Care of patients with chronic disease: the challenge for general practice.
General practice can provide good quality care for a range of high-prevalence chronic diseases, at the same time providing continuity of care and management of comorbidity. Although the quality of care for patients with chronic disease is improving in general practice, about half of patient care does not meet optimal standards. Factors contributing to the gap between optimal and current practice include the method of financing, the availability of other disciplines to participate in team care, limited engagement with self-management education, and lack of information and decision support systems. National initiatives and incentives have enhanced planned and systematic care in general practice, and some programs have been introduced to improve access to allied health care. The number and complexity of programs, and lack of integration between them are a significant administrative burden for general practice, and the financial incentives are small compared to overseas programs. A better integrated and more comprehensive strategy is required to achieve widespread and sustained improvements in the quality of care for people with chronic disease in general practice.


The Relative Association of Collective Efficacy in School and Neighborhood Contexts With Adolescent Alcohol Use.
It is unclear whether either neighborhood collective efficacy or school collective efficacy is associated with adolescent alcohol use. This study aimed to examine the relative contributions of collective efficacy, both in school and in the neighborhood contexts, to alcohol use among Japanese adolescents. A cross-sectional study was conducted in public high schools across Okinawa and Ibaraki Prefectures in Japan in 2016. The study participants consisted of 3,291 students in grades 10 through 12 cross-nested in 51 schools and 107 neighborhoods. Alcohol use was measured as current alcohol drinking, which was defined as self-reported drinking on at least 1 day in the past 30 days. Collective efficacy was measured using scales of social cohesion and informal social control in school and the neighborhood. Contextual-level collective efficacy was measured using aggregated school-level and neighborhood-level individual responses, respectively. We used non-hierarchical multilevel models to fit the cross-nested data. Significant variation in alcohol use was shown between schools but not between neighborhoods. After adjusting for covariates, school collective efficacy at individual- and contextual-levels was protectively associated with alcohol drinking (odds ratio [OR] for the increase of one standard deviation from the mean 0.72; 95% confidence interval [CI], 0.63-0.82 and OR 0.61; 95% CI, 0.49-0.75, respectively), whereas neighborhood collective efficacy at individual- and contextual-levels was not associated with alcohol consumption. The school-level associations of collective efficacy with adolescent alcohol use may have the greater impact than the neighborhood-level associations. Adolescent drinking prevention efforts should include enhancing school collective efficacy.


Mutations in four glycosyl hydrolases reveal a highly coordinated pathway for rhodopsin biosynthesis and N-glycan trimming in Drosophila melanogaster.
As newly synthesized glycoproteins move through the secretory pathway, the asparagine-linked glycan (N-glycan) undergoes extensive modifications involving the sequential removal and addition of sugar residues. These modifications are critical for the proper assembly, quality control and transport of glycoproteins during biosynthesis. The importance of N-glycosylation is illustrated by a growing list of diseases that result from defects in the biosynthesis and processing of N-linked glycans. The major rhodopsin in Drosophila melanogaster photoreceptors, Rh1, is highly unique among glycoproteins, as the N-glycan appears to be completely removed during Rh1 biosynthesis and maturation. However, much of the deglycosylation pathway for Rh1 remains unknown. To elucidate the key steps in Rh1 deglycosylation in vivo, we characterized mutant alleles of four Drosophila glycosyl hydrolases, namely α-mannosidase-II (α-Man-II), α-mannosidase-IIb (α-Man-IIb), a β-N-acetylglucosaminidase called fused lobes (Fdl), and hexosaminidase 1 (Hexo1). We have demonstrated that these four enzymes play essential and unique roles in a highly coordinated pathway for oligosaccharide trimming during Rh1 biosynthesis. Our results reveal that α-Man-II and α-Man-IIb are not isozymes like their mammalian counterparts, but rather function at distinct stages in Rh1 maturation. Also of significance, our results indicate that Hexo1 has a biosynthetic role in N-glycan processing during Rh1 maturation. This is unexpected given that in humans, the hexosaminidases are typically lysosomal enzymes involved in N-glycan catabolism with no known roles in protein biosynthesis. Here, we present a genetic dissection of glycoprotein processing in Drosophila and unveil key steps in N-glycan trimming during Rh1 biosynthesis. Taken together, our results provide fundamental advances towards understanding the complex and highly regulated pathway of N-glycosylation in vivo and reveal novel insights into the functions of glycosyl hydrolases in the secretory pathway.


Sexual satisfaction assessment in 194 nonmetastatic cancer patients on treatment or in follow-up.
Sexuality is an important aspect of quality of life, but health care professionals still avoid discussing sexual issues with cancer patients. We present a secondary analysis of sexuality issues according to the results of a survey on 266 patients with early-stage cancer. The aim of the survey was to ascertain the feasibility and clinical usefulness of questionnaires (Patient Dignity Inventory, PDI; Hospital Anxiety and Depression Scale, HADS; Edmonton Symptom Assesssment Scale, ESAS; FACIT-spiritual well-being scale, FACIT-SP; System of Belief Inventory, SBI-15R) investigating aspects such as dignity, hope and research of meaning in life. The present study is an ancillary analysis of the full sample, and we have focused on the results of FACIT-SP about the correlation between sexual satisfaction and clinical characteristics in 108 patients having solid tumors and 86 having hematological malignancies with no metastases who were on active cancer treatment or in follow-up in four different cancer treatment settings during the first half of 2011. The median age of the 194 patients was 65 years, 112 were women, 155 were undergoing treatment and 39 were in follow-up. Eighty-three patients were above the cutoff score for HADS. Among the 171 believers, 80 were churchgoers and 91 were nonchurchgoers, whereas the nonbelievers among the patients were 23. Thirty-five percent of the patients did not respond to the sexuality item of the questionnaire. Among the responders (n = 126), 36% reported having no sexual satisfaction (score = 0). Sexual dissatisfaction was greater in older patients (47% vs 31%, not significant [NS]), women (43% vs 27%, NS), patients on treatment (38% vs 25%, NS), patients who requested psychological support (53% vs 25%, P = 0.001), patients with high levels of anxiety and depression, i.e., HADS scores >10 (44% vs 30%, NS), nonbelievers (61% vs 34% among churchgoers, 29% among believers but nonchurchgoers, P = 0.046). One out of 3 patients did not respond to the item on sexuality. Among the responders, 1 out of 3 reported having no sexual satisfaction. Half of the patients receiving psychological support considered their sexual life not satisfying. Clinical interviews and specific questionnaires on sexuality should be used to investigate this particular aspect.


Prophylactic use of melatonin protects against focal cerebral ischemia in mice: role of endothelin converting enzyme-1.
Melatonin has previously been shown to be neuroprotective in rodent models of ischemic stroke. Herein, we tested whether this antioxidant may also be suitable for prophylactic use against stroke. To clarify this issue, melatonin was administrated orally for 9 wk (4 mg/kg/day) in mice and its effects on subsequent injury development after 90 min of intraluminal middle cerebral artery (MCA) occlusion were tested. To evaluate its neuroprotective properties, the protective actions of prophylactic melatonin were compared with both acute melatonin (4 mg/kg, i.p.) administration and with a diluent (sham)-treated control condition. MCA occlusion resulted in reproducible ischemia, as revealed by laser Doppler flowmetry; this was followed by a rapid restoration of blood flow immediately after reperfusion onset. Laser Doppler flow values after reperfusion onset were moderately elevated by melatonin, both when the indole was given prophylactically and when acutely administrated after stroke. In control animals, reproducible brain infarcts were observed 24 hr after reperfusion onset. Treatment with melatonin significantly reduced the infarct size by approximately 30-35%, independent of whether the indole was given prophylactically before or acutely after ischemia. To test whether brain protection involved vascular mechanisms, as suggested earlier, the effects of melatonin on endothelin converting enzyme-1 (ECE-1) levels were studied using Western blots. Interestingly, delivery of melatonin was accompanied by a marked inhibition of ECE-1 levels, which was similarly seen after both acute and chronic melatonin treatment. Our data suggest that melatonin, given at pharmacological doses, may be suitable as a prophylaxis against stroke. Tissue protection may involve an inhibition of ECE-1, which improves vasodilation, after ischemia.


A human skin equivalent burn model to study the effect of a nanocrystalline silver dressing on wound healing.
In this study, a deep burn wound model was established using a 3D human skin equivalent (HSE) model and this was compared to native skin. HSEs were constructed from dermis derived from abdominoplasty/breast surgery and this dermal template was seeded with primary keratinocytes and fibroblasts. The HSE model was structurally similar to native skin with a stratified and differentiated epidermis. A contact burn (60 °C, 80 °C, 90 °C) was applied with a modified soldering iron and wounds were observed at day 1 and 7 after burn. The HSEs demonstrated re-growth with keratinocyte proliferation and formation of a neo-epidermis after burn injury, whereas the ex vivo native skin did not. To assess the suitability of the 3D HSE model for penetration and toxicity studies, a nanocrystalline silver dressing was applied to the model for 7 days, with and without burn injury. The effect of silver on skin re-growth and its penetration and subcellular localization was assessed in HSEs histologically and with laser ablation-inductively coupled plasma mass spectrometry (LA-ICPMS). The silver treatment delayed or reduced skin re-growth, and silver particles were detected on the top of the epidermis, and within the papillary dermis. This novel in vitro 3D multicellular deep burn wound model is effective for studying the pathology and treatment of burn wound injury and is suitable for penetration and toxicity studies of wound healing treatments.


Tolerance to multiple climate stressors: a case study of Douglas-fir drought and cold hardiness.
Drought and freeze events are two of the most common forms of climate extremes which result in tree damage or death, and the frequency and intensity of both stressors may increase with climate change. Few studies have examined natural covariation in stress tolerance traits to cope with multiple stressors among wild plant populations.We assessed the capacity of coastal Douglas-fir (Pseudotsuga menziesii var. menziesii), an ecologically and economically important species in the northwestern USA, to tolerate both drought and cold stress on 35 populations grown in common gardens. We used principal components analysis to combine drought and cold hardiness trait data into generalized stress hardiness traits to model geographic variation in hardiness as a function of climate across the Douglas-fir range.Drought and cold hardiness converged among populations along winter temperature gradients and diverged along summer precipitation gradients. Populations originating in regions with cold winters had relatively high tolerance to both drought and cold stress, which is likely due to overlapping adaptations for coping with winter desiccation. Populations from regions with dry summers had increased drought hardiness but reduced cold hardiness, suggesting a trade-off in tolerance mechanisms.Our findings highlight the necessity to look beyond bivariate trait-climate relationships and instead consider multiple traits and climate variables to effectively model and manage for the impacts of climate change on widespread species.


High-intensity Raf signal causes cell cycle arrest mediated by p21Cip1.
Activated Raf has been linked to such opposing cellular responses as the induction of DNA synthesis and the inhibition of proliferation. However, it remains unclear how such a switch in signal specificity is regulated. We have addressed this question with a regulatable Raf-androgen receptor fusion protein in murine fibroblasts. We show that Raf can cause a G1-specific cell cycle arrest through induction of p21Cip1. This in turn leads to inhibition of cyclin D- and cyclin E-dependent kinases and an accumulation of hypophosphorylated Rb. Importantly, this behavior can be observed only in response to a strong Raf signal. In contrast, moderate Raf activity induces DNA synthesis and is sufficient to induce cyclin D expression. Therefore, Raf signal specificity can be determined by modulation of signal strength presumably through the induction of distinct protein expression patterns. Similar to induction of Raf, a strong induction of activated Ras via a tetracycline-dependent promoter also causes inhibition of proliferation and p21Cip1 induction at high expression levels. Thus, p21Cip1 plays a key role in determining cellular responses to Ras and Raf signalling. As predicted by this finding we show that Ras and loss of p21 cooperate to confer a proliferative advantage to mouse embryo fibroblasts.


De novo missense variants in ZBTB47 are associated with developmental delays, hypotonia, seizures, gait abnormalities, and variable movement abnormalities.
The collection of known genetic etiologies of neurodevelopmental disorders continues to increase, including several syndromes associated with defects in zinc finger protein transcription factors (ZNFs) that vary in clinical severity from mild learning disabilities and developmental delay to refractory seizures and severe autism spectrum disorder. Here we describe a new neurodevelopmental disorder associated with variants in ZBTB47 (also known as ZNF651), which encodes zinc finger and BTB domain-containing protein 47. Exome sequencing (ES) was performed for five unrelated patients with neurodevelopmental disorders. All five patients are heterozygous for a de novo missense variant in ZBTB47, with p.(Glu680Gly) (c.2039A>G) detected in one patient and p.(Glu477Lys) (c.1429G>A) identified in the other four patients. Both variants impact conserved amino acid residues. Bioinformatic analysis of each variant is consistent with pathogenicity. We present five unrelated patients with de novo missense variants in ZBTB47 and a phenotype characterized by developmental delay with intellectual disability, seizures, hypotonia, gait abnormalities, and variable movement abnormalities. We propose that these variants in ZBTB47 are the basis of a new neurodevelopmental disorder.


Multimodal imaging of the sonic organ of Porichthys notatus, the singing midshipman fish.
The sonic midshipman fish, Porichthys notatus, is a bottom-dwelling species whose swim bladder has evolved into a highly specialized, sound-producing organ. The males of this species exist in two distinct morphs with different physical characteristics and sexual strategies. The Type I males have a much larger sound organ and are capable of generating a loud approximately 100 Hz tone continuously for over an hour to attract females. This sound is produced by sonic muscle and represents one of the most superfast and super-enduring striated muscles found in nature. Each fiber contains a hollow, tubular contractile apparatus composed of radially arranged myofibrils with extremely broad Z-bands that are supported by a desmin-rich cytoskeleton. We have used micro computed tomography (CT) imaging and magnetic resonance (MR) imaging to visualize the location of the sonic organ in an intact male fish. We have also obtained high-resolution MR images of the excised swim bladders from both male types. The images of the Type I sonic organ are strikingly detailed and high-contrast, revealing both the internal organization of the bladder and the crisscrossing muscle fibers and their mode of attachment to the underlying bladder. The high-contrast variation in these images is due to different T(2) values for fiber bundles and the spaces between the bundles. Direct MR imaging of intact Type I sonic organ in Type I midshipman fish is a powerful approach to understanding the contraction of this superfast muscle and the oscillation of its bladder to produce mating calls, and how placement of the sonic organ in the body of the fish sheds light on its prodigious ability to produce and transmit its loud mating call.


Delivering a selective neonatal BCG vaccination programme in a multi-ethnic community: an audit of the neonatal BCG immunisation programme in Birmingham and Solihull.
Neonatal bacillus Calmette-Guérin (BCG) immunisation is a primary preventive measure against tuberculosis. Local health professionals expressed concern about the variability of knowledge regarding eligible infants and uptake of the vaccine. A questionnaire was sent out to health visitors for use at the routine visit to babies. Details requested included ethnic group and country of origin of the infant, eligibility for BCG vaccination, and vaccination status. BCG vaccination was indicated for 41% of newborns. In total 74% of these eligible infants received the vaccine with a range of 36-83% between the five maternity units. There were inconsistencies within maternity units in identifying high-risk groups by ethnicity and country of origin, resulting in low coverage in certain eligible groups. Confusion exists about which infants are at risk of tuberculosis and should be vaccinated. Current national guidelines are not specific enough for cases of interracial parenting and for the increasingly diverse countries of origin of the population. In the absence of clearer national guidelines there is a need for pragmatic local guidance.


A GIS-Based Artificial Neural Network Model for Spatial Distribution of Tuberculosis across the Continental United States.
Despite the usefulness of artificial neural networks (ANNs) in the study of various complex problems, ANNs have not been applied for modeling the geographic distribution of tuberculosis (TB) in the US. Likewise, ecological level researches on TB incidence rate at the national level are inadequate for epidemiologic inferences. We collected 278 exploratory variables including environmental and a broad range of socio-economic features for modeling the disease across the continental US. The spatial pattern of the disease distribution was statistically evaluated using the global Moran's I, Getis⁻Ord General G, and local Gi* statistics. Next, we investigated the applicability of multilayer perceptron (MLP) ANN for predicting the disease incidence. To avoid overfitting, L1 regularization was used before developing the models. Predictive performance of the MLP was compared with linear regression for test dataset using root mean square error, mean absolute error, and correlations between model output and ground truth. Results of clustering analysis showed that there is a significant spatial clustering of smoothed TB incidence rate (p < 0.05) and the hotspots were mainly located in the southern and southeastern parts of the country. Among the developed models, single hidden layer MLP had the best test accuracy. Sensitivity analysis of the MLP model showed that immigrant population (proportion), underserved segments of the population, and minimum temperature were among the factors with the strongest contributions. The findings of this study can provide useful insight to health authorities on prioritizing resource allocation to risk-prone areas.


Quantitative computed tomography liver perfusion imaging using dynamic spiral scanning with variable pitch: feasibility and initial results in patients with cancer metastases.
To assess the feasibility and image quality of computed tomography (CT) liver perfusion imaging using an adaptive 4D spiral-mode, developed to extend the z-axis coverage, and to report initial qualitative and quantitative results in patients with cancer metastases. A total of 21 patients with liver metastases of various origins underwent CT perfusion imaging (100 kV and 150 mAs/rot) using a 4D spiral-mode with single-source 64-slice CT (n = 7) with a scan range of 6.7 cm (protocol A: 16 cycles, 46.5 seconds examination time), or dual-source 128-slice CT with a scan range of 14.8 cm (protocol B: 16 cycles, 46.5 seconds examination time, n = 7; protocol C: 12 cycles, 51.0 seconds examination time, n = 7). Ability to suspend respiration during perfusion imaging was monitored. Two independent readers assessed image quality on a 4-point scale, both before and after motion correction, and performed a qualitative (ie, arterial enhancement pattern and enhancement change over time) and quantitative perfusion (ie, arterial liver perfusion [ALP]; portal-venous perfusion [PVP]; hepatic perfusion index [HPI]) analysis. Of 21 patients, 7 (33%) could suspend respiration throughout the perfusion study and 14 (67%) resumed shallow breathing during the perfusion scan. The 21 patients had a total of 88 metastases. The scan range of protocol A covered at least 1 metastasis in all patients (total 20/34 [58.8%] metastases). The scan range of protocol B and C covered 53 of 54 (98.1%) metastases, whereas one metastasis in segment VIII was only partially imaged. Image quality was diagnostic both before and after motion correction, whereas being significantly better after motion correction (P < 0.001). Qualitative perfusion analysis of 67 metastases revealed diffuse arterial enhancement in 3 (4.5%), sparse enhancement in 11 (16.4%), peripheral-nodular enhancement in 9 (13.4%), rim-like enhancement in 15 (22.4%), and none in 29 (43.3%) metastases. Enhancement over time of 67 metastases showed a centripetal progression in 6 (8.9%), sustained portal phase in 16 (23.9%), wash-out in 16 (23.9%), and none in 29 (43.3%) metastases. Quantitative perfusion analysis revealed significantly higher arterial liver perfusion and HPI in metastases and metastasis borders than in adjacent normal liver tissue (P < 0.001 each). Portal-venous perfusion was significantly lower in metastases and metastasis borders than in normal liver tissue (P < 0.001). There were no significant differences in image quality and qualitative perfusion analysis between the 3 protocols (P = n.s.). Calculated effective radiation doses were 13.4 mSv for protocol A, 30.7 mSv for protocol B, and 23.0 mSv for protocol C. CT perfusion imaging of the liver using the 4D spiral-mode is feasible with diagnostic image quality, and enables the reliable qualitative and quantitative analysis of the normal and metastatic liver parenchyma. Radiation dose issues must be considered when determining the scan range, number of cycles, and scan duration of the perfusion CT protocol.


Ethylene-regulated gene expression: molecular cloning of the genes encoding an endochitinase from Phaseolus vulgaris.
A full-length copy of bean leaf chitinase mRNA has been cloned. The 1146-base-pair insert of pCH18 encodes the 27-residue amino-terminal signal peptide of the precursor and 301 residues of the mature protein. Utilizing pCH18 as a hybridization probe, we have shown that the increase in translatable chitinase mRNA seen upon ethylene treatment of bean seedlings is due to a 75- to 100-fold increase in steady-state mRNA levels. Southern blot analysis of bean genomic DNA revealed that chitinase is encoded by a small, multigene family consisting of approximately four members. From our nucleotide sequence analysis of five additional chitinase cDNA clones, it appears that at least two of these genes are expressed. Three of the bean chitinase genes have been isolated from a Sau3A genomic library and partially characterized.


The European school of total hip arthroplasty and 35 years of total hip arthroplasty in the Czech Republic.
The development of the European total hip arthroplasty in the sixties of 20th century is associated mainly with the names of G. K. McKee, J. Charnley and M. E. Muller. The SICOT meeting in Paris in 1966 introduced THA as a new method of treatment of osteoarthritis of the hip and paved the way for the commercialization of the manufacturing of prostheses. The success of this method consisted primarily in the fixation of individual components by bone cement and application of metal-to-polyethylene and metal-to-metal joints. The book presents these historical events as they have been reflected in the development of THA in the countries of the former Czechoslovakia and in the development of implants of domestic provenance. The development of THA in the former Czechoslovakia started in 1969 and was based on the Swiss school established by M. E. Muller. The Muller component with the "banana-shaped" stem was implanted using the Watson-Jones approach, the proximal femur canal was prepared by a rasp and the Palacos cement was used. This prosthesis served also as the model for the first Czech implant Poldi-Cech of I generation (the Chirulen cup available in three sizes, the "banana- shaped" femoral component with a 32 mm head-diameter and 130 degree neck-shaft angle) the development of which started in 1969 and the serial production began in 1972 in the Poldi Kladno steel company. Fatigue fractures of the "banana-shaped" stem led to the development of the concept of a femoral component with an "anatomical" stem. In Switzerland, this concept was developed by B. G. Weber. In the former Czechoslovakia the development of a new femoral component with an "anatomical" stem following the shape of the intramedullary canal with an ovoid profile and without sharp edges started in 1972. Due to the incidence of fatigue fractures of the "banana-shaped"stem and material that was not sufficiently strong (steel used for the production of osteosynthetic components), 144 degree neck-shaft angle was chosen. Reduction of the effect of bending forces on the valgus stem had eliminated fatigue fractures. This Poldi-Cech prosthesis of II generation was provided in nine sizes. The diameter of the head remained the same. Three Chirulen cups were supplemented with a "flat" cup for implantation in a dysplastic acetabulum. The prosthesis was implanted with an exact instrument set (reamer for acetabular preparation, rasp for proximal femoral preparation, alignment device for accurate positioning of both the cup and the femoral component) with the use of the Palacos cement. The production of the Poldi-Cech prosthesis of II. generation started in 1974 with the fabrication of a monoblock. Since 1986 it has been developed into a modular system (14/16 cone) with the possibility to use a ceramic head of 32 mm diameter (only in the nineties the cone size was changed to 12/14 and the heads were provided also in the 28 mm diameter). The Poldi-Cech prosthesis of II generation with the "anatomical" stem has been implanted since 1974 (i. e. during 30 years) in more than 140 000 patients in the Czech and Slovak Republics. In the same year cemented monoblock hemiarthroplasty was developed on the same basis as THA using the same "anatomical" stem and 36 to 60mm head diameter. Until now more than 35,000 of these hemiarthroplasty systems have been implanted. Until 1992 the Poldi-Cech implant was practically the only available cemented total hip replacement in the former Czechoslovakia and retrospectively it may be considered a highly successful implant. In the authors' view, the further development of THA will lead in the following years to the use of implants proved by a long-term follow-up. Preference will be given to prostheses the implantation of which will require a minimal loss of the bone stock during primary surgery and which will allow a technically easy reimplantation. The continental Europe has been recently preferring hybrid prostheses while Scandinavia witnesses an evident increase in the number of implanted cemented prostheses to the detriment of the cementless and hybrid ones. Naturally, cementless prostheses will keep dominating in young patients.


Solution structure of an oligodeoxynucleotide containing the malondialdehyde deoxyguanosine adduct N2-(3-oxo-1-propenyl)-dG (ring-opened M1G) positioned in a (CpG)3 frameshift hotspot of the Salmonella typhimurium hisD3052 gene.
The refined solution structure for the ring-opened N2-(3-oxo-1-propenyl)-dG derivative of the malondialdehyde deoxyguanosine adduct M(1)G [3-(2'-deoxy-beta-D-erythro-pentofuranosyl)pyrimido[1, 2-a]purin-10(3H)-one] in d(ATCGCXCGGCATG) x d(CATGCCGCGCGAT) [X being N(2)-(3-oxo-1-propenyl)-dG], containing the d(CpG)(3) frameshift hotspot of the Salmonella typhimurium hisD3052 gene, is presented. When inserted into this duplex, M(1)G underwent spontaneous ring opening to N2-(3-oxo-1-propenyl)-dG. NMR analysis revealed that N2-(3-oxo-1-propenyl)-dG induced minor structural perturbations in the hisD3052 oligodeoxynucleotide. However, the stability of the duplex DNA was reduced; the N2-(3-oxo-1-propenyl)-dG-modified hisD3052 oligodeoxynucleotide exhibited a 14 degrees C decrease in T(m) relative to that of the native oligodeoxynucleotide. The modified guanine maintained stacking interactions with neighboring bases but was not Watson-Crick hydrogen bonded. A total of 13 NOEs were observed from the 3-oxo-1-propenyl moiety protons of N2-(3-oxo-1-propenyl)-dG to DNA protons. Molecular dynamics calculations, restrained by 602 distance restraints derived from experimental NOE measurements and 23 empirical distance restraints, converged with pairwise rmsd differences of <0.90 A. The sixth-root residual factor with the NMR data was 9.1 x 10(-2). The cytosine complementary to N2-(3-oxo-1-propenyl)-dG was pushed toward the major groove but maintained partial stacking interactions with its neighboring bases. The modified guanine remained in the anti conformation, while the 3-oxo-1-propenyl moiety was positioned in the minor groove of the duplex. Possible correlations between the relatively small structural perturbations induced in this DNA duplex by N2-(3-oxo-1-propenyl)-dG and the mutagenic spectrum of M(1)G are discussed.


Stereochemical elucidation of the 1,4 polyketide amphidinoketide I.
The relative and absolute stereochemistry of amphidinoketide I has been determined by the total synthesis of all the diastereoisomers. Molecular modelling suggests that the natural product is not the thermodynamically preferred diastereoisomer.


Explaining racial variation in lower extremity amputation: a 5-year retrospective claims data and medical record review at an urban teaching hospital.
Population-based hospital data indicate that African American patients undergo major lower extremity (LE) amputation 2 to 3 times more frequently than white patients. Some have attributed this to a lack of access to LE revascularization procedures by African American patients. To determine the likelihood that racial disparities in amputation rates are related to treatment choice, this study examines rates of primary amputation (major amputation without any previous attempt at revascularization) and repeat amputation. Two-step case-control study, reviewing experience at a large midwestern teaching hospital. First, administrative discharge data for all 1127 patients undergoing LE arterial bypass graft, angioplasty, or major amputation from January 1, 1995, to February 1, 2000, were used to analyze racial differences in the risk of admission for major amputation vs revascularization. Medical records were then reviewed for an approximate full sample of 60 African American major amputees and a random sample of 60 (two thirds of the total) white major amputees. Racial disparities in frequency of primary and repeat amputation were analyzed, controlling for age, sex, and diabetes mellitus status. Among all patients admitted for LE ischemia, outcome measures were the odds of amputation vs revascularization, and among a sample of African American and white amputees, the odds of primary vs repeat major amputation. Among all patients hospitalized for LE ischemia, African American patients were younger (P<.05), more often female (P<.01), and more likely to undergo major amputation (odds ratio, 1.68; P =.005). However, after adjusting for age, sex, and diabetes mellitus prevalence, the analysis revealed an equal likelihood of primary amputation among African Americans and whites. Repeat amputees were 2.5 times more likely to be African American than white (P =.04). The racial disparity at the study institution was primarily due to African American patients undergoing repeat major amputation at a significantly higher rate than whites.


IFIH1 (MDA5) is required for innate immune detection of intron-containing RNA expressed from the HIV-1 provirus.
Antiretroviral therapy (ART) suppresses HIV-1 viremia and prevents progression to AIDS. Nonetheless, chronic inflammation is a common problem for people living with HIV-1 on ART. One possible cause of inflammation is ongoing transcription from HIV-1 proviruses, whether or not the sequences are competent for replication. Previous work has shown that intron-containing RNA expressed from the HIV-1 provirus in primary human blood cells, including CD4+ T cells, macrophages, and dendritic cells, activates type 1 interferon. This activation required HIV-1 rev and was blocked by the XPO1 (CRM1)-inhibitor leptomycin. To identify the innate immune receptor required for detection of intron-containing RNA expressed from the HIV-1 provirus, a loss-of-function screen was performed with shRNA-expressing lentivectors targeting twenty-one candidate genes in human monocyte derived dendritic cells. Among the candidate genes tested, only knockdown of XPO1 (CRM1), IFIH1 (MDA5), or MAVS prevented activation of the IFN-stimulated gene ISG15. The importance of IFIH1 protein was demonstrated by rescue of the knockdown with non-targetable IFIH1 coding sequence. Inhibition of HIV-1-induced ISG15 by the IFIH1-specific Nipah virus V protein, and by IFIH1-transdominant inhibitory CARD-deletion or phosphomimetic point mutations, indicates that IFIH1 filament formation, dephosphorylation, and association with MAVS, are all required for innate immune activation in response to HIV-1 transduction. Since both IFIH1 and DDX58 (RIG-I) signal via MAVS, the specificity of HIV-1 RNA detection by IFIH1 was demonstrated by the fact that DDX58 knockdown had no effect on activation. RNA-Seq showed that IFIH1-knockdown in dendritic cells globally disrupted the induction of IFN-stimulated genes. Finally, specific enrichment of unspliced HIV-1 RNA by IFIH1 was revealed by formaldehyde crosslinking immunoprecipitation (f-CLIP). These results demonstrate that IFIH1 is required for innate immune activation by intron-containing RNA from the HIV-1 provirus, and potentially contributes to chronic inflammation in people living with HIV-1.


Quantitative rather than qualitative differences in gene expression predominate in intestinal cell maturation along distinct cell lineages.
Several cell types are present in the intestinal epithelium that likely arise from a common precursor, the stem cell, and each mature cell type expresses a unique set of genes that characterizes its functional phenotype. Although the process of differentiation is intimately linked to the cessation of proliferation, the mechanisms that dictate intestinal cell fate determination are not well characterized. To investigate the reprogramming of gene expression during the cell lineage allocation/differentiation process, we took advantage of a unique system of two clonal derivatives of HT29 cells, Cl16E and Cl19A cells, which spontaneously differentiate as mucus producing goblet and chloride-secreting cells, respectively, as a function of time. By profiling gene expression, we found that these two cell lines show remarkably similar kinetics of change in gene expression and common clusters of coordinately regulated genes. This demonstrates that lineage-specific differentiation of intestinal epithelial cells is characterized overall by the sequential recruitment of functionally similar gene sets independent of the final phenotype of the mature cells.


A critical role of PRDM14 in human primordial germ cell fate revealed by inducible degrons.
PRDM14 is a crucial regulator of mouse primordial germ cells (mPGCs), epigenetic reprogramming and pluripotency, but its role in the evolutionarily divergent regulatory network of human PGCs (hPGCs) remains unclear. Besides, a previous knockdown study indicated that PRDM14 might be dispensable for human germ cell fate. Here, we decided to use inducible degrons for a more rapid and comprehensive PRDM14 depletion. We show that PRDM14 loss results in significantly reduced specification efficiency and an aberrant transcriptome of hPGC-like cells (hPGCLCs) obtained in vitro from human embryonic stem cells (hESCs). Chromatin immunoprecipitation and transcriptomic analyses suggest that PRDM14 cooperates with TFAP2C and BLIMP1 to upregulate germ cell and pluripotency genes, while repressing WNT signalling and somatic markers. Notably, PRDM14 targets are not conserved between mouse and human, emphasising the divergent molecular mechanisms of PGC specification. The effectiveness of degrons for acute protein depletion is widely applicable in various developmental contexts.


Multiomic Characterization Reveals a Distinct Molecular Landscape in Young-Onset Pancreatic Cancer.
Using a real-world database with matched genomic-transcriptomic molecular data, we sought to characterize the distinct molecular correlates underlying clinical differences between patients with young-onset pancreatic cancer (YOPC; younger than 50 years) and patients with average-onset pancreatic cancer (AOPC; 70 years and older). We analyzed matched whole-transcriptome and DNA sequencing data from 2,430 patient samples (YOPC, n = 292; AOPC, n = 2,138) from the Caris Life Sciences database (Phoenix, AZ). Immune deconvolution was performed using the quanTIseq pipeline. Overall survival (OS) data were obtained from insurance claims (n = 4,928); Kaplan-Meier estimates were calculated for age- and molecularly defined cohorts. Significance was determined as FDR-corrected P values (Q) < .05. Patients with YOPC had higher proportions of mismatch repair-deficient/microsatellite instability-high, BRCA2-mutant, and PALB2-mutant tumors compared with patients with AOPC, but fewer SMAD4-, RNF43-, CDKN2A-, and SF3B1-mutant tumors. Notably, patients with YOPC demonstrated significantly lower incidence of KRAS mutations compared with patients with AOPC (81.3% v 90.9%; Q = .004). In the KRAS wild-type subset (n = 227), YOPC tumors demonstrated fewer TP53 mutations and were more likely driven by NRG1 and MET fusions, whereas BRAF fusions were exclusively observed in patients with AOPC. Immune deconvolution revealed significant enrichment of natural killer cells, CD8+ T cells, monocytes, and M2 macrophages in patients with YOPC relative to patients with AOPC, which corresponded with lower rates of HLA-DPA1 homozygosity. There was an association with improved OS in patients with YOPC compared with patients with AOPC with KRAS wild-type tumors (median, 16.2 [YOPC-KRASWT] v 10.6 [AOPC-KRASWT] months; P = .008) but not KRAS-mutant tumors (P = .084). In this large, real-world multiomic characterization of age-stratified molecular differences in pancreatic ductal adenocarcinoma, YOPC is associated with a distinct molecular landscape that has prognostic and therapeutic implications.


Kawasaki disease: an update.
Kawasaki disease is the leading cause of acquired heart disease in children in many parts of the world. It is an acute systemic vasculitis of unknown etiology predominantly affecting children under 5 years of age. It is diagnosed clinically by the presence of a constellation of manifestations and exclusion of other diseases with similar findings. Children develop a spectrum of cardiovascular manifestations including coronary artery involvement in 30-35% of untreated cases with myocardial infarction and death in < or = 2% of them. Acute stage treatment is by administration of intravenous immunoglobulin and aspirin. Those with coronary artery involvement need long-term cardiac risk stratification and management because many of them will become the part of a growing pool of adults with ischemic heart disease.


Microbial abundance in surface ice on the Greenland Ice Sheet.
Measuring microbial abundance in glacier ice and identifying its controls is essential for a better understanding and quantification of biogeochemical processes in glacial ecosystems. However, cell enumeration of glacier ice samples is challenging due to typically low cell numbers and the presence of interfering mineral particles. We quantified for the first time the abundance of microbial cells in surface ice from geographically distinct sites on the Greenland Ice Sheet (GrIS), using three enumeration methods: epifluorescence microscopy (EFM), flow cytometry (FCM), and quantitative polymerase chain reaction (qPCR). In addition, we reviewed published data on microbial abundance in glacier ice and tested the three methods on artificial ice samples of realistic cell (10(2)-10(7) cells ml(-1)) and mineral particle (0.1-100 mg ml(-1)) concentrations, simulating a range of glacial ice types, from clean subsurface ice to surface ice to sediment-laden basal ice. We then used multivariate statistical analysis to identify factors responsible for the variation in microbial abundance on the ice sheet. EFM gave the most accurate and reproducible results of the tested methodologies, and was therefore selected as the most suitable technique for cell enumeration of ice containing dust. Cell numbers in surface ice samples, determined by EFM, ranged from ~ 2 × 10(3) to ~ 2 × 10(6) cells ml(-1) while dust concentrations ranged from 0.01 to 2 mg ml(-1). The lowest abundances were found in ice sampled from the accumulation area of the ice sheet and in samples affected by fresh snow; these samples may be considered as a reference point of the cell abundance of precipitants that are deposited on the ice sheet surface. Dust content was the most significant variable to explain the variation in the abundance data, which suggests a direct association between deposited dust particles and cells and/or by their provision of limited nutrients to microbial communities on the GrIS.


Fractional Fourier transform of Ince-Gaussian beams.
Ince-Gaussian beams are introduced to describe the natural resonating modes produced by stable resonators, and they form the third completely orthogonal family of exact solutions of the paraxial wave equation. The fractional Fourier transform (FRFT) is applied to treat the propagation of Ince-Gaussian beams, and an analytical expression for an Ince-Gaussian beam passing through a FRFT system is derived. The normalized intensity distribution of an Ince-Gaussian beam in the FRFT plane is graphically illustrated with numerical examples, and the influences of the different parameters on the normalized intensity distribution are discussed in detail.


Elevated Concentrations of Lead in Particulate Matter on the Neighborhood-Scale in Delhi, India As Determined by Single Particle Analysis.
High mass concentrations of atmospheric lead particles are frequently observed in the Delhi, India metropolitan area, although the sources of lead particles are poorly understood. In this study, particles sampled across Delhi (August - December 2008) were analyzed by computer-controlled scanning electron microscopy with energy dispersive X-ray spectroscopy (CCSEM-EDX) to improve our understanding of the spatial and physicochemical variability of lead-rich particles (>90% lead). The mean mass concentration of lead-rich particles smaller than 10 μm (PM10) was 0.7 μg/m(3) (1.5 μg/m(3) std. dev.) with high variability (range: 0-6.2 μg/m(3)). Four samples (16% of 25 samples) with PM10 lead-rich particle concentrations >1.4 μg/m(3) were defined as lead events and studied further. The temporal characteristics, heterogeneous spatial distribution, and wind patterns of events, excluded regional monsoon conditions or common anthropogenic sources from being the major causes of the lead events. Individual particle composition, size, and morphology analysis indicate informal recycling operations of used lead-acid batteries as the likely source of the lead events. This source is not typically included in emission inventories, and the observed isolated hotspots with high lead concentrations could represent an elevated exposure risk in certain neighborhoods of Delhi.


Fully intermittent dosing with drugs for tuberculosis.
The number of people infected with tuberculosis continues to rise world-wide. Rifampicin-containing treatment regimens can achieve high cure rates. Intermittent drug treatment delivered in the community has the potential to improve adherence to treatment. The objective of this review was to compare the effectiveness of rifampicin-containing short-course chemotherapy regimens, given two or three times a week, with similar regimens given daily in patients with pulmonary tuberculosis. We searched the Cochrane Infectious Diseases Group trials register, the Cochrane Controlled Trials Register, Medline, and reference lists of articles. We contacted experts in the field. Randomised and quasi-randomised trials of any multi-drug regimen containing rifampicin in patients with confirmed pulmonary tuberculosis. Treatment had to be given up to three times a week for up to nine months, with any initial daily dosing period not more than one month, and was compared to daily dosing throughout for the same period. Two reviewers independently assessed trial eligibility and quality. One trial involving 399 patients was included. The trial compared treatment three times per week with daily treatment for six months. There was no difference in cure rate (198 out of 199 people in the intermittent group compared to all 200 in the daily group), but 5 patients relapsed in the group receiving intermittent therapy compared to one in the group receiving the daily regimen. There is not enough evidence to assess the equivalence of effect between fully intermittent, rifampicin-containing short-course chemotherapy and similar daily therapy in patients with pulmonary tuberculosis. Larger randomised studies are required to establish the equivalence of fully intermittent, short-course chemotherapy, with daily regimens.


Characteristics and wounding effects of a black powder handgun.
Commercial exploitation of the recent increase in interest in American history has led to an increased availability of weapons designed to use black powder (gunpowder). In some states, controls on these weapons are poor or nonexistent. In this study a .44-caliber black powder revolver is described and compared with a .45 ACP modern revolver. The kinetic energies of their missiles were very similar. In addition to the usual wound appearances determined by range and direction of fire, it was noted that the soot associated with wounds from the black powder handgun was much greater than from the control and had a characteristic sulfurous smell. Much more true tattooing was present around the black powder wounds. The spherical shape of the missile and the presence of black powder in crypts in the missile base were both characteristic of a black powder revolver. Striations were well represented on the ball but were easily destroyed during recovery because the lead was so soft. Swabs from around the wounds and from the shooter's hand revealed a large deposit of barium but little antimony. Wounds inflicted by black powder handguns may be detected more or less reliably.


Regenerative Potential of Granulocyte Colony-Stimulating Factor Immobilized by Using Electron-Beam Synthesis Nanotechnology in an Experimental Model of Ovarian Reserve Depletion.
The pharmacological activity of granulocyte CSF (G-CSF) immobilized using electron-beam synthesis nanotechnology (imG-CSF) was evaluated in an experimental model of ovarian reserve depletion. The effectiveness of the drug was compared with that of its unmodified form. Depletion of the ovarian follicular pool in female Sprague-Dawley rats was caused by a single intravenous injection of the antitumor drug etoposide in the maximum tolerated dose. The effectiveness of the studied drugs was assessed by serum concentration of anti-Mullerian hormone (AMH) measured by ELISA and by the number of primordial, two-layer, multilayer, and atretic follicles counted on serial sections of the ovaries (5-μm thick; through the entire organ) stained with hematoxylin and eosin. It was found that imG-CSF prevents depletion of the ovarian reserve in the model used, which was confirmed by high AMH concentration and higher numbers of primordial, two- and multilayer follicles in comparison with the corresponding parameters in the control (etoposide), and by a decrease in the severity of atretic processes. Unmodified form of the drug demonstrated lower efficiency.


The Affordability of Flood Risk Property-Level Adaptation Measures.
The affordability of property-level adaptation measures against flooding is crucial due to the movement toward integrated flood risk management, which requires the individuals threatened by flooding to actively manage flooding. It is surprising to find that affordability is not often discussed, given the important roles that affordability and social justice play regarding flood risk management. This article provides a starting point for investigating the potential rate of unaffordability of flood risk property-level adaptation measures across Europe using two definitions of affordability, which are combined with two different affordability thresholds from within flood risk research. It uses concepts of investment and payment affordability, with affordability thresholds based on residual income and expenditure definitions of unaffordability. These concepts, in turn, are linked with social justice through fairness concerns, in that, all should have equal capability to act, of which affordability is one avenue. In doing so, it was found that, for a large proportion of Europe, property owners generally cannot afford to make one-time payment of the cost of protective measures. These can be made affordable with installment payment mechanisms or similar mechanisms that spread costs over time. Therefore, the movement toward greater obligations for flood-prone residents to actively adapt to flooding should be accompanied by socially accessible financing mechanisms.


The temporary postnatal decline in glucuronidation of certain phenols by rat liver.
A temporary but marked postnatal decline in UDP-glucuronosyltransferase activity occurs in homogenates and microsomes from rat liver. The profile of this trough and its time of occurrence (maximal over 13-16 days) are almost identical with the two substrates 2-aminophenol and 1-naphthol, whose rates of glucuronidation differ 10-fold. The trough is greatest with digitonin-activated preparations, least with fresh latent ('native') enzyme and intermediate when the native enzyme is treated with its specific activator UDP-N-acetylglucosamine (UDP-GlcNAc). Less detailed evidence supports similar conclusions with 4-nitrophenol as substrate. The trough is not due to the presence of an inhibitor of the transferase in rat liver at 15 days of age. Over the whole perinatal period, including the time of the trough, the enzyme in homogenates can be activated by UDP-GlcNAc; the microsomal enzyme is activated to a rather lesser degree perinatally, and evidence suggests this may be due to artefacts introduced during tissue fractionation. When the overall process of glucuronidation is studied in snips of intact liver offered high concentrations of the two different phenols, the trough is again evident over the same period as observed with broken cells, and of equal depth for both substrates. The infant rat is therefore probably less able to glucuronidate hepatically these phenols over the suckling or early weaning period than are the adult, late foetus or newborn, and may be especially incompetent at 13-16 days of age.


Penile prosthesis implant for primary erectile dysfunction in patient with Klippel-Trenaunay syndrome complicated by consumptive coagulopathy: A case report.
Klippel-Trenaunay syndrome (KTS) is a congenital disorder characterized by cutaneous port wine capillary malformations, varicose veins with hemihypertrophy of soft tissue and bone.Pelvic and retroperitoneal vascular malformations have been described up to the 30% of patients with KTS while hemangiomas of the urinary tract have been reported in 6% PATIENT CONCERNS:: A 30-year-old man with KTS was referred to our center for primary erectile dysfunction (ED) associated with varicosities of unusual distribution and asymmetry of the lower limbs. Furthermore, he suffered from hypertension and autosomal dominant polycystic kidney disease.During penile prosthesis implantation, a significant intraoperative bleeding (1 liter) due to large scrotal venous malformations and profuse bleeding from both corpora was recorded. One month later, the day after the first prosthesis training session, the patient returned with swelling in the penoscrotal region. A large inguino-scrotal hematoma was drained. There was a complete bilateral dehiscence of corpora cavernosa with a spread venous bleeding in the scrotum. CT scan showed hypertrophy of the right hypogastric artery with severe vascular malformations: the right pudendal artery was massively dilated with early visualization of venous drainage without evidence of arteriovenous fistulae; regular bulbocavernous capillary blush; right upper gluteus artery hypertrophic and dilated. Multiple twisting and aneurysms of the right internal pudenda artery were bleeding from multiple points. Cystoscopy showed a fistula between the proximal urethra and the penoscrotal dartos. Coagulation tests revealed the presence of factor XIII deficiency INTERVENTIONS:: The patient underwent several procedures including percutaneous scleroembolization of the internal pudendal arteries, removal of the penile implant, recombinant factor XIII (FXIII) administration, and cord blood platelet gel application. The patient was discharged after almost 3 months in hospital, hemodynamically stable. Experience regarding management of ED in KTS patient is limited and in case of concomitant factor XIII deficiency, the clinical scenario can be life-threatening. A multidisciplinary approach including a urologist, an interventional radiologist and a hematologist in our experience represented the key approach in case of severe bleeding following surgery for ED.


Inflorescence commitment and architecture in Arabidopsis.
Flowering plants exhibit one of two types of inflorescence architecture: indeterminate, in which the inflorescence grows indefinitely, or determinate, in which a terminal flower is produced. The indeterminate condition is thought to have evolved from the determinate many times, independently. In two mutants in distantly related species, terminal flower 1 in Arabidopsis and centroradialis in Antirrhinum, inflorescences that are normally indeterminate are converted to a determinate architecture. The Antirrhinum gene CENTRORADIALIS (CEN) and the Arabidopsis gene TERMINAL FLOWER 1 (TFL1) were shown to be homologous, which suggests that a common mechanism underlies indeterminacy in these plants. However, unlike CEN, TFL1 is also expressed during the vegetative phase, where it delays the commitment to inflorescence development and thus affects the timing of the formation of the inflorescence meristem as well as its identity.


A Preliminary Study of the Complement Component 1q Levels in Predicting the Efficacy of Combined Immunotherapy in Patients with Lung Cancer.
To evaluate the value of serum complement component 1q (C1q) levels in predicting the efficacy of combined immunotherapy in patients with lung cancer. A total of 42 patients with lung cancer who received combined immunotherapy in the cancer center of Renmin Hospital of Wuhan University were included in this study. The clinical data of serum C1q and lactate dehydrogenase (LDH) levels before and three weeks after immunotherapy were collected. Response evaluation showed that the number of patients with complete response (CR), partial response (PR), stable disease (SD), and progressive disease (PD) was 0 (0%), 26 (61.9%), 14 (33.3%), and 2 (4.8%), respectively. The CR/PR group (patients with CR or PR) showed higher pC1q (C1q level before immunotherapy) and iC1q (C1q level 3 weeks after immunotherapy) than the SD/PD group (patients with SD or PD). The LDH reduction (96.2%) and C1q increment (84.6%) in the CR/PR group 3 weeks after immunotherapy were higher than those of the SD/PD group, and the differences were statistically significant. Logistic regression analysis indicated that pC1q, iC1q, and LDH level trends 3 weeks after the treatment were significantly correlated to the efficacy of combined immunotherapy with odds ratios of 8.185, 5.500, and 0.031, respectively. High C1q levels before immunotherapy and increased C1q levels and decreased LDH levels 3 weeks afterward suggest good therapeutic effects of combined immunotherapy in patients with lung cancer. Serum C1q levels have certain clinical significance in predicting the efficacy of combined immunotherapy.


In vivo 2-deoxyglucose administration preserves glucose and glutamate transport and mitochondrial function in cortical synaptic terminals after exposure to amyloid beta-peptide and iron: evidence for a stress response.
Mild metabolic stress can increase resistance of neurons in the brain to subsequent more severe insults, as exemplified by the beneficial effects of heat shock and ischemic preconditioning. Studies of Alzheimer's disease and other age-related neurodegenerative disorders indicate that dysfunction and degeneration of synapses occur early in the cell death process, and that oxidative stress and mitochondrial dysfunction are central events in this pathological process. It was recently shown that administration of 2-deoxy-d-glucose (2DG), a nonmetabolizable glucose analog that induces metabolic stress, to rats and mice can increase resistance of neurons in the brain to excitotoxic, ischemic, and oxidative injury. We now report that administration of 2DG to adult rats (daily i.p. injections of 100 mg/kg body weight) increases resistance of synaptic terminals to dysfunction and degeneration induced by amyloid beta-peptide and ferrous iron, an oxidative insult. The magnitude of impairment of glucose and glutamate transport induced by amyloid beta-peptide and iron was significantly reduced in cortical synaptosomes from 2DG-treated rats compared to saline-treated control rats. Mitochondrial dysfunction, as indicated by increased levels of reactive oxygen species and decreased transmembrane potential, was significantly attenuated after exposure to amyloid beta-peptide and iron in synaptosomes from 2DG-treated rats. Levels of the stress proteins HSP-70 and GRP-78 were increased in synaptosomes from 2DG-treated rats, suggesting a mechanism whereby 2DG protects synaptic terminals. We conclude that 2DG bolsters cytoprotective mechanisms within synaptic terminals, suggesting novel preventative and therapeutic approaches for neurodegenerative disorders.


Prevalence and risk factors of hepatocellular carcinoma in Japanese patients with primary biliary cirrhosis.
Primary biliary cirrhosis (PBC) tends to affect females more than males. PBC selectively damages intrahepatic small bile ducts, particularly interlobular bile ducts. The clinical presentation of PBC has changed according to recent advances in clinicobiological diagnosis and improvements in therapeutic effects and prognosis. In particular, we encounter PBC patients with hepatocellular carcinoma (HCC), and the number of these patients appears to have increased. The precise reason for the increased number of PBC patients with HCC in recent decades remains unknown, but recognizing the current status of carcinogenesis in PBC patients, identifying the associated clinicopathological risk factors and understanding how the pathogenesis of PBC is directly associated with HCC, is important. In this review, we summarize the data from two nationwide surveys undertaken in Japan as well as recent data from Japanese and international studies.


PCB concentrations and dioxin-like activity in blood samples from Danish school children and their mothers living in urban and rural areas.
Human exposure to persistent organic pollutants (POPs) is of major concern due to a diversity of adverse effects from prolonged exposure and bioaccumulation. Manufacturing of polychlorinated biphenyls (PCBs), a subgroup of POPs, has been prohibited for many decades; however, human exposure still occurs due to the persistent nature of the chemicals. The concentrations of the dioxin-like PCB congeners 105, 118 and 156 and the non-dioxin-like PCB congeners 28, 52, 101, 138, 153 and 180, p,p'-DDE, p,p'-DDT, o,p'-DDE, o,p'-DDT, HCB and β-HCH as well as the dioxin-like activity using the AhR transactivity assay were analysed in blood samples from Danish schoolchildren and their mothers in the European framework of the DEMOCOPHES/COPHES projects. The participants were selected from an urban and a rural area, respectively. The PCB concentrations and the AhR-TEQ (TCDD toxic equivalent) were significantly higher in schoolchildren living in the urban area compared with the rural, and for AhR-TEQ, a strong correlation between the mothers and children was observed. We found a significant negative correlation between BMI and PCB concentrations in the children. Finally, in the mothers, there was a positive association between age and PCB concentration. These results show that both PCBs and dioxin-like activity can be measured as biomarkers of exposure and effects in blood samples from children and women. The results indicate that people living in urban areas may be exposed to higher concentrations of PCBs, dioxins and dioxin-like chemicals, which may lead to a greater risk of adverse effects for urban populations.


Clinical outcome of intentional replantation with preoperative orthodontic extrusion: a retrospective study.
To evaluate retrospectively the clinical outcomes of intentional replantation (IR) of teeth in terms of tooth survival and periradicular healing and to investigate their prognostic factors. Two hundred and eighty-seven teeth treated by IR were analysed retrospectively. Clinical outcomes between the cases with preoperative orthodontic extrusion for 2-3 week and those without extrusion were analysed. The outcomes of IR were determined by clinical and radiographic evaluation. Tooth survival and periradicular healing estimates were compared using Kaplan-Meier analysis. The contribution of a patient's age and gender, tooth type and location, and preoperative orthodontic extrusion was investigated using the multivariate Cox proportional hazard model. The mean follow-up period was 25.4 ± 9.3 months. The overall success rate of IR was 89.5% based on periradicular healing. The overall survival rate was 95.1%. The survival rates were 91.2% for the teeth extracted without extrusion and 98.1% for those extracted with extrusion. Amongst the variables tested, only the extraction technique with preoperative orthodontic extrusion significantly affected the survival rate (P = 0.016). Other prognostic variables, such as age and gender, tooth type and location, did not affect the survival of intentionally replanted teeth. Intentional replantation was a viable treatment option for teeth with previously failed nonsurgical root canal treatment, regardless of a patient's age and gender, and tooth type and location. Preoperative orthodontic extrusion for 2-3 weeks reduced root resorption and tooth fracture and can be recommended to enhance the outcome of IR.


Isolation of novel, transcriptionally active AP-1 binding sites: implications for cellular transformation.
Increased AP-1 DNA-binding activity, in the context of TRE-binding, is not a consequence of Fos transformation. In this report we investigate the possibility of a change in binding site preference by vFosAP-1 compared with AP-1 from an untransformed cell. Fos binding sites were immunoselected from random sequence oligonucleotides using a pan Fos anti-serum with nuclear protein from quiescent FBRp75v-fos-transformed (FBR) and normal (208F) rat fibroblasts. The selected oligonucleotides were aligned by computer and a consensus described for the sequences bound by AP-1 from the two cell lines. The vFos binding site is shown to be a consensus TRE, whereas the sequence ACCACATC is described as the cellular Fos protein family consensus. We demonstrate that sequences differing from the TRE consensus can bind AP-1 and direct transcription. AP-1 DNA-binding activity differs between normal and transformed cells with several of the selected oligonucleotides. These sequences also demonstrate differential transcriptional activation between normal and transformed cells. In particular, the 208F consensus has no transcriptional activity in FBR cells. Further, EGF differentially influences the transcriptional activity of the oligonucleotides in 208F and FBR cells. Our results suggest that AP-1 may change its preferred binding site depending on the proteins available at any given time, the sequences flanking a non-consensus TRE or even the environment in which the cell exists. These differences in binding site preference and transcriptional activation may result in the increased transforming ability of the v-fos oncogene compared with the c-fos proto-oncogene and may extend the potential target genes beyond those with an AP-1 consensus binding site.


Using the life history model to set the stage(s) of growth and senescence in bioarchaeology and paleodemography.
Paleodemography, the study of demographic parameters of past human populations, relies on assumptions including biological uniformitarianism, stationary populations, and the ability to determine point age estimates from skeletal material. These assumptions have been widely criticized in the literature and various solutions have been proposed. The majority of these solutions rely on statistical modeling, and have not seen widespread application. Most bioarchaeologists recognize that our ability to assess chronological age is inherently limited, and have instead resorted to large, qualitative, age categories. However, there has been little attempt in the literature to systematize and define the stages of development and ageing used in bioarchaeology. We propose that stages should be based in the human life history pattern, and their skeletal markers should have easily defined and clear endpoints. In addition to a standard five-stage developmental model based on the human life history pattern, current among human biologists, we suggest divisions within the adult stage that recognize the specific nature of skeletal samples. We therefore propose the following eight stages recognizable in human skeletal development and senescence: infancy, early childhood, late childhood, adolescence, young adulthood, full adulthood, mature adulthood, and senile adulthood. Striving toward a better prediction of chronological ages will remain important and could eventually help us understand to what extent past societies differed in the timing of these life stages. Furthermore, paleodemographers should try to develop methods that rely on the type of age information accessible from the skeletal material, which uses life stages, rather than point age estimates.


Negative feedback regulation of Met-dependent invasive growth by Notch.
The hepatocyte growth factor (HGF) receptor encoded by the Met oncogene controls a genetic program-known as "invasive growth"-responsible for several developmental processes and involved in cancer invasion and metastasis. This program functions through several regulatory gene products, as yet largely unknown, both upstream and downstream of Met. Here we show that activation of the Notch receptor results in transcriptional down-regulation of Met, suppression of HGF-dependent Ras signaling, and impairment of HGF-dependent cellular responses. In turn, Met activation leads to transcriptional induction of the Notch ligand Delta and the Notch effector HES-1, indicating that Met is able to self-tune its own protein levels and the ensuing biochemical and biological outputs through stimulation of the Notch pathway. By using branching morphogenesis of the tracheal system in Drosophila as a readout of invasive growth, we also show that exogenous expression of a constitutively active form of human Met induces enhanced sprouting of the tracheal tree, a phenotype that is further increased in embryos lacking Notch function. These results unravel an in-built mechanism of negative feedback regulation in which Met activation leads to transcriptional induction of Notch function, which in turn limits HGF activity through repression of the Met oncogene.


Hepatocellular carcinoma: an evolving challenge in viral hepatitis and HIV coinfection.
Liver-related morbidity and mortality is expanding in people living with HIV. Hepatocellular carcinoma (HCC), the third most lethal malignancy on a global scale, is a dominant complication of chronic liver disease and cirrhosis in patients with coexisting hepatitis. HIV infection further complicates the clinical heterogeneity of HCC, posing concurrent challenges stemming from the underlying immunological status of the patients and the ongoing need for combined antiretroviral therapy. In this article, we review the multiple clinical implications that characterize the multidisciplinary management of HCC in the context of HIV infection.


Long-term growth hormone treatment in growth hormone deficient adults.
Growth hormone treatment in GH-deficient adults has proved beneficial in recent short-term trials, but long-term results have not yet been reported. Thirteen GH-deficient adults (4 females, 9 males; mean (SEM) age 26.4 (1.7) years), who had completed 4 months of GH therapy in a double-blind placebo-controlled cross-over study were followed, for further 16.1 (0.8) months of uninterrupted GH therapy in an open design. A significant mean increase of 1.3 cm in linear height was recorded, whereas body mass index remained unchanged. Mean muscle volume of the thigh, estimated by computerised tomography, increased significantly compared with that of the initial placebo period (p = 0.01), and a slight decrease was recorded in adipose tissue volume of the thigh (p = 0.10) and subscapular skinfold thickness (p = 0.10). Still, the muscle to fat ratio of the thigh was significantly lower compared with that of normal subjects (72.6/27.4 vs 77.9/22.1) (p less than 0.01). The mean isometric strength of the quadriceps muscles increased significantly during long-term GH therapy (p less than 0.01), but remained lower compared with that of normal subjects (1.66 (0.10) vs 2.13 (0.11) Nm/kg body weight). Exercise capacity performed on a bicycle ergometer increased significantly after long-term therapy (p less than 0.05), but still did not reach the values seen in normal subjects (22.5 (3.4) vs 37.4 (4.2) watt.min.kg-1. No adverse reactions were recorded during long-term therapy and hemoglobin A1c remained unchanged. These data suggest that long-term GH replacement therapy in GH-deficient adults has beneficial effects on several physiological features which are subnormal in these patients.


Effects of Scaffolds on Urine- and Urothelial Carcinoma Tissue-Derived Organoids from Bladder Cancer Patients.
Organoids are three-dimensional constructs generated by placing cells in scaffolds to facilitate the growth of cultures with cell-cell and cell-matrix interactions close to the in vivo situation. Organoids may contain different types of cells, including cancer cells, progenitor cells, or differentiated cells. As distinct culture conditions have significant effects on cell metabolism, we explored the expansion of cells and expression of marker genes in bladder cancer cells expanded in two different common scaffolds. The cells were seeded in basement membrane extract (BME; s.c., Matrigel®) or in a cellulose-derived hydrogel (GrowDex®, GD) and cultured. The size of organoids and expression of marker genes were studied. We discovered that BME facilitated the growth of significantly larger organoids of cancer cell line RT112 (p < 0.05), cells from a solid tumor (p < 0.001), and a voiding urine sample (p < 0.001). Expression of proliferation marker Ki76, transcription factor TP63, cytokeratin CK20, and cell surface marker CD24 clearly differed in these different tumor cells upon expansion in BME when compared to cells in GD. We conclude that the choice of scaffold utilized for the generation of organoids has an impact not only on cell growth and organoid size but also on protein expression. The disadvantages of batch-to-batch-variations of BME must be balanced with the phenotypic bias observed with GD scaffolds when standardizing organoid cultures for clinical diagnoses.


[How to identify an aspecific association? Epidemiological considerations on the study of health of office workers].
An epidemiological approach to the study of health conditions of office workers. Nowadays the work-relatedness of the majority of diseases occurring at the workplace is by no means obvious. The relationship to work environment and performance of work is one of several causative factors. However, the multifactorial origin of diseases is not at all a new concept since principles and methods for the study of the numerous factors playing a role in any association between exposure and disease are already available. The model of causal constellations developed by Rothman can be particularly useful for the investigation of "non-specific" associations, like those commonly occurring in office work. The study of the health conditions of office workers is faced with many difficulties, including the identification, measurement and classification of exposure variables and health outcomes. In designing these studies, particular attention should be devoted to aspects regarding sample size, length of observation and referent population.


Autologous pericardium annuloplasty: a "physiological" mitral valve repair.
Autologous pericardium annuloplasty (APA) is an alternative to prosthetic ring implantation for mitral valve (MV) repair, avoiding the use of foreign material and preserving the mitral annulus' physiological motion. However, data on durability are questionable. Therefore, we analyzed long-term outcomes of treating degenerative mitral regurgitation (MR) with APA. Four hundred ninety patients (mean age, 54.3±11.3 years, [15-77 years]; N.=360 men [74.1%]) who had undergone APA and neochordae implantation between July 1988 and December 2006 were retrospectively studied. MR was purely degenerative in 434 (89.3%) patients; endocarditis was present in 44 (9.1%) patients; an anterior, posterior, or bileaflet prolapse was present in 32 (6.6%), 241 (49.6%), and 213 (43.8%) patients, respectively. Clinical follow-up was 100% complete at a median of 6.5 years (5th percentile, 0.9; 95th percentile, 14.9) with an echocardiographic study in 92% of patients. In-hospital mortality was 1% (5 deaths); overall and late cardiac mortality were 7.6% and 3.9% (37 and 19 deaths), respectively. Kaplan-Meier curves for overall survival, late cardiac survival, and freedom from reoperation at 15 years (20 cases) were 86% (95%CI 80-91), 93% (95%CI 88-96), and 93% (95%CI 88-96), respectively. At 15 years, freedom from recurrent MR (28 patients) and endocarditis (6 events) were 86% (95%CI 76-91) and 97% (95%CI 92-99). Dehiscence, significant calcification of APA, and hemolysis never occurred. At reoperations, annular pericardium appeared covered by a smooth layer of tissue. APA is feasible, safe, and cost-effective, providing long-term durability, high survival, and a low rate of valve-related complications.


[Histological and immunohistochemical study on 8 cases of medullary carcinoma of the thyroid].
Histological review and immunohistochemical studies of 8 cases of medullary carcinoma were carried out by using ABC technique. The results showed 8 calcitonin positive cases, 3 Somatostatin positive cases, 7 NSE positive cases, 5 CEA positive cases and 8 keratin positive cases. In addition, histogenesis, histological characteristics and the evaluation of immunohistochemistry in diagnosis of thyroid medullary carcinoma are discussed.


Pharmacologic Stepwise Multimodal Approach for Postpartum Pain Management: ACOG Clinical Consensus No. 1.
Pain in the postpartum period is common and considered by many individuals to be both problematic and persistent (1). Pain can interfere with individuals' ability to care for themselves and their infants, and untreated pain is associated with risk of greater opioid use, postpartum depression, and development of persistent pain (2). Clinicians should therefore be skilled in individualized management of postpartum pain. Though no formal time-based definition of postpartum pain exists, the recommendations presented here provide a framework for management of acute perineal, uterine, and incisional pain. This Clinical Consensus document was developed using an a priori protocol in conjunction with the authors listed. This document has been revised to incorporate more recent evidence regarding postpartum pain.


Clinical nurse specialists in a Web-based practice environment.
The clinical nurse specialists working with the Canadian Virtual Hospice (CVH) are web-based, advanced practice palliative care nursing consultants. They deal with the complexities and challenges of online communication, particularly as it relates to the sharing of information about palliative and end-of-life care with lay people and health-care professionals alike. These nurses possess a number of highly developed skills--in communication, use of evidence-based research, teaching and understanding health-care systems--that enable them to respond to questions posed by visitors to the "Ask A Professional" section of the CVH website. Although the website is administratively based in Winnipeg, Manitoba, the services of the CNSs are available to people across Canada. In this article, the authors describe how the elements of advanced practice operate within this unique delivery model.


Dx for a careful approach to moving dual-eligible beneficiaries into managed care plans.
Policy makers are moving rapidly to develop and test reforms aimed at doing a better job of managing the costs and care for people dually eligible for Medicare and Medicaid. This commentary underscores the importance of pursuing new initiatives to address care coordination and spending concerns. It then focuses on key issues raised by proposals that would shift dual-eligible beneficiaries into managed care plans. The paper describes the heterogeneity and complexity of this population, emphasizing the need for approaches closely tied to the needs of particular subgroups of dual-eligible beneficiaries. It warns against moving too quickly, noting the time and resources required to build capacity to serve patients, secure provider networks, and develop an infrastructure for integrating and managing both Medicare and Medicaid services. The commentary cautions that optimistic savings assumptions might not materialize, raises questions about how savings will be allocated, and highlights the need for accountability as new models are being developed and tested to improve care for a population with complex needs.


Clinical use of mechanical detachable coils for dural arteriovenous fistula.
Seven dural arteriovenous fistulas were successfully embolized with mechanical detachable coils. Two lesions were located in the transverse-sigmoid sinus, four in the cavernous sinus, and one in the marginal sinus. All lesions were completely occluded on postembolization angiography. No recurrent symptoms appeared during a mean follow-up period of 11 months. Owing to the length and retrievability of the mechanical detachable coils, embolization was quicker and safer, and coils were packed more densely, than is possible with conventional coils. Coil migration was avoided because coils of the appropriate size were chosen before they were placed.


Infrared metamaterial phase holograms.
As a result of advances in nanotechnology and the burgeoning capabilities for fabricating materials with controlled nanoscale geometries, the traditional notion of what constitutes an optical device continues to evolve. The fusion of maturing low-cost lithographic techniques with newer optical design strategies has enabled the introduction of artificially structured metamaterials in place of conventional materials for improving optical components as well as realizing new optical functionality. Here we demonstrate multilayer, lithographically patterned, subwavelength, metal elements, whose distribution forms a computer-generated phase hologram in the infrared region (10.6 μm). Metal inclusions exhibit extremely large scattering and can be implemented in metamaterials that exhibit a wide range of effective medium response, including anomalously large or negative refractive index; optical magnetism; and controlled anisotropy. This large palette of metamaterial responses can be leveraged to achieve greater control over the propagation of light, leading to more compact, efficient and versatile optical components.


Effect of pramlintide on prandial glycemic excursions during closed-loop control in adolescents and young adults with type 1 diabetes.
Even under closed-loop (CL) conditions, meal-related blood glucose (BG) excursions frequently exceed target levels as a result of delays in absorption of insulin from the subcutaneous site of infusion. We hypothesized that delaying gastric emptying with preprandial injections of pramlintide would improve postprandial glycemia by allowing a better match between carbohydrate and insulin absorptions. Eight subjects (4 female; age, 15-28 years; A1C, 7.5 ± 0.7%) were studied for 48 h on a CL insulin-delivery system with a proportional integral derivative algorithm with insulin feedback: 24 h on CL control alone (CL) and 24 h on CL control plus 30-μg premeal injections of pramlintide (CLP). Target glucose was set at 120 mg/dL; timing and contents of meals were identical on both study days. No premeal manual boluses were given. Differences in reference BG excursions, defined as the incremental glucose rise from premeal to peak, were compared between conditions for each meal. CLP was associated with overall delayed time to peak BG (2.5 ± 0.9 vs. 1.5 ± 0.5 h; P < 0.0001) and reduced magnitude of glycemic excursion (88 ± 42 vs. 113 ± 32 mg/dL; P = 0.006) compared with CL alone. Pramlintide effects on glycemic excursions were particularly evident at lunch and dinner, in association with higher premeal insulin concentrations at those mealtimes. Pramlintide delayed the time to peak postprandial BG and reduced the magnitude of prandial BG excursions. Beneficial effects of pramlintide on CL may in part be related to higher premeal insulin levels at lunch and dinner compared with breakfast.


Effect of chronic nifedipine therapy on the haemodynamic response to phenylephrine before and during halothane anaesthesia.
We evaluated the effect of chronic oral nifedipine therapy (30-80 mg day-1) on the haemodynamic response to the alpha 1-agonist phenylephrine (PE) in 10 patients before and during halothane anaesthesia. The response was compared to a control group of eight patients not taking any cardiovascular drugs. No patients had ganglionic, beta-adrenergic, or muscarinic pharmacological blockade as in prior studies in the literature, and thus had intact cardiovascular reflexes. In all patients, PE caused a dose-related increase in arterial pressure and fall in heart rate and cardiac index. The response in patients taking nifedipine was not significantly different from that measured in controls. One-per-cent halothane in oxygen did not alter the overall haemodynamic response to PE in either group.


Impact of computerized prescriber order entry on the incidence of adverse drug events in pediatric inpatients.
This study was conducted to determine the impact of a computerized physician order entry system with substantial decision support on the incidence and types of adverse drug events in hospitalized children. A prospective methodology was used for the collection of adverse drug events and potential adverse drug events from all patients admitted to the pediatric intensive care and general pediatric units over a 6-month period. Data from a previous adverse drug event study of the same patient care units before computerized physician order entry implementation were used for comparison purposes. Data for 1197 admissions before the introduction of computerized physician order entry were compared with 1210 admissions collected after computerized physician order entry implementation. After computerized physician order entry implementation, it was observed that the number of preventable adverse drug events (46 vs 26) and potential adverse drug events (94 vs 35) was reduced. Reductions in overall errors, dispensing errors, and drug-choice errors were associated with computerized physician order entry. There were reductions in significant events, as well as those events rated as serious or life threatening, after the implementation of computerized physician order entry. Some types of adverse drug events continued to persist, specifically underdosing of analgesics. There were no differences in length of stay or patient disposition between preventable adverse drug events and potential adverse drug events in either study period. This study demonstrated that a computerized physician order entry system with substantive decision support was associated with a reduction in both adverse drug events and potential adverse drug events in the inpatient pediatric population. Additional system refinements will be necessary to affect remaining adverse drug events. Preventable events did not predict excess length of stay and instead may represent a sign, rather than a cause, of more complicated illness.


Squamous cell carcinoma of the scrotum: long-term followup of 14 patients.
Squamous cell carcinoma of the scrotum was diagnosed in 14 patients from 1945 to 1990. Patient age at diagnosis ranged from 40 to 73 years, with the mean age of 62 years. The most common presentation was a solitary skin lesion but inguinal adenopathy was noted in 5 patients (36%). The mean delay to diagnosis for all patients was 22 months, with a range of 2 months to 10 years. Predisposing factors included psoriasis treated with coal tar or arsenic, human papillomavirus infection and multiple cutaneous epitheliomas. The primary lesion was treated by local or wide local excision in all 14 patients. In addition, 4 patients underwent inguinal lymphadenectomy and 3 underwent radiotherapy to the pelvic and inguinal lymph nodes. Mean followup for all patients was 6 years. However, 11 patients were disease-free with a mean followup of 7 years. Improved prognosis was noted in patients with locally confined disease or carcinoma in situ only. There was no correlation between grade of tumor and survival. All patients with stages A1 and B disease treated with wide local excision and/or inguinal lymphadenectomy have done well on followup. Radiotherapy does not appear to impact on survival for patients with high stage disease.


Anoctamin 6 differs from VRAC and VSOAC but is involved in apoptosis and supports volume regulation in the presence of Ca2+.
Anoctamin 6 (ANO6), also known as TMEM16F, has been shown to be a calcium-activated anion channel with delayed calcium activation. The cellular function of ANO6 is under debate, and different groups have come to different conclusions about ANO6's physiological role. Although it is now quite well established that ANO6 is distinct from the volume-regulated anion channel, it is still unclear whether ANO6 or other anoctamins can be activated by cell swelling. In this study, we suggest that ANO1, ANO6, and ANO10 do not contribute to the volume-activated current in ANO-overexpressing HEK293 cells. Furthermore, knock-down of ANO6 in Ehrlich ascites tumor cells (EATC) and Ehrlich-Lettre ascites (ELA) did not decrease but instead significantly increased swelling-activated membrane currents. Knock-down of ANO6 in EATC did not reduce regulatory volume decrease (RVD) in the absence of extracellular calcium, whereas it significantly reduced RVD in the presence of calcium. Interestingly, we found that knock-down of ANO6 in ELA cells resulted in a decrease in cisplatin-induced caspase-3 activity, confirming earlier findings that ANO6 is involved in apoptosis. Finally, knock-down of ANO1 and ANO6 did not affect the volume-sensitive release of taurine in ELA cells. Thus, our data provide evidence that ANO6 cannot be activated directly by cell swelling unless Ca(2+) is present. We also conclude that ANO6 carries a current during RVD, provided extracellular calcium is present. Thus, swelling activation of ANO6 requires the presence of free calcium.


Phylogenetic and Biochemical Analyses of Mycobacterial l,d-Transpeptidases Reveal a Distinct Enzyme Class That Is Preferentially Acylated by Meropenem.
The genomes of diverse mycobacterial species encode multiple proteins with the canonical l,d-transpeptidase (Ldt) sequence motif. The reason for this apparent redundancy is not well understood, but evidence suggests paralogous Ldts may serve niche roles in maintaining and/or remodeling mycobacterial peptidoglycan. We examined 323 mycobacterial Ldts and determined these enzymes cluster into six clades. We identified a variably represented yet distinct Ldt class (class 6) containing Mycobacterium smegmatis (Msm) LdtF and built a homology model of Msm LdtF toward elucidating class 6 structural and functional differences. We report class 6 Ldts have structurally divergent catalytic domains containing a 10-residue insertion near the active site and additionally determined that meropenem preferentially acylates LdtF. Our data demonstrate an evolutionary basis for mycobacterial Ldt multiplicity that lends support to the idea that paralogous Ldts serve nonredundant roles in vivo and suggests class 6 Ldts can be selectively targeted by specific carbapenem antibiotics.


New dopaminergic and indoleamine-accumulating cells in the growth zone of goldfish retinas after neurotoxic destruction.
Juvenile goldfish were allowed to grow for 3 months after dopaminergic or indoleamine-accumulating cells in their retinas had been destroyed by intravitreal injection of 6-hydroxydopamine or 5,7-dihydroxytryptamine, respectively. New cells of each type were found growing in concentric rings at the margin of the retina. To compensate for the loss of dopaminergic innervation in retinas treated with 6-hydroxydopamine, cells in the growth zone appeared to proliferate at a higher rate than those in untreated retinas and long processes were extended into the retina by the first dopaminergic cells to appear.


A Review of Functional Encryption in IoT Applications.
The Internet of Things (IoT) represents a growing aspect of how entities, including humans and organizations, are likely to connect with others in their public and private interactions. The exponential rise in the number of IoT devices, resulting from ever-growing IoT applications, also gives rise to new opportunities for exploiting potential security vulnerabilities. In contrast to conventional cryptosystems, frameworks that incorporate fine-grained access control offer better opportunities for protecting valuable assets, especially when the connectivity level is dense. Functional encryption is an exciting new paradigm of public-key encryption that supports fine-grained access control, generalizing a range of existing fine-grained access control mechanisms. This survey reviews the recent applications of functional encryption and the major cryptographic primitives that it covers, identifying areas where the adoption of these primitives has had the greatest impact. We first provide an overview of different application areas where these access control schemes have been applied. Then, an in-depth survey of how the schemes are used in a multitude of applications related to IoT is given, rendering a potential vision of security and integrity that this growing field promises. Towards the end, we identify some research trends and state the open challenges that current developments face for a secure IoT realization.


A framework for crafting clinical practice guidelines that are relevant to the care and management of people with multimorbidity.
Many patients of all ages have multiple conditions, yet clinicians often lack explicit guidance on how to approach clinical decision-making for such people. Most recommendations from clinical practice guidelines (CPGs) focus on the management of single diseases, and may be harmful or impractical for patients with multimorbidity. A major barrier to the development of guidance for people with multimorbidity stems from the fact that the evidence underlying CPGs derives from studies predominantly focused on the management of a single disease. In this paper, the investigators from the Improving Guidelines for Multimorbid Patients Study Group present consensus-based recommendations for guideline developers to make guidelines more useful for the care of people with multimorbidity. In an iterative process informed by review of key literature and experience, we drafted a list of issues and possible approaches for addressing important coexisting conditions in each step of the guideline development process, with a focus on considering relevant interactions between the conditions, their treatments and their outcomes. The recommended approaches address consideration of coexisting conditions at all major steps in CPG development, from nominating and scoping the topic, commissioning the work group, refining key questions, ranking importance of outcomes, conducting systematic reviews, assessing quality of evidence and applicability, summarizing benefits and harms, to formulating recommendations and grading their strength. The list of issues and recommendations was reviewed and refined iteratively by stakeholders. This framework acknowledges the challenges faced by CPG developers who must make complex judgments in the absence of high-quality or direct evidence. These recommendations require validation through implementation, evaluation and refinement.


Conservation of nodulation genes between Rhizobium meliloti and a slow-growing Rhizobium strain that nodulates a nonlegume host.
Parasponia, a woody member of the elm family, is the only nonlegume genus whose members are known to form an effective nitrogen-fixing symbiosis with a Rhizobium species. The bacterial strain RP501 is a slow-growing strain of Rhizobium isolated from Parasponia nodules. Strain RP501 also nodulates the legumes siratro (Macroptilium atropurpureum) and cowpea (Vigna unguiculata). Using a cosmid clone bank of RP501 DNA, we isolated a 13.4-kilobase (kb) EcoRI fragment that complemented insertion and point mutations in three contiguous nodulation genes (nodABC) of Rhizobium meliloti, the endosymbiont of alfalfa (Medicago sativa). The complemented R. meliloti nod mutants induced effective nitrogen-fixing nodules on alfalfa seedlings but not on siratro, cowpeas, or Parasponia. The cloned RP501 nodulation locus hybridized to DNA fragments carrying the R. meliloti nodABC genes. A 3-kb cluster of Tn5 insertion mutations on the RP501 13.4-kb EcoRI fragment prevented complementation of R. meliloti nodABC mutations.


Is single reading with computer-aided detection (CAD) as good as double reading in mammography screening? A systematic review.
In accordance with European guidelines, mammography screening comprises independent readings by two breast radiologists (double reading). CAD (computer-aided detection) has been suggested to complement or replace one of the two readers (single reading + CAD).The aim of this systematic review is to address the following question: Is the reading of mammographic x-ray images by a single breast radiologist together with CAD at least as accurate as double reading? The electronic literature search included the databases Pub Med, EMBASE and The Cochrane Library. Two independent reviewers assessed abstracts and full-text articles. 1049 abstracts were identified, of which 996 were excluded with reference to inclusion and exclusion criteria; 53 full-text articles were assessed for eligibility. Finally, four articles were included in the qualitative analysis, and one in a GRADE synthesis. The scientific evidence is insufficient to determine whether the accuracy of single reading + CAD is at least equivalent to that obtained in standard practice, i.e. double reading where two breast radiologists independently read the mammographic images.