physical factors affecting the fidelity of photorefractive holograms are investigated experimentally, leading to results that are useful for the determination of the recording-beam ratio and the exposure dosage for high-fidelity retrieval. Investigation of the effects of noise on the fidelity of multiplexed holograms suggests that, if multiple-gray-scale data pages are multiplexed in a given volume, the total bit capacity may be competitive with that realized by means of storing binary pages.


Overview of Leafy Greens-Related Food Safety Incidents with a California Link: 1996 to 2016.
An increase in the number of foodborne illness outbreaks associated with produce has been noted in the literature, and leafy greens have been the most common produce category associated with these outbreaks. California is the largest leafy greens producer in the United States, and many related foodborne illness incidents were traced to this state. A systematic overview of leafy greens incidents linked to California was conducted by the California Department of Public Health, Food and Drug Branch through analysis of complaints, routine surveillance sampling, disease outbreaks, and investigations covering 1996 to 2016. The goal was to develop a risk assessment tool to modernize emergency response efforts to foodborne illnesses related to leafy greens. A database including environmental, epidemiologic, and laboratory information for each incident was developed, and descriptive analysis was performed to identify trends. In the 21-year period analyzed, 134 incidents were identified, the majority of which were surveillance related. Approximately 2,240 U.S. cases of confirmed illness were reported (298 California cases resulting in 50 hospitalizations). Romaine lettuce and spinach were the most commonly implicated vehicles. The most prevalent hazard type was microbiological, in particular bacterial, specifically associated with pathogenic strains of Escherichia coli. In California, the overview provided the Food and Drug Branch with a platform to (i) enhance its Food Safety Program, Emergency Response Unit, and California Food Emergency Response Team; (ii) assist in more efficient investigation, response, control, and prevention of California-linked foodborne illness incidents; and (iii) identify knowledge gaps and develop effective definitions, procedures, training, guidelines, and policies that will be used to help prevent future outbreaks. Outcomes provide insight into the situation in the largest leafy greens-producing state and may be used to prioritize limited national food safety resources and aid in future leafy greens-related research and foodborne incident investigations.


Relationships between hippocampal atrophy, white matter disruption, and gray matter hypometabolism in Alzheimer's disease.
In early Alzheimer's disease (AD), the hippocampal region is the area most severely affected by cellular and structural alterations, yet glucose hypometabolism predominates in the posterior association cortex and posterior cingulate gyrus. One prevalent hypothesis to account for this discrepancy is that posterior cingulate hypometabolism results from disconnection from the hippocampus through disruption of the cingulum bundle. However, only partial and indirect evidence currently supports this hypothesis. Thus, using structural magnetic resonance imaging and 2-[(18)F]fluoro-2-deoxy-D-glucose positron emission tomography in 18 patients with early AD, we assessed the relationships between hippocampal atrophy, white matter integrity, and gray matter metabolism by means of a whole-brain voxel-based correlative approach. We found that hippocampal atrophy is specifically related to cingulum bundle disruption, which is in turn highly correlated to hypometabolism of the posterior cingulate cortex but also of the middle cingulate gyrus, thalamus, mammillary bodies, parahippocampal gyrus, and hippocampus (all part of Papez's circuit), as well as the right temporoparietal associative cortex. These results provide the first direct evidence supporting the disconnection hypothesis as a major factor contributing to the early posterior hypometabolism in AD. Disruption of the cingulum bundle also appears to relate to hypometabolism in a large connected network over and above the posterior cingulate cortex, encompassing the whole memory circuit of Papez (consistent with the key location of this white matter tract within this loop) and also, but indirectly, the right posterior association cortex.


A study of some factors affecting the efficiency of in vitro mussel tissue culture for chromosome preparation.
In the course of a series of cultures of mussel mantle tissue three types of limiting factors to the efficiency of the method were identified. They are concerned with medium composition, and tissue culture and explant harvesting procedures. To reduce their negative effects, three changes with respect to the original technique are proposed: (1) incorporation of chick embryo extract in the place of egg yolk for culture medium enrichment; (2) replacement of 1 mm2 explants by 4 mm2 explants so as to suppress centrifugation, thereby gaining time; and (3) avoiding the use of male mantle tissue when it contains mature spermatozoa since this is cumbersome for slide analysis. Conversely, female mantle tissue can be employed without any disadvantage throughout the year especially as mature oocytes give suitable metaphase plates for meiotic chromosome study.


Simulation of the strong interaction.
In the Standard Model (SM) of particle physics, quarks are permanently confined by the strong interaction into bound states called hadrons. The values of some parameters, such as the quark masses and the strengths of the decays of one quark flavour into another, cannot be measured directly and must be deduced from experiments on hadrons. This requires calculations of the strong-interaction effects within the bound states, which are only possible using numerical simulations of quantum chromodynamics (QCD), the quantum field theory of the strong interaction. In conjunction with experimental data from B factories over the next few years, QCD simulations may provide clues to physics beyond the SM. The simulations are computationally intensive and, for the past 20 years, have exploited leading-edge computing technology. This continues today, with a project to develop a 10 Tflops computer for QCD costing less than 1 US dollar per Mflops.