Mangiferin induces islet regeneration in aged mice through regulating p16INK4a.
Previous studies by our group on mangiferin demonstrated that it exerts an anti‑hyperglycemic effect through the regulation of cell cycle proteins in 3‑month‑old, partially pancreatectomized (PPx) mice. However, β‑cell proliferation is known to become severely restricted with advanced age. Therefore, it is unknown whether mangiferin is able to reverse the diabetic condition and retain β‑cell regeneration capability in aged mice. In the present study, 12‑month‑old C57BL/6J mice that had undergone PPx were subjected to mangiferin treatment (90 mg/kg) for 28 days. Mangiferin‑treated aged mice exhibited decreased blood glucose levels and increased glucose tolerance, which was accompanied with higher serum insulin levels when compared with those in untreated PPx control mice. In addition, islet hyperplasia, elevated β‑cell proliferation and reduced β‑cell apoptosis were also identified in the mice that received mangiferin treatment. Further studies on the mRNA transcript and protein expression levels indicated comparatively increased levels of cyclins D1 and D2 and cyclin‑dependent kinase 4 in mangiferin‑treated mice, while the levels of p27Kip1 and p16INK4a were decreased relative to those in the untreated PPx controls. Of note, mangiferin treatment improved the proliferation rate of islet β‑cells in adult mice overexpressing p16INK4a, suggesting that mangiferin induced β‑cell proliferation via the regulation of p16INK4a. In addition, the mRNA transcription levels of critical genes associated with insulin secretion, including pancreatic and duodenal homeobox 1, glucose transporter 2 and glucokinase, were observed to be upregulated after mangiferin treatment. Taken together, it was indicated that mangiferin treatment significantly induced β‑cell proliferation and inhibited β‑cell apoptosis by regulating cell cycle checkpoint proteins. Furthermore, mangiferin was also demonstrated to regulate genes associated with insulin secretion. Collectively these, results suggest the therapeutic potential of mangiferin in the treatment of diabetes in aged individuals.


Developmental regulation of Bcl-2 family protein expression in the involuting mammary gland.
Epithelial cells within the mammary gland undergo developmental programmes of proliferation and apoptosis during the pregnancy cycle. After weaning, secretory epithelial cells are removed by apoptosis. To determine whether members of the Bcl-2 gene family could be involved in regulating this process, we have examined whether changes in their expression occur during this developmental apoptotic program in vivo. Bax and Bcl-x were evenly expressed throughout development. However, expression of Bak and Bad was increased during late pregnancy and lactation, and the proteins were present during the time of maximal apoptotic involution. Thereafter, their levels declined. In contrast, Bcl-w was expressed in pregnancy and lactation but was downregulated at the onset of apoptosis. Bcl-2 was not detected in lactating or early involuting mammary gland. Thus, the pro-apoptotic proteins Bax, Bak and Bad, as well as the death-suppressors Bcl-x, Bcl-2 and Bcl-w, are synthesised in mouse mammary gland, and dynamic changes in the expression profiles of these proteins occurs during development. To determine if changes in Bak and Bcl-w expression could regulate mammary apoptosis, their effect on cultured mouse mammary epithelial cells was examined in transient transfection assays. Enforced expression of Bak induced rapid mammary apoptosis, which could be suppressed by coexpression of Bcl-w. In extracts of mammary tissue in vivo, Bak heterodimerized with Bcl-x whereas Bax associated with Bcl-w, but Bak/Bcl-w heterodimers were not detected. Thus, Bak and Bcl-w may regulate cell death through independent pathways. These results support a model in which mammary epithelial cells are primed for apoptosis during the transition from pregnancy to lactation by de novo expression of the death effectors Bak and Bad. It is suggested that these proteins are prevented from triggering apoptosis by anti-apoptotic Bcl-2 family proteins until involution, when the levels of Bcl-w decline. Our study provides evidence that regulated changes in the expression of cell death genes may contribute to the developmental control of mammary apoptosis.


The impact of climate variability and change on the energy system: A systematic scoping review.
The energy system is a vital infrastructure which can be vulnerable to climate variability and change (CV&C) impacts. Understanding the impacts can prevent disruption and inform policy decision making. This study applied a scoping review in a systematic manner following the Joanna Briggs Institute guidelines to identify consistent patterns of CV&C impacts on the energy system, map and locate research gaps in the literature. A total of 176 studies were identified as eligible for inclusion in the review. This study found evidence of consistent increase in energy demand for Africa, the Americas and Asian continent. Consistent decrease was found in Northern and Eastern Europe, while increase in residential demand was projected in Oceania. There was evidence of consistent decrease in thermal power plant output globally. Solar photovoltaic showed a robust consistent pattern of increase in the Caribbean and Central America, Northern and Southern Africa and Oceania. As the global climate is changing in a future that is highly uncertain, the energy system should also evolve in order to adapt to the changing climate. Future impact assessment must integrate the impact of CV&C on power demand and supply while consider socioeconomic dynamics, cross-sectoral linkages and back-loops in a complete energy system model.


Sustainability of return to work in sick-listed employees with low-back pain. Two-year follow-up in a randomized clinical trial comparing multidisciplinary and brief intervention.
Sick-listed employees with low back pain had similar return to work (RTW) rates at one-year follow-up in a randomized trial comparing two interventions, but the effects were modified by specific workplace related factors. The present study addressed the sustainability of the intervention effects by performing a two-year follow-up and by using different outcome measures. A total of 351 employees sick-listed for 3-16 weeks due to LBP were recruited from their general practitioners and were randomly allocated to a hospital-based brief or multidisciplinary intervention. Outcome measures were based on sick leave registered in a national database of social and health-related benefits. RTW rates, RTW status, sick leave weeks and sick leave relapse were studied. During the two-year follow-up 80.0% and 77.3% had RTW for at least four weeks continuously, and the percentages with RTW at the 104th week were 61.1% and 58.0% in the brief and multidisciplinary intervention groups, respectively. At the 104th week 16.6% and 18.8% were on sick leave in the two groups, respectively, and about 12% were employed in modified jobs or participated in job training. The number of weeks on sick leave in the first year was significantly lower in the brief intervention group (median 14 weeks) than in the multidisciplinary intervention group (median 20 weeks), but during the second year the number of weeks on sick leave were not significantly different between intervention groups. Subgroups characterised by specific work related factors modified the effect of the intervention groups on RTW rates (p = 0.017). No difference in sick leave relapse was found between the intervention groups. The effects of the brief and multidisciplinary interventions at the two-year follow-up were in general similar to the effects at one-year follow-up. Current Controlled Trials ISRCTN18609003.


Exposure to Trauma and Mental Health Service Engagement Among Adults Who Were Children of the Northern Ireland Troubles of 1968 to 1998.
Northern Ireland is an appropriate region to examine the impact of traumatic experiences, owing to the many years of civil violence that have characterized its recent history, known colloquially as the "Troubles." Given the prominence of traumatic experiences among the aging population of Northern Ireland (NI), an evidence base is required to inform the planning and provision of effective mental health and other services. We analyzed the follow-up interviews (n = 225) of individuals from the Northern Ireland Study of Health and Stress (NISHS), aged 45 years and older, who experienced one or more conflict-related traumatic events. This study demonstrated that in NI traumatic events, such as being involved in an explosion, seeing someone killed or seriously injured, and living in a region of terror were most likely to be related to the Troubles. However, event types that we had not previously known to be related to conflict (such as the sudden death of a loved one), were also often related to the Troubles. Two-thirds of participants (67.1%) reported exposure to a Troubles-related event, and 57.8% reported being a civilian in a region of terror. The vast majority (85.9%) of participants who experienced a Troubles-related trauma never sought help, despite 59.1% meeting the criteria for a lifetime mental disorder. The reasons for not seeking help and sources of help are outlined. Policy makers must address Troubles-related mental health effects, in terms of how they carry forward into aging, and consider ways of improving engagement with services and treatments.


Effect of the Veterans Affairs Medical System on plastic surgery residency training.
Teaching hospitals within the Veterans Affairs Health System perform the majority of complex and high-risk surgical procedures in the veteran patient population. Residency positions in the Veterans Affairs Medical System are usually part of a rotational educational system within a university-based residency, and plastic surgeons in training are a major work force and health care provider. The purpose of this study was to evaluate the current effect of the Veterans Affairs Medical System on plastic surgery residency training. A 6-year (January of 1998 to December of 2003) review was performed of procedures completed at the Durham Veterans Affairs Medical Center, Section of Plastic Surgery. Procedures were divided into the following categories: extremities and trunk; breast and cosmetic; head and neck, including excision of skin lesions; hand surgery; craniomaxillofacial surgery; and other. Only procedures performed in the main operating room were reviewed and analyzed. In addition, a detailed review was performed of major head and neck reconstructions with free tissue transfer. A total of 1655 operative procedures were performed in 1290 patients. The ratio of men to women was 6:1 (1112 men and 178 women). Patients ranged in age from 26 to 97 years (average age, 62.7 years). Procedures in the extremities and trunk (n = 193, 11.7 percent), breast and cosmetic (n = 228, 13.8 percent), hand surgery (n = 155, 9.4 percent), and other (n = 275, 16.6 percent) categories were comparably distributed. Although the head and neck category accounted for the highest number of procedures (n = 766, 46.3 percent), the majority of these procedures were simple excisions of skin tumors (n = 612). There were significantly fewer major craniomaxillofacial cases (n = 38, 2.3 percent). Data from the retrospective analysis reveal that a broad spectrum of plastic surgical procedures is performed within the Veterans Affairs Health System, serving as a tremendous resource for resident training. The fact that approximately 260 procedures per year are performed demonstrates an active service. Craniomaxillofacial surgery is currently underrepresented compared with other categories at the authors' particular institution. By their definition, nonspecific plastic surgical procedures ("other") account for 275 (16.6 percent) of all 1655 procedures performed. This demonstrates that plastic surgery "overlaps" with other specialties, such as dermatology. The Veterans Affairs Health System will continue to play a significant role in the future training of plastic surgeons. Now, more than ever, a strong Veterans Affairs surgical service, including plastic surgery and its modern techniques, will be needed.


rBCG induces strong antigen-specific T cell responses in rhesus macaques in a prime-boost setting with an adenovirus 35 tuberculosis vaccine vector.
BCG vaccination, combined with adenoviral-delivered boosts, represents a reasonable strategy to augment, broaden and prolong immune protection against tuberculosis (TB). We tested BCG (SSI1331) (in 6 animals, delivered intradermally) and a recombinant (rBCG) AFRO-1 expressing perfringolysin (in 6 animals) followed by two boosts (delivered intramuscullary) with non-replicating adenovirus 35 (rAd35) expressing a fusion protein composed of Ag85A, Ag85B and TB10.4, for the capacity to induce antigen-specific cellular immune responses in rhesus macaques (Macaca mulatta). Control animals received diluent (3 animals). Cellular immune responses were analyzed longitudinally (12 blood draws for each animal) using intracellular cytokine staining (TNF-alpha, IL-2 and IFN-gamma), T cell proliferation was measured in CD4(+), CD8alpha/beta(+), and CD8alpha/alpha(+) T cell subsets and IFN-gamma production was tested in 7 day PBMC cultures (whole blood cell assay, WBA) using Ag85A, Ag85B, TB10.4 recombinant proteins, PPD or BCG as stimuli. Animals primed with AFRO-1 showed i) increased Ag85B-specific IFN-gamma production in the WBA assay (median >400 pg/ml for 6 animals) one week after the first boost with adenoviral-delivered TB-antigens as compared to animals primed with BCG (<200 pg/ml), ii) stronger T cell proliferation in the CD8alpha/alpha(+) T cell subset (proliferative index 17%) as compared to BCG-primed animals (proliferative index 5% in CD8alpha/alpha(+) T cells). Polyfunctional T cells, defined by IFN-gamma, TNF-alpha and IL-2 production were detected in 2/6 animals primed with AFRO-1 directed against Ag85A/b and TB10.4; 4/6 animals primed with BCG showed a Ag85A/b responses, yet only a single animal exhibited Ag85A/b and TB10.4 reactivity. AFRO-1 induces qualitatively and quantitatively different cellular immune responses as compared with BCG in rhesus macaques. Increased IFN-gamma-responses and antigen-specific T cell proliferation in the CD8alpha/alpha+ T cell subset represents a valuable marker for vaccine-take in BCG-based TB vaccine trials.


FoodOn: a harmonized food ontology to increase global food traceability, quality control and data integration.
The construction of high capacity data sharing networks to support increasing government and commercial data exchange has highlighted a key roadblock: the content of existing Internet-connected information remains siloed due to a multiplicity of local languages and data dictionaries. This lack of a digital lingua franca is obvious in the domain of human food as materials travel from their wild or farm origin, through processing and distribution chains, to consumers. Well defined, hierarchical vocabulary, connected with logical relationships-in other words, an ontology-is urgently needed to help tackle data harmonization problems that span the domains of food security, safety, quality, production, distribution, and consumer health and convenience. FoodOn (http://foodon.org) is a consortium-driven project to build a comprehensive and easily accessible global farm-to-fork ontology about food, that accurately and consistently describes foods commonly known in cultures from around the world. FoodOn addresses food product terminology gaps and supports food traceability. Focusing on human and domesticated animal food description, FoodOn contains animal and plant food sources, food categories and products, and other facets like preservation processes, contact surfaces, and packaging. Much of FoodOn's vocabulary comes from transforming LanguaL, a mature and popular food indexing thesaurus, into a World Wide Web Consortium (W3C) OWL Web Ontology Language-formatted vocabulary that provides system interoperability, quality control, and software-driven intelligence. FoodOn compliments other technologies facilitating food traceability, which is becoming critical in this age of increasing globalization of food networks.


De-novo versus recurrent hepatocellular carcinoma following direct-acting antiviral therapy for hepatitis C virus.
A recent appearance of direct-acting antivirals (DAAs) led to a surge in hepatitis C virus (HCV) management. Nowadays, a large proportion of treated patients have cirrhosis with a retained possibility to develop hepatocellular carcinoma (HCC) even after complete cure. We aimed to study tumoral differences between patients who developed HCC after DAAs as either a recurrence or de-novo HCC. We retrospectively analyzed 89 patients who presented to our HCC multidisciplinary clinic with HCC lesions following DAA therapy. A total of 45 patients had complete response to HCC according to the modified Response Evaluation Criteria in Solid Tumors before DAAs intake. Another 44 patients developed de-novo lesions after DAA treatment. Both groups were compared regarding their baseline characteristics, tumor criteria, response to DAAs as well response to HCC treatment. Both groups showed no significant difference regarding their baseline characteristics (age, sex, Child-Pugh score, and performance status) or response to DAAs (P=0.5). No significant difference was present between groups according to number, site, and size of lesions. However, time elapsed between the end of DAAs therapy and first diagnosis of HCC was significantly longer in de-novo group (15.22±16.39 months) versus recurrence group (6.76±5.1 months) (P=0.008). In addition, response to ablation was significantly better in de-novo lesions compared with recurrent HCC (P=0.03). Although de-novo HCC lesions significantly developed later than recurrent lesions in DAAs-treated patients, their response rates were significantly better. No differences were detected between both groups in their response to DAAs and their tumoral characteristics.


Improving spatiotemporal resolution of USPIO-enhanced dynamic imaging of rat kidneys.
This paper addresses the problem of enhancing spatiotemporal resolution of ultra-small superparamagnetic iron oxide (USPIO)-enhanced dynamic MRI of rat kidneys. To alleviate the limited resolution problem of conventional full-scan Fourier imaging methods, we use a generalized series-based imaging scheme to reduce coverage of kappa-space. Experimental results demonstrate that the generalized series imaging method with basis functions constructed using two references (pre- and post-contrast) can reduce the number of phase encodings measured during the dynamic contrast wash-in process by a factor of 4 with a negligible or minimal loss of image quality. The method is expected to make 3D studies possible using USPIO-enhanced dynamic imaging of rat kidneys, and prove valuable for early detection of renal rejection after kidney transplantation.


Sulphur mineralization kinetics of cattle manure and green waste compost in soils.
Sulphur mineralization of cattle manure (CM) and green waste compost (GWC) added to six agricultural soils with different chemical properties was monitored over 10 weeks in a laboratory incubation experiment. Although the amount of sulphur was higher in CM than in GWC, the cumulative SO4(2-)-S values in GWC-treated soils were higher than in soil amended with CM. The percentages of mineralized S were always higher in GWC-treated soil (in the range 1.3-8.5%) than in CM-treated soil (in the range 0.9-3.8%). In three of the six soils, particularly for CM, an immobilization of sulphur was observed. Three kinetic models were evaluated for their suitability to describe the mineralization process. The first-order model best described S mineralization for both amended and control soils. The GWC substantially increased the amount of potentially mineralizable S (S0) relative to the controls. In GWC-treated soils, the rates of S mineralization (k) were higher than rates in the controls. The k of CM-amended soils was often lower than the k of control soils. Parameters derived from the model were tested as indices for assessing the relationships between S mineralization and soil characteristics. The S0 was positively correlated to the amount of cumulative SO4(2-)-S and also to the content of organic C, N and S in soil.


[Analysis of the Cochrane Review: Pedicle Screw Fixation for Traumatic Fractures of the Thoracic and Lumbar Spine. Cochrane Database Syst Rev. 2013;05:CD009073].
Traumatic fractures of the thoracic and lumbar spine are common causes of spine surgery. Pedicle screw fixation is usually chosen, using monosegmentar, short or long segment instrumentations, with or without bone graft. This review aims to evaluate the effect of transpedicular fixation in traumatic fractures of the thoracic and lumbar spine. A systematic search on controlled, randomized or quasi-randomized trials comparing different methods of surgical treatment of this fractures was performed, followed by a process of article selection, data extraction and bias assessment by 3 independent authors. Eight articles were included in a total of 5 comparisons, between different transpedicular fixation techniques. No significant differences on function or quality of life, neurologic status or limitation of motion were found. Only instrumentation with fracture level screw incorporation showed significant decrease of pain when compared with instrumentation alone. Several techniques resulted in significant improvements of different radiological parameters. Significantly, surgeries with smaller duration were associated with lesser blood loss. Bone graft use caused a significant raise in post-operative complications, namely donor site pain. So, this paper showed that significative improvements in radiological parameters do not associate with correspondent clinical benefits, and only instrumentation with level screw incorporation is associated with a clear benefit on pain. Moreover, the need for bone graft is questioned, since it leads to no clinic-radiological improvement with a raise of complications. However, a small number of controlled studies is available on this topic.


The Health Care Job Engine: Where Do They Come From and What Do They Say About Our Future?
Health care has been cited as a job engine for the U.S. economy. This study used the Current Population Survey to examine the sector and occupation shifts that underlie this growth trend. Health care has had a cyclical relationship with retail trade, leisure and hospitality, education, and professional services. The entering workforce has been increasingly taking on low-skilled occupations. The exiting workforce has not been necessarily retiring or going back to school, but appeared to be leaving without a job, with potentially more child care duties, and with high rates of disability and poverty levels. This study also found that the number of workers staying in health care has been slowly declining over time. As the United States moves toward team-based care, more attention should be paid to the needs of the lower skilled workers to reduce turnover and ensure delivery of quality care.


Cloning Should Be Simple: Escherichia coli DH5α-Mediated Assembly of Multiple DNA Fragments with Short End Homologies.
Numerous DNA assembly technologies exist for generating plasmids for biological studies. Many procedures require complex in vitro or in vivo assembly reactions followed by plasmid propagation in recombination-impaired Escherichia coli strains such as DH5α, which are optimal for stable amplification of the DNA materials. Here we show that despite its utility as a cloning strain, DH5α retains sufficient recombinase activity to assemble up to six double-stranded DNA fragments ranging in size from 150 bp to at least 7 kb into plasmids in vivo. This process also requires surprisingly small amounts of DNA, potentially obviating the need for upstream assembly processes associated with most common applications of DNA assembly. We demonstrate the application of this process in cloning of various DNA fragments including synthetic genes, preparation of knockout constructs, and incorporation of guide RNA sequences in constructs for clustered regularly interspaced short palindromic repeats (CRISPR) genome editing. This consolidated process for assembly and amplification in a widely available strain of E. coli may enable productivity gain across disciplines involving recombinant DNA work.


Cobalamin Deficiency in the Elderly.
Older people are at risk for cobalamin (vitamin B12) deficiency because of a number of common disorders (e.g., autoimmune gastritis) and drugs (e.g., antacids) that may alter its absorption and utilization. The prevalence of cobalamin deficiency increases with age, resulting, particularly elevated, in frail and institutionalized subjects. At variance with common sense, the diagnosis is far from simple. It requires a high degree of suspicion, due to heterogeneity and non-specificity of the signs and symptoms, ranging from macrocytosis (with or without anemia) to neuropsychiatric manifestations, that characterize several other aging-related disorders, like hematological malignancies, diabetes, hypothyroidism or vasculopathy. Furthermore, the detection of low levels of serum vitamin B12 appears poorly sensitive and specific. Other biomarkers, like serum homocysteine or methylmalonic acid, have improved the diagnostic possibilities but are expensive, not widely available, and may be influenced by some confounders (e.g., folate deficiency, or chronic renal failure). Early recognition and treatment are crucial since a proportion of patients develop severe complications, such as bone marrow failure and irreversible neurological impairment. High-dose oral treatment has proven to be as effective as the parenteral route, even in subjects with malabsorption, ensuring the complete resolution in the majority of cases. In this review, we trace the essential role of cobalamin in humans, the possible causes and impact of deficiency, the diagnostic challenges and the therapeutic options, between old and emerging concepts, with a particular focus on the elderly.


The effect of polyploidy and hybridization on the evolution of floral colour in Nicotiana (Solanaceae).
Speciation in angiosperms can be accompanied by changes in floral colour that may influence pollinator preference and reproductive isolation. This study investigates whether changes in floral colour can accompany polyploid and homoploid hybridization, important processes in angiosperm evolution. Spectral reflectance of corolla tissue was examined for 60 Nicotiana (Solanaceae) accessions (41 taxa) based on spectral shape (corresponding to pigmentation) as well as bee and hummingbird colour perception in order to assess patterns of floral colour evolution. Polyploid and homoploid hybrid spectra were compared with those of their progenitors to evaluate whether hybridization has resulted in floral colour shifts. Floral colour categories in Nicotiana seem to have arisen multiple times independently during the evolution of the genus. Most younger polyploids displayed an unexpected floral colour, considering those of their progenitors, in the colour perception of at least one pollinator type, whereas older polyploids tended to resemble one or both of their progenitors. Floral colour evolution in Nicotiana is weakly constrained by phylogeny, and colour shifts do occur in association with both polyploid and homoploid hybrid divergence. Transgressive floral colour in N. tabacum has arisen by inheritance of anthocyanin pigmentation from its paternal progenitor while having a plastid phenotype like its maternal progenitor. Potentially, floral colour evolution has been driven by, or resulted in, pollinator shifts. However, those polyploids that are not sympatric (on a regional scale) with their progenitor lineages are typically not divergent in floral colour from them, perhaps because of a lack of competition for pollinators.


Deformed wing virus can be transmitted during natural mating in honey bees and infect the queens.
Deformed wing virus is an important contributor to honey bee colony losses. Frequently queen failure is reported as a cause for colony loss. Here we examine whether sexual transmission during multiple matings of queens is a possible way of virus infection in queens. In an environment with high prevalence of deformed wing virus, queens (n = 30) were trapped upon their return from natural mating flights. The last drone's endophallus (n = 29), if present, was removed from the mated queens for deformed wing virus quantification, leading to the detection of high-level infection in 3 endophalli. After oviposition, viral quantification revealed that seven of the 30 queens had high-level deformed wing virus infections, in all tissues, including the semen stored in the spermathecae. Two groups of either unmated queens (n = 8) with induced egg laying, or queens (n = 12) mated in isolation with drones showing comparatively low deformed wing virus infections served as control. None of the control queens exhibited high-level viral infections. Our results demonstrate that deformed wing virus infected drones are competitive to mate and able to transmit the virus along with semen, which occasionally leads to queen infections. Virus transmission to queens during mating may be common and can contribute noticeably to queen failure.


Transcriptional regulation of vascular endothelial cell responses to hypoxia by HIF-1.
Hypoxia-inducible factor 1 (HIF-1) activates transcription of genes encoding angiogenic growth factors, which are secreted by hypoxic cells and stimulate endothelial cells, leading to angiogenesis. To determine whether HIF-1 also mediates cell-autonomous responses to hypoxia, we have compared gene expression profiles in arterial endothelial cells cultured under nonhypoxic versus hypoxic conditions and in nonhypoxic cells infected with adenovirus encoding beta-galactosidase versus a constitutively active form of HIF-1alpha (AdCA5). There were 245 gene probes that showed at least 1.5-fold increase in expression in response to hypoxia and in response to AdCA5; 325 gene probes showed at least 1.5-fold decrease in expression in response to hypoxia and in response to AdCA5. The largest category of genes down-regulated by both hypoxia and AdCA5 encoded proteins involved in cell growth/proliferation. Many genes up-regulated by both hypoxia and AdCA5 encoded cytokines/growth factors, receptors, and other signaling proteins. Transcription factors accounted for the largest group of HIF-1-regulated genes, indicating that HIF-1 controls a network of transcriptional responses to hypoxia in endothelial cells. Infection of endothelial cells with AdCA5 under nonhypoxic conditions was sufficient to induce increased basement membrane invasion and tube formation similar to the responses induced by hypoxia, indicating that HIF-1 mediates cell-autonomous activation of endothelial cells.


Comparison of amitriptyline supplemented with pregabalin, pregabalin supplemented with amitriptyline, and duloxetine supplemented with pregabalin for the treatment of diabetic peripheral neuropathic pain (OPTION-DM): a multicentre, double-blind, randomised crossover trial.
Diabetic peripheral neuropathic pain (DPNP) is common and often distressing. Most guidelines recommend amitriptyline, duloxetine, pregabalin, or gabapentin as initial analgesic treatment for DPNP, but there is little comparative evidence on which one is best or whether they should be combined. We aimed to assess the efficacy and tolerability of different combinations of first-line drugs for treatment of DPNP. OPTION-DM was a multicentre, randomised, double-blind, crossover trial in patients with DPNP with mean daily pain numerical rating scale (NRS) of 4 or higher (scale is 0-10) from 13 UK centres. Participants were randomly assigned (1:1:1:1:1:1), with a predetermined randomisation schedule stratified by site using permuted blocks of size six or 12, to receive one of six ordered sequences of the three treatment pathways: amitriptyline supplemented with pregabalin (A-P), pregabalin supplemented with amitriptyline (P-A), and duloxetine supplemented with pregabalin (D-P), each pathway lasting 16 weeks. Monotherapy was given for 6 weeks and was supplemented with the combination medication if there was suboptimal pain relief (NRS >3), reflecting current clinical practice. Both treatments were titrated towards maximum tolerated dose (75 mg per day for amitriptyline, 120 mg per day for duloxetine, and 600 mg per day for pregabalin). The primary outcome was the difference in 7-day average daily pain during the final week of each pathway. This trial is registered with ISRCTN, ISRCTN17545443. Between Nov 14, 2017, and July 29, 2019, 252 patients were screened, 140 patients were randomly assigned, and 130 started a treatment pathway (with 84 completing at least two pathways) and were analysed for the primary outcome. The 7-day average NRS scores at week 16 decreased from a mean 6·6 (SD 1·5) at baseline to 3·3 (1·8) at week 16 in all three pathways. The mean difference was -0·1 (98·3% CI -0·5 to 0·3) for D-P versus A-P, -0·1 (-0·5 to 0·3) for P-A versus A-P, and 0·0 (-0·4 to 0·4) for P-A versus D-P, and thus not significant. Mean NRS reduction in patients on combination therapy was greater than in those who remained on monotherapy (1·0 [SD 1·3] vs 0·2 [1·5]). Adverse events were predictable for the monotherapies: we observed a significant increase in dizziness in the P-A pathway, nausea in the D-P pathway, and dry mouth in the A-P pathway. To our knowledge, this was the largest and longest ever, head-to-head, crossover neuropathic pain trial. We showed that all three treatment pathways and monotherapies had similar analgesic efficacy. Combination treatment was well tolerated and led to improved pain relief in patients with suboptimal pain control with a monotherapy. National Institute for Health Research (NIHR) Health Technology Assessment programme.


Development and pilot testing of a mental healthcare plan in Nepal.
Mental health service delivery models that are grounded in the local context are needed to address the substantial treatment gap in low- and middle-income countries. To present the development, and content, of a mental healthcare plan (MHCP) in Nepal and assess initial feasibility. A mixed methods formative study was conducted. Routine monitoring and evaluation data, including client flow and reports of satisfaction, were obtained from patients (n = 135) during the pilot-testing phase in two health facilities. The resulting MHCP consists of 12 packages, divided over community, health facility and organisation platforms. Service implementation data support the real-life applicability of the MHCP, with reasonable treatment uptake. Key barriers were identified and addressed, namely dissatisfaction with privacy, perceived burden among health workers and high drop-out rates. The MHCP follows a collaborative care model encompassing community and primary healthcare interventions.


Evoked Potentials Reveal Noise Exposure-Related Central Auditory Changes Despite Normal Audiograms.
Purpose Complaints of auditory perceptual deficits, such as tinnitus and difficulty understanding speech in background noise, among individuals with clinically normal audiograms present a perplexing problem for audiologists. One potential explanation for these "hidden" auditory deficits is loss of the synaptic connections between the inner hair cells and their afferent auditory nerve fiber targets, a condition that has been termed cochlear synaptopathy. In animal models, cochlear synaptopathy can occur due to aging or exposure to noise or ototoxic drugs and is associated with reduced auditory brainstem response (ABR) wave I amplitudes. Decreased ABR wave I amplitudes have been demonstrated among young military Veterans and non-Veterans with a history of firearm use, suggesting that humans may also experience noise-induced synaptopathy. However, the downstream consequences of synaptopathy are unclear. Method To investigate how noise-induced reductions in wave I amplitude impact the central auditory system, the ABR, the middle latency response (MLR), and the late latency response (LLR) were measured in 65 young Veterans and non-Veterans with normal audiograms. Results In response to a click stimulus, the MLR was weaker for Veterans compared to non-Veterans, but the LLR was not reduced. In addition, low ABR wave I amplitudes were associated with a reduced MLR, but with an increased LLR. Notably, Veterans reporting tinnitus showed the largest mean LLRs. Conclusions These findings indicate that decreased peripheral auditory input leads to compensatory gain in the central auditory system, even among individuals with normal audiograms, and may impact auditory perception. This pattern of reduced MLR, but not LLR, was observed among Veterans even after statistical adjustment for sex and distortion product otoacoustic emission differences, suggesting that synaptic loss plays a role in the observed central gain. Supplemental Material https://doi.org/10.23641/asha.11977854.


Adult spinal cord progenitor cells are repelled by netrin-1 in the embryonic and injured adult spinal cord.
Adult neural progenitor cells (aNPCs) exhibit limited migration in vivo with the exception of the rostral migratory stream and injury-induced movement. Surprisingly little is known regarding those signals regulating attraction or inhibition of the aNPC. These studies demonstrate that aNPCs respond principally to a repulsive cue expressed at the embryonic floor plate (FP) and also the injured adult CNS. Adult spinal cord progenitor cells (aSCPs) were seeded onto organotypic slice preparations of the intact embryonic or injured adult spinal cord. Cell migration assays combined with genetic and molecular perturbation of FP-derived migration cues or aSCP receptors establish netrin-1 (Ntn-1) but not Slit-2, Shh, or Ephrin-B3 as the primary FP-derived repellant. When slices were prepared from injured spinal cord, aSCP migration away from the injury core was Ntn-1-dependent. These studies establish Ntn-1 as a critical regulator of aSCP migration in the intact and injured CNS.


Resealable, optically accessible, PDMS-free fluidic platform for ex vivo interrogation of pancreatic islets.
We report the design and fabrication of a robust fluidic platform built out of inert plastic materials and micromachined features that promote optimized convective fluid transport. The platform is tested for perfusion interrogation of rodent and human pancreatic islets, dynamic secretion of hormones, concomitant live-cell imaging, and optogenetic stimulation of genetically engineered islets. A coupled quantitative fluid dynamics computational model of glucose stimulated insulin secretion and fluid dynamics was first utilized to design device geometries that are optimal for complete perfusion of three-dimensional islets, effective collection of secreted insulin, and minimization of system volumes and associated delays. Fluidic devices were then fabricated through rapid prototyping techniques, such as micromilling and laser engraving, as two interlocking parts from materials that are non-absorbent and inert. Finally, the assembly was tested for performance using both rodent and human islets with multiple assays conducted in parallel, such as dynamic perfusion, staining and optogenetics on standard microscopes, as well as for integration with commercial perfusion machines. The optimized design of convective fluid flows, use of bio-inert and non-absorbent materials, reversible assembly, manual access for loading and unloading of islets, and straightforward integration with commercial imaging and fluid handling systems proved to be critical for perfusion assay, and particularly suited for time-resolved optogenetics studies.


Moderate increases in daily step count are associated with reduced IL6 and CRP in women with PCOS.
Aims Physical activity has been proposed to be an effective non-pharmacological method of reducing systemic inflammation and therefore may prove particularly efficacious for women with polycystic ovary syndrome (PCOS) who have been shown to have high levels of inflammation and an increased risk of type 2 diabetes (T2DM) and cardiovascular disease (CVD). Therefore, the aim of the present study was to assess whether modest changes in daily step count could significantly reduce levels of inflammatory markers in women with PCOS. Subjects and Methods Sixty-five women with PCOS were assessed at baseline and again at 6 months. All had been provided with an accelerometer and encouraged to increase activity levels. Multivariate linear regression analyses (adjusted for age, ethnicity, baseline step count, change in BMI and change in accelerometer wear-time) were used to assess changes in daily step count against clinical and research biomarkers of inflammation, CVD and T2DM. Results Mean step count/day at baseline was 6337 (±270). An increase in step count (by 1000 steps) was associated with a 13% reduction in IL6 (β: -0.81 ng/L; 95% CI, -1.37, -0.25, P = 0.005) and a 13% reduction in CRP (β: -0.68 mg/L; 95% CI, -1.30, -0.06, P = 0.033). Additionally, there was a modest decrease in BMI (β: 0.20 kg/m2; 95% CI, -0.38, -0.01, P = 0.038). Clinical markers of T2DM and CVD were not affected by increased step count. Conclusions Modest increases in step count/day can reduce levels of inflammatory markers in women with PCOS, which may reduce the future risk of T2DM and CVD.


Update on giant cell arteritis.
Giant cell arteritis (GCA) is a challenging condition to manage because of the potential for acute irreversible vision loss and corticosteroid-related morbidity. Recent developments offer the potential to improve both the assessment and treatment of patients. Vascular imaging is increasingly being used in the diagnostic algorithm for GCA. Results from recent vascular ultrasound and high-resolution cranial MRI studies have led some groups to suggest forgoing temporal artery biopsy (TAB) in selected patients. The treatment armamentarium has been enhanced with the addition of Tocilizumab, a monoclonal antibody that inhibits IL-6 and has been shown to be effective in sustaining glucocorticoid-free remission out to 52 weeks. New publications have provided guidance in how clinicians can interpret minimally inflamed biopsies and navigate the controversy about what role, if any, varicella zoster virus may play in the pathophysiology of GCA. Basic science developments have improved our understanding of the immunopathology of GCA including the role of Th1 and Th17 lymphocytes and mechanisms of arterial wall lymphocyte invasion. There have been significant recent advances in GCA, particularly in relation to imaging and treatment options. Longer term outcome data will help clarify how best to utilize them in routine clinical practice.


Grafting with Cryopreserved Amniotic Membrane versus Conservative Wound Care in Treatment of Pressure Ulcers: A Randomized Clinical Trial.
To compare the healing process of pressure ulcers treated with cryopreserved human amniotic membrane allograft and routine pressure ulcer care in our hospital. From January 2012 to December 2013, in a prospective randomized clinical trial (IRCT201612041335N2), 24 patients with second and third stage of pressure ulcers were enrolled in this study. All patients needed split-thickness skin grafts for pressure ulcer-wound coverage. Selected patients had symmetric ulcers on both upper and lower extremities. The patients were randomly divided into two groups: amnion and control. In the amnion group, the ulcer was covered with cryopreserved amniotic membrane and in the control group it was treated with local Dilantin powder application. The duration and success rate of complete healing was compared between the two groups. The study group was composed of 24 pressure ulcers in 24 patients (19 males and 5 females) with a mean age of 44±12.70 years. The demographic characteristics, ulcer area, and underlying diseases were similar in both groups. The early sign of response, such as decrease in wound discharge, was detected 12-14 days after biological dressing. Complete pressure ulcer healing occurred only in the amnion group (p< 0.001). Partial healing was significantly higher in the amnion group (p< 0.03). Healing time in this group was faster than that the control group (20 days versus 54 days). No major complication was recorded with amniotic membrane dressing. Cryopreserved amniotic membrane is an effective biologic dressing that promotes re-epithelialization in pressure ulcers.


High Dynamic Range Pixel Array Detector for Scanning Transmission Electron Microscopy.
We describe a hybrid pixel array detector (electron microscope pixel array detector, or EMPAD) adapted for use in electron microscope applications, especially as a universal detector for scanning transmission electron microscopy. The 128×128 pixel detector consists of a 500 µm thick silicon diode array bump-bonded pixel-by-pixel to an application-specific integrated circuit. The in-pixel circuitry provides a 1,000,000:1 dynamic range within a single frame, allowing the direct electron beam to be imaged while still maintaining single electron sensitivity. A 1.1 kHz framing rate enables rapid data collection and minimizes sample drift distortions while scanning. By capturing the entire unsaturated diffraction pattern in scanning mode, one can simultaneously capture bright field, dark field, and phase contrast information, as well as being able to analyze the full scattering distribution, allowing true center of mass imaging. The scattering is recorded on an absolute scale, so that information such as local sample thickness can be directly determined. This paper describes the detector architecture, data acquisition system, and preliminary results from experiments with 80-200 keV electron beams.


Virtual drug repurposing study against SARS-CoV-2 TMPRSS2 target.
Currently, the world suffers from a new coronavirus SARS-CoV-2 that causes COVID-19. Therefore, there is a need for the urgent development of novel drugs and vaccines for COVID-19. Since it can take years to develop new drugs against this disease, here we used a hybrid combined molecular modeling approach in virtual drug screening repurposing study to identify new compounds against this disease. One of the important SARS-CoV-2 targets namely type 2 transmembrane serine protease (TMPRSS2) was screened with NPC's NIH small molecule library which includes approved drugs by FDA and compounds in clinical investigation. We used 6654 small molecules in molecular docking and top-50 docking scored compounds were initially used in short (10-ns) molecular dynamics (MD) simulations. Based on average MM/GBSA binding free energy results, long (100-ns) MD simulations were employed for the identified hits. Both binding energy results as well as crucial residues in ligand binding were also compared with a positive control TMPRSS2 inhibitor, Camostat mesylate. Based on these numerical calculations we proposed a compound (benzquercin) as strong TMPRSS2 inhibitor. If these results can be validated by in vitro and in vivo studies, benzquercin can be considered to be used as inhibitor of TMPRSS2 at the clinical studies.


Mass spectrometry-based identification of native cardiac Nav1.5 channel α subunit phosphorylation sites.
Cardiac voltage-gated Na+ (Nav) channels are key determinants of action potential waveforms, refractoriness and propagation, and Nav1.5 is the main Nav pore-forming (α) subunit in the mammalian heart. Although direct phosphorylation of the Nav1.5 protein has been suggested to modulate various aspects of Nav channel physiology and pathophysiology, native Nav1.5 phosphorylation sites have not been identified. In the experiments here, a mass spectrometry (MS)-based proteomic approach was developed to identify native Nav1.5 phosphorylation sites directly. Using an anti-NavPAN antibody, Nav channel complexes were immunoprecipitated from adult mouse cardiac ventricles. The MS analyses revealed that this antibody immunoprecipitates several Nav α subunits in addition to Nav1.5, as well as several previously identified Nav channel associated/regulatory proteins. Label-free comparative and data-driven phosphoproteomic analyses of purified cardiac Nav1.5 protein identified 11 phosphorylation sites, 8 of which are novel. All the phosphorylation sites identified except one in the N-terminus are in the first intracellular linker loop, suggesting critical roles for this region in phosphorylation-dependent cardiac Nav channel regulation. Interestingly, commonly used prediction algorithms did not reliably predict these newly identified in situ phosphorylation sites. Taken together, the results presented provide the first in situ map of basal phosphorylation sites on the mouse cardiac Nav1.5 α subunit.


Habitat selection responses of parents to offspring predation risk: an experimental test.
The ability of nest predation to influence habitat settlement decisions in birds is widely debated, despite its importance in limiting fitness. Here, we experimentally manipulated nest predation risk across a landscape and asked the question, do migratory birds assess and respond to variation in nest predation risk when choosing breeding habitats? We examined habitat preference by quantifying the density and settlement date of eight species of migratory passerines breeding in areas with and without intact nest predator communities. We found consistently more individuals nesting in areas with reduced nest predation than in areas with intact predator assemblages, although predation risk had no influence on settlement or breeding phenology. Additionally, those individuals occupying safer nesting habitats exhibited increased singing activity. These findings support a causal relationship between habitat choice and nest predation risk and suggest the importance of nest predation risk in shaping avian community structure and breeding activity.


Timely versus delayed early childhood vaccination and seizures.
Little is known regarding the timing of childhood vaccination and postvaccination seizures. In a cohort of 323 247 US children from the Vaccine Safety Datalink born from 2004 to 2008, we analyzed the association between the timing of childhood vaccination and the first occurrence of seizure with a self-controlled case series analysis of the first doses of individual vaccines received in the first 2 years of life. In infants, there was no association between the timing of infant vaccination and postvaccination seizures. In the second year of life, the incident rate ratio (IRR) for seizures after receipt of the first measles-mumps-rubella vaccine (MMR) dose at 12 to 15 months was 2.65 (95% confidence interval [CI] 1.99-3.55); the IRR after an MMR dose at 16 to 23 months was 6.53 (95% CI 3.15-13.53). The IRR for seizures after receipt of the first measles-mumps-rubella-varicella vaccine (MMRV) dose at 12 to 15 months was 4.95 (95% CI 3.68-6.66); the IRR after an MMRV dose at 16 to 23 months was 9.80 (95% CI 4.35 -22.06). There is no increased risk of postvaccination seizure in infants regardless of timing of vaccination. In year 2, delaying MMR vaccine past 15 months of age results in a higher risk of seizures. The strength of the association is doubled with MMRV vaccine. These findings suggest that on-time vaccination is as safe with regard to seizures as delayed vaccination in the first year of life, and that delayed vaccination in the second year of life is associated with more postvaccination seizures than on-time vaccination.


Effect of accounting for different phenotypic variances by sire breed and sex on selection of sires based on expected progeny differences for 200- and 365-day weights.
The effects of accounting for different phenotypic variances according to sire breed and sex subclasses on estimation of sire breed effects and prediction of expected progeny differences of sires mated to Hereford and Angus cows were investigated. Data consisted of 6,977 and 6,530 records of 200-d (weaning) and 365-d (yearling) weights, respectively, of F1 calves sired by bulls (662 and 661, respectively) of 23 breeds that have been evaluated in the Germ Plasm Evaluation Program at the U.S. Meat Animal Research Center, Clay Center, NE. Models compared included fixed effects of genetic group of sire (samples of sires evaluated at different times), dam breed, sex, birth year of calf and age of dam, plus sire within genetic group and dam within dam breed as random effects. Variance structures were different: Model I assumed homogeneous variances across sire breed-sex subclasses; Model II accounted for differences in phenotypic variance by sire breed and sex subclasses. Differences between estimates of sire group effects obtained with the two models were not significant for either trait. Product-moment and rank correlations between expected progeny differences obtained with Model I and Model II were greater than .93 when computed within each group and .99 or larger when computed across breeds. There were slight changes in the numbers of sires contributed by different breeds to the proportions selected across breeds under different selection intensities when sires were ranked with the two models. However, differences between means predicted under Model II were small when sires were ranked and selected based on the two models. Changes in standard errors of prediction for expected progeny differences and standard errors for estimates of breed effects obtained when adjusting for differences of phenotypic variances, compared to not adjusting, were proportional to the ratios of the phenotypic standard deviations of the sire breeds to the common phenotypic standard deviation.


Carcinoid tumor of the common bile duct: a rare complication of von Hippel-Lindau syndrome.
Von Hippel-Lindau syndrome (VHL) is a rare autosomal-dominant, inherited familial cancer syndrome. Hemangioblastomas, pheochromocytomas and renal carcinoma are the frequent reported VHL tumors. Neuroendocrine tumors have also been described, mostly in the pancreas and rarely in the biliary trees. We report the second case of bile duct carcinoid in a 31-year-old VHL woman. She underwent right adrenalectomy for a pheochromocytoma in the past. She also had a positive family history of phenotypic expression of VHL syndrome. The patient presented with biliary colic. Endoscopic retrograde cholangio-pancreatography showed intra-luminal bile duct mass. Surgical exploration identified a beige nodular lesion that was a carcinoid tumor on histology. This new association should be clarified by further genetic investigations.


Guidelines for the Management of Pediatric Severe Traumatic Brain Injury, Third Edition: Update of the Brain Trauma Foundation Guidelines, Executive Summary.
The purpose of this work is to identify and synthesize research produced since the second edition of these Guidelines was published and incorporate new results into revised evidence-based recommendations for the treatment of severe traumatic brain injury in pediatric patients. This document provides an overview of our process, lists the new research added, and includes the revised recommendations. Recommendations are only provided when there is supporting evidence. This update includes 22 recommendations, nine are new or revised from previous editions. New recommendations on neuroimaging, hyperosmolar therapy, analgesics and sedatives, seizure prophylaxis, temperature control/hypothermia, and nutrition are provided. None are level I, three are level II, and 19 are level III. The Clinical Investigators responsible for these Guidelines also created a companion algorithm that supplements the recommendations with expert consensus where evidence is not available and organizes possible interventions into first and second tier utilization. The purpose of publishing the algorithm as a separate document is to provide guidance for clinicians while maintaining a clear distinction between what is evidence based and what is consensus based. This approach allows, and is intended to encourage, continued creativity in treatment and research where evidence is lacking. Additionally, it allows for the use of the evidence-based recommendations as the foundation for other pathways, protocols, or algorithms specific to different organizations or environments. The complete guideline document and supplemental appendices are available electronically from this journal. These documents contain summaries and evaluations of all the studies considered, including those from prior editions, and more detailed information on our methodology. New level II and level III evidence-based recommendations and an algorithm provide additional guidance for the development of local protocols to treat pediatric patients with severe traumatic brain injury. Our intention is to identify and institute a sustainable process to update these Guidelines as new evidence becomes available.


Interaction between warfarin and cranberry juice.
Warfarin is extensively used for anticoagulation to a target international normalized ratio of 2.0-3.0 for most indications or 2.5-3.5 for high-risk indications; however, many drugs and dietary supplements induce fluctuations in the international normalized ratio. Such fluctuations may lead to therapeutic failure or bleeding complications. Cranberry juice is increasingly used for the prevention and adjunctive treatment of urinary tract infections. The United Kingdom's Committee on Safety of Medicines has alerted clinicians to a potential interaction between warfarin and cranberry juice and has advised that patients avoid their concurrent use. Review and analysis of the literature revealed that ingestion of large volumes of cranberry juice destabilize warfarin therapy. Small amounts of juice are not expected to cause such an interaction. Clinicians should be aware of this potential interaction and monitor and counsel patients accordingly.


Utilization of ICDS program by adolescent girls and implementation barriers in Urban Rishikesh, India.
Adolescent girls are at higher risk of mortality and morbidity due to childbirth. Government of India therefore initiated Adolescent girls' scheme since year 2000. However, since its inception program received various setbacks. This study was carried out to assess the utilization of ICDS program for adolescent girls through Anganwadi centers (AWC) and implementation barrier from providers point of view. Also study assessed gaps in knowledge, attitude, and practice of anemia. Mixed-method sequential explanatory approach was adopted. Quantitative data was collected by community-based cross-sectional survey in Urban Rishikesh, Uttarakhand. This was followed by in-depth interview of Anganwadi worker (AWW). Twenty adolescent girls from each of 20 clusters were selected by cluster sampling. Data was collected by structured validated questionnaire using Epicollect 5. AWWs were interviewed with help of interview guide. Variables were described as proportion and mean. The interview transcript was analyzed using content analysis. Out of 400 adolescent girls, 10% were school dropouts and 59% were undernourished. Seven percent had ever visited AWC in last year. The main reason for nonutilization was unawareness. 38% had knowledge regarding symptoms of anemia. AWW faces problems due to nonavailability of iron tablet, untrained for health education sessions, and service not meeting demand of beneficiaries. Poor utilization of ICDS services by adolescent girls due to unawareness. There is gap in knowledge regarding anemia among adolescent girls. Adolescent girls' scheme is not as per felt need of beneficiary and poorly implemented.


Time invariance of three ultra-brief internet-related instruments: Smartphone Application-Based Addiction Scale (SABAS), Bergen Social Media Addiction Scale (BSMAS), and the nine-item Internet Gaming Disorder Scale- Short Form (IGDS-SF9) (Study Part B).
Given the many technological advances over the past two decades, a small minority of young people are at risk of problematic use or becoming addicted to these technologies (including activities on the internet and smartphones). Many brief psychometric scales have been developed to assess those at risk of problematic use or addiction including the six-item Smartphone Application-Based Addiction Scale [SABAS], the six-item Bergen Social Media Addiction Scale [BSMAS], and the nine-item Internet Gaming Disorder Scale-Short Form [IGDS-SF9]). However, to date, the reproducibility of these three scales has only been examined over a short period of time (e.g., two weeks), and it is unclear whether they are time invariant across a longer period (e.g., three months). Given the emergence of internet and smartphone addiction in Chinese population, the present study translated the three instruments into Chinese and recruited 640 university students (304 from Hong Kong [99 males] and 336 from Taiwan [167 males]) to complete the three scales twice (baseline and three months after baseline). Multigroup confirmatory factor analysis (MGCFA) was applied to examine the time invariance. The intraclass correlation coefficient (ICC) was used to assess the relative reliability, and the percentage of smallest real difference (SRD%) was utilized to explore the absolute reliability for the three scales. MGCFA showed that all three scales were time invariant across three months. ICC demonstrated that all the scales were satisfactory in reproducibility (0.82 to 0.94), and SRD% indicated that all the scales had acceptable measurement noise (23.8 to 29.4). In conclusion, the short, valid, reliable, and easy-to-use Chinese SABAS, BSMAS, and IGDS-SF9 show good properties across periods of three months.


Provision of foot health services for people with rheumatoid arthritis in New South Wales: a web-based survey of local podiatrists.
It is unclear if podiatric foot care for people with rheumatoid arthritis (RA) in New South Wales (NSW) meets current clinical recommendations. The objective of this study was to survey podiatrists' perceptions of the nature of podiatric foot care provision for people who have RA in NSW. An anonymous, cross-sectional survey with a web-based questionnaire was conducted. The survey questionnaire was developed according to clinical experience and current foot care recommendations. State registered podiatrists practising in the state of NSW were invited to participate. The survey link was distributed initially via email to members of the Australian Podiatry Association (NSW), and distributed further through snowballing techniques using professional networks. Data was analysed to assess significant associations between adherence to clinical practice guidelines, and private/public podiatry practices. 86 podiatrists participated in the survey (78% from private practice, 22% from public practice). Respondents largely did not adhere to formal guidelines to manage their patients (88%). Only one respondent offered a dedicated service for patients with RA. Respondents indicated that the primary mode of accessing podiatry was by self-referral (68%). Significant variation was observed regarding access to disease and foot specific assessments and treatment strategies. Assessment methods such as administration of patient reported outcome measures, vascular and neurological assessments were not conducted by all respondents. Similarly, routine foot care strategies such as prescription of foot orthoses, foot health advice and footwear were not employed by all respondents. The results identified issues in foot care provision which should be explored through further research. Foot care provision in NSW does not appear to meet the current recommended standards for the management of foot problems in people who have RA. Improvements to foot care could be undertaken in terms of providing better access to examination techniques and treatment strategies that are recommended by evidence based treatment paradigms.


Intellectual contributions meriting authorship: Survey results from the top cited authors across all science categories.
Authorship is the currency of an academic career for which the number of papers researchers publish demonstrates creativity, productivity, and impact. To discourage coercive authorship practices and inflated publication records, journals require authors to affirm and detail their intellectual contributions but this strategy has been unsuccessful as authorship lists continue to grow. Here, we surveyed close to 6000 of the top cited authors in all science categories with a list of 25 research activities that we adapted from the National Institutes of Health (NIH) authorship guidelines. Responses varied widely from individuals in the same discipline, same level of experience, and same geographic region. Most researchers agreed with the NIH criteria and grant authorship to individuals who draft the manuscript, analyze and interpret data, and propose ideas. However, thousands of the researchers also value supervision and contributing comments to the manuscript, whereas the NIH recommends discounting these activities when attributing authorship. People value the minutiae of research beyond writing and data reduction: researchers in the humanities value it less than those in pure and applied sciences; individuals from Far East Asia and Middle East and Northern Africa value these activities more than anglophones and northern Europeans. While developing national and international collaborations, researchers must recognize differences in peoples values while assigning authorship.


Perspectives on a mobile application that maps assistive technology resources in Africa.
Access to assistive technology (AT) is poor in African countries because of a lack of knowledge, resources, services and products. A mobile application (app), the AT-Info-Map, was developed to map AT availability in southern Africa. This article aimed to describe users' and suppliers' perceptions of the AT-Info-Map app. Qualitative data were collected in Zambia, Botswana, Malawi and Lesotho, through nine focus group discussions with 72 participants. Participants included AT users, AT suppliers and representatives of disability organisations. Data were thematically analysed. Two broad themes, that is, usefulness of the AT-Info-Map application and technical issues and content, emerged from the data analysis. Subthemes under usefulness focused on the importance of using current technology, convenience of the app, the need for accuracy, responsiveness of supplier to user's needs, influence on AT market and how the app creates an opportunity for networking. Challenges to download and navigate the app, the need for training in its use, exclusion of those not literate in English and those with visual impairments were subthemes under technical issues and content. The app was perceived as an important step to increase access to AT for persons with disabilities in less resourced settings. The challenges that emerged from the data analysis have led to the development of a web-based system that will complement or replace the app and improve AT information provision. However, the information provided by the app and website is still only a partial solution to improve AT access in Southern Africa.


Neuronal enriched endosomal protein of 21 kDa colocalizes with glutamate receptor subunit GLUR2/3 at the postsynaptic membrane.
Functional evidence suggests that neuronal enriched endosomal protein of 21 kDa (NEEP21) takes part in facilitating transport of AMPA receptors (AMPAR) in the synapse. To explore the anatomical basis for a role in this synaptic trafficking, we investigated the ultrastructural localization of NEEP21 in rodent brain. Using immunogold electron microscopy, we show that NEEP21 is colocalized with the AMPAR subunits GluR2/3 in postsynaptic spines. Quantitative analysis of gold particle distribution along an axis perpendicular to the postsynaptic specialization indicated that NEEP21 occurs in the postsynaptic membrane but also in the interior of the spines. NEEP21 positive endosomes/multivesicular bodies were found throughout cell bodies and dendrites. In light microscopical preparations, the NEEP21 antibody produced a labeling pattern in the neocortex, hippocampus and cerebellum that mimicked that of GluR2/3 and not that of GluR1 or 4. Our findings are consistent with a role for NEEP21 in facilitating vesicular transport of GluR2 between intracellular compartments and the postsynaptic plasma membrane.


A drop in the bucket of the megadiverse chewing louse genus Myrsidea (Phthiraptera, Amblycera, Menoponidae): ten new species from Amazonian Brazil.
Ten new species of Myrsidea Waterston, 1915 parasitic on members of the avian families Formicariidae, Thraupidae, Tyrannidae, Troglodytidae and Icteridae are described herein. They and their type hosts are M. isacantha sp. n. ex Chamaeza nobilis Gould, M. circumsternata sp. n. ex Formicarius colma Boddaert (Formicariidae); M. cacioppoi sp. n. ex Lanio fulvus (Boddaert), M. brasiliensis sp. n. ex Tangara chilensis (Vigors), M. saviti sp. n. ex Tangara schrankii (Spix) (Thraupidae), M. rodriguesae sp. n. ex Cnipodectes subbrunneus (Sclater), M. cnemotriccola sp. n. ex Cnemotriccus fuscatus (Wied-Neuwied), M. lathrotriccola sp. n. ex Lathrotriccus euleri (Cabanis) (Tyrannidae), M. faccioae sp. n. ex Cyphorhinus arada transfluvialis (Todd) (Troglodytidae), and M. lampropsaricola sp. n. ex Lampropsar tanagrinus (Spix) (Icteridae). Among these are two new Myrsidea species described from the avian family Formicariidae, which previously had only a single described Myrsidea species, and a new host record for M. cinnamomei Dalgleish et Price, 2005 ex Attila citriniventris Sclater. Analysis of mitochondrial cytochrome oxidase I sequences for these and other neotropical Myrsidea species provides an assessment of their phylogenetic relationships and indicates that all of these newly described species are genetically distinct. We also put these descriptions into context by estimating the potential number of unnamed Myrsidea species in Brazil, given the known diversity of potential hosts and typical levels of host specificity for Myrsidea species. Our estimate indicates that Brazilian Myrsidea species diversity is likely more than an order of magnitude greater than the number of described Myrsidea species known from Brazil, highlighting the need for future work on this megadiverse ectoparasite genus.


Mechanism for excitation-dependent photoluminescence from graphene quantum dots and other graphene oxide derivates: consensus, debates and challenges.
Luminescent nanomaterials, with wide applications in biosensing, bioimaging, illumination and display techniques, have been consistently garnering enormous research attention. In particular, those with wavelength-controllable emissions could be highly beneficial. Carbon nanostructures, including graphene quantum dots (GQDs) and other graphene oxide derivates (GODs), with excitation-dependent photoluminescence (PL), which means their fluorescence color could be tuned simply by changing the excitation wavelength, have attracted lots of interest. However the intrinsic mechanism for the excitation-dependent PL is still obscure and fiercely debated presently. In this review, we attempt to summarize the latest efforts to explore the mechanism, including the quantum confinement effect, surface traps model, giant red-edge effect, edge states model and electronegativity of heteroatom model, as well as the newly developed synergistic model, to seek some clues to unravel the mechanism. Meanwhile the controversial difficulties for each model are further discussed. Besides this, the challenges and potential influences of the synthetic methodology and development of the materials are illustrated extensively to elicit more thought and constructive attempts toward their application.


Construction and in vitro properties of SIVmac mutants with deletions in "nonessential" genes.
The so-called "nonessential" genes of primate lentiviruses can be deleted without abrogating the ability of virus to replicate under at least some cell culture conditions. In SIVmac, these genes are vif, vpx, vpr, and nef. Sequences in the upstream region of U3 in the LTR have also been shown to be dispensable for replication in cell culture. We report here the construction and characterization of a panel of 40 single and combination deletion mutants derived from the pathogenic molecular clone SIVmac239. Deletion of the vpr gene caused little or no change in the growth properties of SIVmac239 in CEMx174 cells, in rhesus monkey peripheral blood mononuclear cells (PBMCs), or in rhesus monkey alveolar macrophages. Deletion of the vpx gene resulted in a greatly reduced rate of replication of the virus in the primary PBMC and macrophage cultures, but no significant reduction in replication of the virus in CEMx174 cells. Deletion of the vpx gene appeared to have a greater effect on virus replication in macrophages than in PBMCs. Deletion of the vif gene caused a dramatic reduction in replication in all cell types tested. However, even delta 5, which contains deletions in all five targeted regions (vif, vpx, vpr, nef, and U3), can still replicate in CEMx174 cells albeit with greatly delayed kinetics. Deletion of nef, alone or in combination with deletions in U3 and vpr, had no observable effect on replication of the virus in any of the cells tested. Because the disease induced by the parental SIVmac239 clone in rhesus monkeys has been well characterized and is remarkably similar to AIDS in humans, this collection of mutants will be useful for relating in vitro properties and gene function with in vivo pathogenic potential.


Colposcopy Standards: Guidelines for Endocervical Curettage at Colposcopy.
The most recent guidelines for colposcopy practice in the United States, the 2017 Colposcopy Standards Consensus Guidelines, did not include recommendations for endocervical curettage (ECC). This document provides updated guidelines for use of ECC among patients referred for colposcopy. Consensus guidelines for the use of ECC were developed in 2012. To update these guidelines in concordance with the 2017 Colposcopy Standards process, an expert workgroup was convened in 2021. Literature had been previously reviewed through 2011, before the 2012 guideline. Literature from the years 2012-2021 and data from the NCI Biopsy study were reviewed, focusing on the additional yield of ECC. Endocervical curettage is recommended for patients with high-grade cytology, human papillomavirus 16/18 infection, positive results on dual staining for p16/Ki67, for those previously treated for known or suspected cervical precancer or considering observation of cervical intraepithelial neoplasia grade 2, and when the squamocolumnar junction is not fully visualized at colposcopy. Endocervical curettage is preferred for all patients aged older than 40 years. Endocervical curettage is acceptable for all nonpregnant patients undergoing colposcopy but may be omitted when a subsequent excisional procedure is planned, the endocervical canal does not admit a sampling device, or in nulliparous patients aged younger than 30 years, with cytology reported as atypical squamous cells of undetermined significance or low-grade squamous intraepithelial lesion regardless of whether the squamocolumnar junction is fully visualized. Endocervical curettage is unacceptable in pregnancy. These guidelines for ECC add to the 2017 consensus recommendations for colposcopy practice in the United States.


Hedgehog and Wingless stabilize but do not induce cell fate during Drosophila dorsal embryonic epidermal patterning.
A fundamental concept in development is that secreted molecules such as Wingless (Wg) and Hedgehog (Hh) generate pattern by inducing cell fate. By following markers of cellular identity posterior to the Wg- and Hh-expressing cells in the Drosophila dorsal embryonic epidermis, we provide evidence that neither Wg nor Hh specifies the identity of the cell types they pattern. Rather, they maintain pre-existing cellular identities that are otherwise unstable and progress stepwise towards a default fate. Wg and Hh therefore generate pattern by inhibiting specific switches in cell identity, showing that the specification and the patterning of a given cell are uncoupled. Sequential binary decisions without induction of cell identity give rise to both the groove cells and their posterior neighbors. The combination of independent progression of cell identity and arrest of progression by signals facilitates accurate patterning of an extremely plastic developing epidermis.


All the disorder mechanisms in the 13:58 phases come together. Out of the modulated confusion rises the remarkable phase Ce12.60Cd58.68(2).
The compound Ce12.60Cd58.68(2) is a metrically commensurate representative of the incommensurately modulated phase Ce13Cd57+delta. It is most likely a lock-in phase. The structure, which was solved using seeding of the modulation from those positions most affected as well as direct solution by charge flipping, represents a rare case of ordering in a family of structures where disorder is the rule. The disorder mechanisms, known from other RE13Zn/Cd58 phases, order in this phase to give rise to a remarkable interplay between interstitials and vacancies. The compound crystallizes in the super space group Ccmm(alpha00)00s (standard setting Amma(00gamma)s00) with cell parameters a=27.2789(14) A, b=15.7592(1) A, and c=15.5816(4) A and a q vector close to 2/3.


Using patient-reported measurement to pave the path towards personalized medicine.
Given the potential and importance of personalized or individualized medicine for health care delivery and its effects on patients' quality of life, a plenary session was devoted to personalized medicine during the 19th Annual Conference of the International Society for Quality of Life Research held in October 2012 in Budapest, Hungary. This paper summarizes the three presentations and discusses their implications for quality-of-life research. Reviews of the literature and presentation of empirical studies. Personalized screening for breast cancer. To individualize screening and only target those women with an increased risk for breast cancer, researchers at the Karolinska Institutet in Stockholm perform a large population-based study to identify high-risk women based on lifestyle, genetics, mammographic morphology, and other markers as well as quality of life. Personalized support for treatment adherence. Inclusion of a simple, brief adherence measure into the clinical visit has demonstrated significant improvement in medication-taking behaviour and resultant improvement in health status. Personalized diagnosis of mental disorders. The DSM-5, the current manual for mental disorders, contains patient-based symptom and diagnosis severity measures that allow more individualized diagnosis than was hitherto possible. Personalized medicine will continue to be increasingly applied and holds the potential to improve health outcomes including quality of life. At the same time, it will invite a host of new ethical, practical, and psychosocial questions. Further reflection and discussion of how our field can embrace and address these emerging challenges is needed.


Apoptosis of nur77/N10-transgenic thymocytes involves the Fas/Fas ligand pathway.
The orphan nuclear receptor Nur77/N10 has recently been demonstrated to be involved in apoptosis of T cell hybridomas. We report here that chronic expression of Nur77/N10 in thymocytes of transgenic mice results in a dramatic reduction of CD4+CD8+ double-positive as well as CD4+CD8- and CD4-CD8+ single-positive cell populations due to an early onset of apoptosis. CD4-CD8- double-negative and CD25+ precursor cells, however, are unaffected. Moreover, nur77/N10-transgenic thymocytes show increased expression of Fas ligand (FasL), while the levels of the Fas receptor (Fas) are not increased. The mouse spontaneous mutant gld (generalized lymphoproliferative disease) carries a point mutation in the extracellular domain of the FasL gene that abolishes the ability of FasL to bind to Fas. Thymuses from nur77/N10-transgenic mice on a gld/gld background have increased cellularity and an almost normal profile of thymocyte subpopulations. Our results demonstrate that one pathway of apoptosis triggered by Nur77/N10 in double-positive thymocytes occurs through the upregulation of FasL expression resulting in increased signaling through Fas.


Identification of a Streptococcus sanguis receptor for salivary agglutinins.
The objective of this study was to characterize a fraction from oral streptococci containing receptor activity for salivary agglutinin molecules. Several species and strains of streptococci were disrupted in a Ribi press. The supernatant was nuclease-treated and subjected to differential centrifugation. Receptor activity in the fractions was measured by the inhibition of saliva-mediated bacterial aggregation. In addition, bacterial strains were tested for their ability to aggregate and to deplete saliva of agglutinin activity. Three patterns of activity were observed: Streptococcus sanguis M5 depleted saliva of agglutinin activity and aggregated well; Streptococcus sanguis CC5A depleted saliva of agglutinin but did not aggregate well; and Streptococcus faecalis S-161 neither depleted saliva of agglutinin nor did it aggregate. The 105,000 g supernatant fractions derived from Ribi-disrupted Streptococcus sanguis M5 and CC5A, but not from Streptococcus faecalis, showed dose-dependent inhibition of saliva-mediated aggregation. This inhibitory activity was non-dialyzable, had the same heat and trypsin sensitivity as that seen with intact bacteria, and was not due to enzymatic digestion of the salivary agglutinin. Iso-electric focusing revealed a single active region with a pI of 5.5 which was clearly separated from the bulk of the bacterial proteins.


Light extinction by fine atmospheric particles in the White Mountains region of New Hampshire and its relationship to air mass transport.
Chemical, optical, and physical measurements of fine aerosols (aerodynamic diameter < or = 2.5 microm) have been performed at a mountaintop location adjacent to the White Mountain National Forest in northern NH, USA. A 1-month long sampling campaign was conducted at Cranmore Mountain during spring 2000. We report on the apportionment of light extinction by fine aerosols into its major chemical components, and relationships between variations in aerosol parameters and changes in air mass origin. Filter-based, 24-h integrated samples were collected and analyzed for major inorganic ions, as well as organic (OC), elemental (EC), and total carbon. Light scattering and light absorption coefficients were measured at 5-min intervals using an integrating nephelometer and a light absorption photometer. Fine particle number density was measured with a condensation particle counter. Air mass origins and transport patterns were investigated through the use of 3-day backward trajectories and a synoptic climate classification system. Two distinct transport regimes were observed: (1) flow from the north/northeast (N/NE) occurred during 9 out of 18 sample-days; and (2) flow from the west/southwest (W/SW) occurred 8 out of 18 sample-days. All measured and derived aerosol and meteorological parameters were separated into two categories based on these different flow scenarios. During W/SW flow, higher values of aerosol chemical concentration, absorption and scattering coefficients, number density, and haziness were observed compared to N/NE flow. The highest level of haziness was associated with the climate classification Frontal Atlantic Return, which brought polluted air into the region from the mid-Atlantic corridor. Fine particle mass scattering efficiencies of (NH4)2SO4 and OC were 5.35 +/- 0.42 m2 g(-1) and 1.56 +/- 0.40 m2 g(-1), respectively, when transport was out of the N/NE. When transport was from the W/SW the values were 4.94 +/- 0.68 m2 g(-1) for (NH4)2SO4 and 2.18 +/- 0.91 m2 g(-1) for OC. EC mass absorption efficiency when transport was from the N/NE was 9.66 +/- 1.06 m2 g(-1) and 10.80 +/- 1.76 m2 g(-1) when transport was from the W/SW. Results from this work can be used to predict visual air quality in the White Mountain National Forest based on a forecasted synoptic climate classification and its associated visibility.


Characterisation of Deubiquitylating Enzymes in the Cellular Response to High-LET Ionizing Radiation and Complex DNA Damage.
Ionizing radiation, particular high-linear energy transfer (LET) radiation, can induce complex DNA damage (CDD) wherein 2 or more DNA lesions are induced in close proximity, which contributes significantly to the cell killing effects. However, knowledge of the enzymes and mechanisms involved in coordinating the recognition and processing of CDD in cellular DNA are currently lacking. A small interfering RNA screen of deubiquitylation enzymes was conducted in HeLa cells irradiated with high-LET α-particles or protons, versus low-LET protons and x-rays, and cell survival was monitored by clonogenic assays. Candidates whose depletion led to decreased cell survival specifically in response to high-LET radiation were validated in both HeLa and oropharyngeal squamous cell carcinoma (UMSCC74A) cells, and the association with CDD repair was confirmed using an enzyme modified neutral comet assay. Depletion of USP6 decreased cell survival specifically after high-LET α-particles and protons, but not low-LET protons or x-rays. USP6 depletion caused cell cycle arrest and a deficiency in CDD repair mediated through instability of poly(ADP-ribose) polymerase-1 (PARP-1) protein. Increased radiosensitivity of cells to high-LET protons as a consequence of defective CDD repair was furthermore mimicked using the PARP inhibitor olaparib, and through PARP-1 small interfering RNA. USP6 controls cell survival in response to high-LET radiation by stabilizing PARP-1 protein levels, which is essential for CDD repair. We also describe synergy between CDD induced by high-LET protons and PARP inhibition, or PARP-1 depletion, in effective cancer cell killing.


Efficient first-principles calculation of the quantum kinetic energy and momentum distribution of nuclei.
Light nuclei at room temperature and below exhibit a kinetic energy which significantly deviates from the predictions of classical statistical mechanics. This quantum kinetic energy is responsible for a wide variety of isotope effects of interest in fields ranging from chemistry to climatology. It also furnishes the second moment of the nuclear momentum distribution, which contains subtle information about the chemical environment and has recently become accessible to deep inelastic neutron scattering experiments. Here, we show how, by combining imaginary time path integral dynamics with a carefully designed generalized Langevin equation, it is possible to dramatically reduce the expense of computing the quantum kinetic energy. We also introduce a transient anisotropic Gaussian approximation to the nuclear momentum distribution which can be calculated with negligible additional effort. As an example, we evaluate the structural properties, the quantum kinetic energy, and the nuclear momentum distribution for a first-principles simulation of liquid water.


Layers of listening: qualitative analysis of the impact of early intervention services for first-episode psychosis on carers' experiences.
Early intervention services (EIS) comprise low-stigma, youth-friendly mental health teams for young people undergoing first-episode psychosis (FEP). Engaging with the family of the young person is central to EIS policy and practice.AimsBy analysing carers' accounts of their daily lives and affective challenges during a relative's FEP against the background of wider research into EIS, this paper explores relationships between carers' experiences and EIS. Semi-structured longitudinal interviews with 80 carers of young people with FEP treated through English EIS. Our data suggest that EIS successfully aid carers to support their relatives, particularly through the provision of knowledge about psychosis and medications. However, paradoxical ramifications of these user-focused engagements also emerge; they risk leaving carers' emotions unacknowledged and compounding an existing lack of help-seeking. By focusing on EIS's engagements with carers, this paper draws attention to an urgent broader question: as a continuing emphasis on care outside the clinic space places family members at the heart of the care of those with severe mental illness, we ask: who can, and should, support carers, and in what ways?


Evolutionary Dynamics of Male Reproductive Genes in the Drosophila virilis Subgroup.
Postcopulatory sexual selection (PCSS) is a potent evolutionary force that can drive rapid changes of reproductive genes within species, and thus has the potential to generate reproductive incompatibilities between species. Male seminal fluid proteins (SFPs) are major players in postmating interactions, and are important targets of PCSS in males. The virilis subgroup of Drosophila exhibits strong interspecific gametic incompatibilities, and can serve as a model to study the genetic basis of PCSS and gametic isolation. However, reproductive genes in this group have not been characterized. Here we utilize short-read RNA sequencing of male reproductive organs to examine the evolutionary dynamics of reproductive genes in members of the virilis subgroup: D. americana, D. lummei, D. novamexicana, and D. virilis We find that the majority of male reproductive transcripts are testes-biased, accounting for ∼15% of all annotated genes. Ejaculatory bulb (EB)-biased transcripts largely code for lipid metabolic enzymes, and contain orthologs of the D. melanogaster EB protein, Peb-me, which is involved in mating-plug formation. In addition, we identify 71 candidate SFPs, and show that this gene set has the highest rate of nonsynonymous codon substitution relative to testes- and EB-biased genes. Furthermore, we identify orthologs of 35 D. melanogaster SFPs that have conserved accessory gland expression in the virilis group. Finally, we show that several of the SFPs that have the highest rate of nonsynonymous codon substitution reside on chromosomal regions, which contributes to paternal gametic incompatibility between species. Our results show that SFPs rapidly diversify in the virilis group, and suggest that they likely play a role in PCSS and/or gametic isolation.


Untangling glycaemia and mortality in critical care.
Hyperglycaemia is associated with adverse outcomes in the intensive care unit, and initial studies suggested outcome benefits of glycaemic control (GC). However, subsequent studies often failed to replicate these results, and they were often unable to achieve consistent, safe control, raising questions about the benefit or harm of GC as well as the nature of the association of glycaemia with mortality and clinical outcomes. In this study, we evaluated if non-survivors are harder to control than survivors and determined if glycaemic outcome is a function of patient condition and eventual outcome or of the glycaemic control provided. Clinically validated, model-based, hour-to-hour insulin sensitivity (SI) and its hour-to-hour variability (%ΔSI) were identified over the first 72 h of therapy in 145 patients (119 survivors, 26 non-survivors). In hypothesis testing, we compared distributions of SI and %ΔSI in 6-hourly blocks for survivors and non-survivors. In equivalence testing, we assessed if differences in these distributions, based on blood glucose measurement error, were clinically significant. SI level was never equivalent between survivors and non-survivors (95% CI of percentage difference in medians outside ±12%). Non-survivors had higher SI, ranging from 9% to 47% higher overall in 6-h blocks, and this difference became statistically significant as glycaemic control progressed. %ΔSI was equivalent between survivors and non-survivors for all 6-hourly blocks (95% CI of difference in medians within ±12%) and decreased in general over time as glycaemic control progressed. Whereas non-survivors had higher SI levels, variability was equivalent to that of survivors over the first 72 h. These results indicate survivors and non-survivors are equally controllable, given an effective glycaemic control protocol, suggesting that glycaemia level and variability, and thus the association between glycaemia and outcome, are essentially determined by the control provided rather than by underlying patient or metabolic condition.


Osteopontin is not required for the development of Th1 responses and viral immunity.
Osteopontin (OPN) has been defined as a key cytokine promoting the release of IL-12 and hence inducing the development of protective cell-mediated immunity to viruses and intracellular pathogens. To further characterize the role of OPN in antiviral immunity, OPN-deficient (OPN-/-) mice were analyzed after infection with influenza virus and vaccinia virus. Surprisingly, we found that viral clearance, lung inflammation, and recruitment of effector T cells to the lung were unaffected in OPN-/- mice after influenza infection. Furthermore, effector status of T cells was normal as demonstrated by normal IFN-gamma production and CTL lytic activity. Moreover, activation and Th1 differentiation of naive TCR transgenic CD4+ T cells by dendritic cells and cognate Ag was normal in the absence of OPN in vitro. Contrary to a previous report, we found that OPN-/- mice mounted a normal immune response to Listeria monocytogenes. In conclusion, OPN is dispensable for antiviral immune responses against influenza virus and vaccinia virus.


Natural killer cell inhibits human immunodeficiency virus replication in chronically infected immune cells.
Natural killer (NK) cells are a crucial component of the host innate immune system. We investigated the noncytolytic anti-human immunodeficiency virus (HIV) activity of NK cells in chronically HIV-infected immune cells. Supernatants collected from NK cell cultures (both primary NK cells and NK cell lines, YTS and NK 92) inhibited HIV activation in peripheral blood mononuclear cells (PBMCs) from HIV-infected subjects. NK supernatants (NK SN) also suppressed tumor necrosis factor (TNF)-alpha-induced HIV activation in chronically infected cell lines (U1 and ACH-2 cells). The antibody to interferon (IFN)-gamma blocked NK SN-mediated anti-HIV effect, while the antibodies to CC-chemokines had no impact on NK SN-mediated HIV inhibition in U1 and ACH-2 cells. Investigation of mechanism(s) responsible for the NK action showed that NK SN inhibited TNF-alpha-mediated activation of HIV-long-terminal repeat (LTR), and upregulated the expression of signal transducer and activator of transcription (STAT)-1 and phosphorylated P38 mitogen-activated protein kinase (MAPK). The P38 MAPK inhibitor (SB 203580) blocked NK SN-mediated HIV inhibition. These data provide compelling evidence that NK cells have a critical role in controlling HIV activation in the reservoirs.


Cancer Cells with Alternative Lengthening of Telomeres Do Not Display a General Hypersensitivity to ATR Inhibition.
Telomere maintenance is a hallmark of cancer as it provides cancer cells with cellular immortality. A significant fraction of tumors uses the alternative lengthening of telomeres (ALT) pathway to elongate their telomeres and to gain an unlimited proliferation potential. Since the ALT pathway is unique to cancer cells, it represents a potentially valuable, currently unexploited target for anti-cancer therapies. Recently, it was proposed that ALT renders cells hypersensitive to ataxia telangiectasia- and RAD3-related (ATR) protein inhibitors (Flynn et al., Science 347, 273). Here, we measured the response of various ALT- or telomerase-positive cell lines to the ATR inhibitor VE-821. In addition, we compared the effect of the inhibitor on cell viability in isogenic cell lines, in which ALT was active or suppressed. In these experiments, a general ATR inhibitor sensitivity of cells with ALT could not be confirmed. We rather propose that the observed variations in sensitivity reflect differences between cell lines that are unrelated to ALT.