Is mammary output capacity limiting to lactational performance in mice?
Using lactation in mice as a model, we sought to determine whether ceilings on sustained energy expenditure reside in the capacities of energy-acquiring and input organs (such as the intestine) or of energy-expending and output organs (such as the mammary glands). To distinguish between these possibilities experimentally, we surgically varied the teat number of lactating mother mice while simultaneously varying their litter size. The energy burden on each teat (i.e. the pup/teat ratio) could thus be varied independently of the energy burden (i.e. litter size) on the mother herself or on her intestine. At each teat number, pup mass proved to be maximal at intermediate litter sizes. At a given pup/teat ratio, mothers with five teats weaned pups no larger than the pups of normal (10-teat) mothers, even though the total energy burden on the former mothers was only half as large. Mothers with only two teats could not wean any pups. Litter size controlled maternal food intake, which in turn controlled intestinal mass and nutrient uptake capacity. Disproportionately high food intake for the smallest litters appears to reflect capital start-up costs of lactation. Pup mass is evidently limited by inadequate suckling stimulation of mammary glands.


Overview of the publications of rheumatologists after the millennium.
Scientific progress can be directly measured by the growth of the scientific archive. To our knowledge, there has never been a systematic evaluation of the number and properties of the publications of rheumatologists. In this paper, we aimed to evaluate the properties of these papers. All papers that were published between 2000 and 2011 by at least one rheumatologist author and that were indexed in Science Citation Index Expanded were evaluated. For this purpose, we used the ISI web of knowledge web of science software. Original articles were the most common document type. Most of the papers were published in rheumatology journals. From the point of the number of published papers, the top 3 countries were United States, England, and Germany; the top 3 journals were Annals of Rheumatic Diseases, Rheumatology, and Arthritis and Rheumatism, and the top 3 institutes are Harvard University, Leiden University, and The Imperial College of Science, Technology. Moreover, the publications of rheumatologists increased annually between 2000 and 2011. This paper is an overview of the publications of rheumatologists between 2000 and 2011. The results indicate an increasing trend of scientific productivity for rheumatologists. The outcomes about countries, institutions, and journals are in concordance with the general medical publication trends. More than half of the publications written by rheumatologist were published by rheumatology journals. We consider only quantitative data about publications; new research is required to qualitatively evaluate the data.


Gramene: A Resource for Comparative Analysis of Plants Genomes and Pathways.
Gramene is an integrated bioinformatics resource for accessing, visualizing, and comparing plant genomes and biological pathways. Originally targeting grasses, Gramene has grown to host annotations for over 90 plant genomes including agronomically important cereals (e.g., maize, sorghum, wheat, teff), fruits and vegetables (e.g., apple, watermelon, clementine, tomato, cassava), specialty crops (e.g., coffee, olive tree, pistachio, almond), and plants of special or emerging interest (e.g., cotton, tobacco, cannabis, or hemp). For some species, the resource includes multiple varieties of the same species, which has paved the road for the creation of species-specific pan-genome browsers. The resource also features plant research models, including Arabidopsis and C4 warm-season grasses and brassicas, as well as other species that fill phylogenetic gaps for plant evolution studies. Its strength derives from the application of a phylogenetic framework for genome comparison and the use of ontologies to integrate structural and functional annotation data. This chapter outlines system requirements for end-users and database hosting, data types and basic navigation within Gramene, and provides examples of how to (1) explore Gramene's search results, (2) explore gene-centric comparative genomics data visualizations in Gramene, and (3) explore genetic variation associated with a gene locus. This is the first publication describing in detail Gramene's integrated search interface-intended to provide a simplified entry portal for the resource's main data categories (genomic location, phylogeny, gene expression, pathways, and external references) to the most complete and up-to-date set of plant genome and pathway annotations.


Extensive degradation of RNA precursors by the exosome in wild-type cells.
The exosome is a complex involved in the maturation of rRNA and sn-snoRNA, in the degradation of short-lived noncoding RNAs, and in the quality control of RNAs produced in mutants. It contains two catalytic subunits, Rrp6p and Dis3p, whose specific functions are not fully understood. We analyzed the transcriptome of combinations of Rrp6p and Dis3p catalytic mutants by high-resolution tiling arrays. We show that Dis3p and Rrp6p have both overlapping and specific roles in degrading distinct classes of substrates. We found that transcripts derived from more than half of intron-containing genes are degraded before splicing. Surprisingly, we also show that the exosome degrades large amounts of tRNA precursors despite the absence of processing defects. These results underscore the notion that large amounts of RNAs produced in wild-type cells are discarded before entering functional pathways and suggest that kinetic competition with degradation proofreads the efficiency and accuracy of processing.


Hepatoregenerative role of bone morphogenetic protein-9.
Bone morphogenetic protein-9 (BMP-9) is a member of the transforming growth factor beta (TGF-β) superfamily of cytokines, which regulate cell growth and differentiation during embryogenesis. Apart of that, the hypoglycemic potential of BMP-9 is of great interest. It has been confirmed that BMP-9, like insulin, improves glycemia in diabetic mice and regulates directional glucose metabolism in hepatocytes; therefore it is proposed to be a candidate hepatic insulin-sensitizing substance (HISS). In liver fibrosis, due to the portocaval shunt, insulin bypasses the organ and the liver undergoes atrophy. Parenteral administration of insulin reverses atrophy by stimulating mitogenic activity of the hepatocytes. Because BMP-9 has a signaling pathway similar to other BMPs and insulin, it is to be expected that BMP-9 has a certain regenerative role in the liver, supporting the above-mentioned is evidence of BMP-9 expression in Dissè's spaces and BMP-7's mitogenic activity in mucosal cells. However, further studies are needed to confirm the possible regenerative role of BMP-9.


Epithelial barrier formation by airway basal cells.
Epithelial shedding processes in airway inflammation and defence may produce damaged areas where basal cells are the main remaining epithelial cell type. The present study examines the capacity of basal cells to form an epithelial barrier structure after loss of columnar epithelial cells. A technique was developed which allows selective removal of columnar epithelial cells from isolated airways. A drop of tissue adhesive glue was applied on the mucosal surface shortly after excision of guinea pig trachea and human bronchus. Gentle removal of the glue, together with attached columnar cells, left a single layer of cobbled, solitary basal cells. The tissue was kept in culture media. Morphological changes of the basal cells were monitored by immuno-histochemistry and scanning and transmission electron microscopy at several time points. After 20 minutes the basal cells had undergone extensive flattening and established contact with each other. The basement membrane thus became covered by a poorly differentiated epithelium in both guinea pig and human airways. Abundant interdigitating cytoplasmic protrusions were observed at cell borders. Basal cells promptly flatten out to cover the basement membrane at loss of neighbouring columnar cells. These data may explain why the epithelial barrier function may be uncompromised in desquamative airway diseases. Furthermore, they suggest the possibility that sacrificial release of columnar epithelial cells and prompt creation of a barrier structure constitute important roles of basal cells in airway defence against severe insults.


Computational approaches to cell-nanomaterial interactions: keeping balance between therapeutic efficiency and cytotoxicity.
Owing to their unique properties, nanomaterials have been widely used in biomedicine since they have obvious inherent advantages over traditional ones. However, nanomaterials may also cause dysfunction in proteins, genes and cells, resulting in cytotoxic and genotoxic responses. Recently, more and more attention has been paid to these potential toxicities of nanomaterials, especially to the risks of nanomaterials to human health and safety. Therefore, when using nanomaterials for biomedical applications, it is of great importance to keep the balance between therapeutic efficiency and cytotoxicity (i.e., increase the therapeutic efficiency as well as decrease the potential toxicity). This requires a deeper understanding of the interactions between various types of nanomaterials and biological systems at the nano/bio interface. In this review, from the point of view of theoretical researchers, we will present the current status regarding the physical mechanism of cytotoxicity caused by nanomaterials, mainly based on recent simulation results. In addition, the strategies for minimizing the nanotoxicity naturally and artificially will also be discussed in detail. Furthermore, we should notice that toxicity is not always bad for clinical use since causing the death of specific cells is the main way of treating disease. Enhancing the targeting ability of nanomaterials to diseased cells and minimizing their side effects on normal cells will always be hugely challenging issues in nanomedicine. By combining the latest computational studies with some experimental verifications, we will provide special insights into recent advances regarding these problems, especially for the design of novel environment-responsive nanomaterials.


Antisense oligodeoxynucleotides to inducible NO synthase rescue epithelial cells from oxidative stress injury.
Until recently, the lack of specific inhibitors of various forms of nitric oxide synthase (NOS) hampered a stringent evaluation of the role played by inducible NOS (iNOS) in cell damage. Phosphorothioate derivatives of iNOS antisense and control sense or scrambled oligodeoxynucleotides (S-ODNs) were synthesized, and their effect on epithelial cell viability was examined under oxidant stress. Exposure of BSC-1 kidney tubular epithelial cells to H2O2 resulted in elevation of NO release, accompanied by a significant decrease in the population of viable cells (from 97.4 +/- 1.7% to 72.4 +/- 2.4% population). Nitrite production by BSC-1 cells exposed to H2O2 increased almost 10-fold compared with control. Pretreatment of the cells with 10 microM antisense ODNs significantly blunted this response, whereas sense or scrambled ODNs did not modify it. Pretreatment of BSC-1 cells with 10 microM antisense ODNs virtually prevented lethal cell damage in response to H2O2, whereas sense ODNs were ineffective. Lipopolysaccharide induction of iNOS, also preventable by the antisense construct, resulted in a lesser compromise to cell viability. Immunocytochemistry of iNOS in cells pretreated with antisense ODNs showed minimal cytoplasmic staining, as opposed to the untreated or sense ODN-treated positively stained cells. Staining with antibodies to nitrotyrosine was conspicuous in stressed cells but undetectable in antisense ODN-treated cells. In conclusion, oxidant stress is accompanied by the induction of iNOS, increased production of NO, and impaired cell viability; selective inhibition of iNOS using the designed antisense ODNs dramatically improved BSC-1 cell viability after oxidant stress.


Granulocytic sarcoma, an undiagnosed leukemia, initially manifested as paralysis.
Granulocytic sarcoma (GS) may be a presenting sign of myelogenous leukemia. Occasionally, an extramedullary neoplasm composed from myelocytic precursor cells occurs in patients without evidence of leukemia. Rarely, undiagnosed leukemia occurs initially manifesting with paralysis to spinal cord GS. We present a case report of 20-year-old girl with an undiagnosed leukemia, initially manifesting as paralysis. En bloc spondylectomy with chemotherapy postoperatively constituted the treatment of choice for this tumor. After two courses of chemotherapy, the patient made a good postoperative recovery with notable bilateral lower extremity improvement.


Src Dependent Pancreatic Acinar Injury Can Be Initiated Independent of an Increase in Cytosolic Calcium.
Several deleterious intra-acinar phenomena are simultaneously triggered on initiating acute pancreatitis. These culminate in acinar injury or inflammatory mediator generation in vitro and parenchymal damage in vivo. Supraphysiologic caerulein is one such initiator which simultaneously activates numerous signaling pathways including non-receptor tyrosine kinases such as of the Src family. It also causes a sustained increase in cytosolic calcium- a player thought to be crucial in regulating deleterious phenomena. We have shown Src to be involved in caerulein induced actin remodeling, and caerulein induced changes in the Golgi and post-Golgi trafficking to be involved in trypsinogen activation, which initiates acinar cell injury. However, it remains unclear whether an increase in cytosolic calcium is necessary to initiate acinar injury or if injury can be initiated at basal cytosolic calcium levels by an alternate pathway. To study the interplay between tyrosine kinase signaling and calcium, we treated mouse pancreatic acinar cells with the tyrosine phosphatase inhibitor pervanadate. We studied the effect of the clinically used Src inhibitor Dasatinib (BMS-354825) on pervanadate or caerulein induced changes in Src activation, trypsinogen activation, cell injury, upstream cytosolic calcium, actin and Golgi morphology. Pervanadate, like supraphysiologic caerulein, induced Src activation, redistribution of the F-actin from its normal location in the sub-apical area to the basolateral areas, and caused antegrade fragmentation of the Golgi. These changes, like those induced by supraphysiologic caerulein, were associated with trypsinogen activation and acinar injury, all of which were prevented by Dasatinib. Interestingly, however, pervanadate did not cause an increase in cytosolic calcium, and the caerulein induced increase in cytosolic calcium was not affected by Dasatinib. These findings suggest that intra-acinar deleterious phenomena may be initiated independent of an increase in cytosolic calcium. Other players resulting in acinar injury along with the Src family of tyrosine kinases remain to be explored.


Laparoscopic Lavage Is Feasible and Safe for the Treatment of Perforated Diverticulitis With Purulent Peritonitis: The First Results From the Randomized Controlled Trial DILALA.
To evaluate short-term outcomes of a new treatment for perforated diverticulitis with purulent peritonitis in a randomized controlled trial. Perforated diverticulitis with purulent peritonitis (Hinchey III) has traditionally been treated with surgery including colon resection and stoma (Hartmann procedure) with considerable postoperative morbidity and mortality. Laparoscopic lavage has been suggested as a less invasive surgical treatment. Laparoscopic lavage was compared with colon resection and stoma in a randomized controlled multicenter trial, DILALA (ISRCTN82208287). Initial diagnostic laparoscopy showing Hinchey III was followed by randomization. Clinical data was collected up to 12 weeks postoperatively. Eighty-three patients were randomized, out of whom 39 patients in laparoscopic lavage and 36 patients in the Hartmann procedure groups were available for analysis. Morbidity and mortality after laparoscopic lavage did not differ when compared with the Hartmann procedure. Laparoscopic lavage resulted in shorter operating time, shorter time in the recovery unit, and shorter hospital stay. In this trial, laparoscopic lavage as treatment for patients with perforated diverticulitis Hinchey III was feasible and safe in the short-term.


Lipoma with dumb-bell extradural extension through the intervertebral foramen into the spinal canal. Case report.
The authors present the case of a 60-year-old woman with a neck lipoma that developed dumb-bell extradural extension, causing radiculopathy. To the best of the authors' knowledge, this is the first report of a lipoma originating in the neck with dumb-bell extradural extension through the intervertebral foramen and into the spinal canal. The lipoma was first excised from the foramen via a posterior approach to allow decompression of the nerve roots. The remaining lipomatous tissue was then resected via an anterior approach to avoid the region around the vertebral artery.


Genotoxicity and toxicity assessment in urban hydrographic basins.
The genotoxicity and cytotoxicity of water in small urban basins was evaluated by the Salmonella/microsome assay and micronucleus test in V79 cells. The results showed that the cytotoxic effect was the most significant response in areas with medium to heavy urban occupation for both assays evaluated. Water samples from these areas include different concentrations of chloroform, bromodichloromethane, toluene, ethylbenzene, m,p-xylene and 1,4-dichlorobenzene. As to genotoxic damage, the presence of mainly direct-acting frameshift mutagens was detected in areas with less urban concentration and showed genotoxic activity in V79 cells in more heavily urbanized areas. Water organic extracts, evaluated using a microsuspension procedure, showed frameshift mutagenic activity in the presence of hepatic metabolization that increased as the population density grow. Chronic toxicity studies of sediment samples with the microcrustacean Daphnia magna showed that, while survival was not highly affected, reproductive inhibition was found in 92% of the observations. A retrospective diagnosis of water quality using traditional physicochemical parameters that defined the differential contribution of urban wastes at the three sites was associated with the biological assays. It became clear that the biological assays were of significant benefit in the diagnosis of risks of contamination of hydrographic basins by pollutants from urban non-point sources.


Protein tyrosine phosphatase non-receptor type 22 (PTPN22) gene R620W variant and sporadic idiopathic hypoparathyroidism in Asian Indians.
Recently, a gain of function variant C1858T of the lymphoid-specific protein tyrosine phosphatase non-receptor (LYP, PTPN22) gene has been reported to be associated with several autoimmune disorders including Graves' disease, type 1 diabetes, rheumatoid arthritis and vitiligo. The present study was carried out in 80 patients with sporadic idiopathic hypoparathyroidism (SIH) [43 males and 37 females, mean +/- SD age and duration of symptoms 32.5 +/- 14.1 years and 6.7 +/- 7.2 years (range 1 day to 35 years), respectively] and 193 healthy controls (male : female ratio 91:102, mean +/- SD age, 43.1 +/- 11.6 years) to assess association of 1858T allele with the disease. Polymerase chain reaction-restriction fragment length polymorphism analysis was performed to genotype C1858T variant. The frequency of occurrence of 1858T allele was 4/160 (2.5%) in SIH and 5/386 (1.3%) in the control alleles (odds ratio 1.95, 95% CI 0.51-7.37). Thus, the present study reveals that 1858T allele is rare (1.3%) in Asian Indians. The trend of higher prevalence of 1858T allele in patients with SIH needs to be studied further in other population with higher rate of the allele to support the autoimmune basis of the disease.


Detecting Protein-Glycolipid Interactions Using Glycomicelles and CaR-ESI-MS.
This study reports on the use of the catch-and-release electrospray ionization mass spectrometry (CaR-ESI-MS) assay, combined with glycomicelles, as a method for detecting specific interactions between water-soluble proteins and glycolipids (GLs) in aqueous solution. The B subunit homopentamers of cholera toxin (CTB5) and Shiga toxin type 1 B (Stx1B5) and the gangliosides GM1, GM2, GM3, GD1a, GD1b, GT1b, and GD2 served as model systems for this study. The CTB5 exhibits broad specificity for gangliosides and binds to GM1, GM2, GM3, GD1a, GD1b, and GT1b; Stx1B5 does not recognize gangliosides. The CaR-ESI-MS assay was used to analyze solutions of CTB5 or Stx1B5 and individual gangliosides (GM1, GM2, GM3, GD1a, GD1b, GT1b, and GD2) or mixtures thereof. The high affinity interaction of CTB5 with GM1 was successfully detected. However, the apparent affinity, as determined from the mass spectra, is significantly lower than that of the corresponding pentasaccharide or when GM1 is presented in model membranes such as nanodiscs. Interactions between CTB5 and the low affinity gangliosides GD1a, GD1b, and GT1b, as well as GD2, which served as a negative control, were detected; no binding of CTB5 to GM2 or GM3 was observed. The CaR-ESI-MS results obtained for Stx1B5 reveal that nonspecific protein-ganglioside binding can occur during the ESI process, although the extent of binding varies between gangliosides. Consequently, interactions detected for CTB5 with GD1a, GD1b, and GT1b are likely nonspecific in origin. Taken together, these results reveal that the CaR-ESI-MS/glycomicelle approach for detecting protein-GL interactions is prone to false positives and false negatives and must be used with caution. Graphical Abstract <!-- [INSERT GRAPHICAL ABSTRACT TEXT HERE] -->.


MicroRNA-338-5p modulates pulmonary hypertension-like injuries caused by SO2, NO2 and PM2.5 co-exposure through targeting the HIF-1α/Fhl-1 pathway.
The role of ambient air pollution is considered to be important in the development of chronic obstructive pulmonary disease (COPD), and pulmonary hypertension (PH) is a common clinical manifestation of COPD. However, many studies have mainly focused on the adverse health effects of a single air pollutant, ignoring the combined toxicity of multiple pollutants. In the present study, we co-exposed mice to coal-burning air pollutants (SO2, NO2 and PM2.5), and confirmed PH-like injury occurrence by airflow limitation, marked abnormal endothelin-1 (ET-1) and endothelial nitric oxide synthase (eNOS) expression, and histopathological and ultrastructural alteration. Global microRNA (miRNA) arrays identified three significantly changed miRNAs homologous with humans (miR-338-5p, miR-450b-3p and miR-142-5p), and we targeted miR-338-5p based on real-time reverse transcription-PCR (RT-PCR) validation. Furthermore, bioinformatic and dual-luciferase reporter gene analyses indicated that miR-338-5p bound to 3'-UTR of hypoxia-inducible factor 1α (HIF-1α) mRNA and down-regulation of miR-338-5p led to the increased expression of HIF-1α and its related gene four-and-a-half LIM (Lin-11, Isl-1 and Mec-3) domain 1 (Fhl-1) and contributed to PH. This study provides evidence for the role of miRNAs in PH through targeting HIF-1α/Fhl-1 pathway after air pollutants co-exposure and implies new insights into the molecular markers for COPD caused by air pollution.


Nanoparticle design strategies for enhanced anticancer therapy by exploiting the tumour microenvironment.
Nanovehicles can efficiently carry and deliver anticancer agents to tumour sites. Compared with normal tissue, the tumour microenvironment has some unique properties, such as vascular abnormalities, hypoxia and acidic pH. There are many types of cells, including tumour cells, macrophages, immune and fibroblast cells, fed by defective blood vessels in the solid tumour. Exploiting the tumour microenvironment can benefit the design of nanoparticles for enhanced therapeutic effectiveness. In this review article, we summarized the recent progress in various nanoformulations for cancer therapy, with a special emphasis on tumour microenvironment stimuli-responsive ones. Numerous tumour microenvironment modulation strategies with promising cancer therapeutic efficacy have also been highlighted. Future challenges and opportunities of design consideration are also discussed in detail. We believe that these tumour microenvironment modulation strategies offer a good chance for the practical translation of nanoparticle formulas into clinic.


Evaluation of exposure to contaminated drinking water and specific birth defects and childhood cancers at Marine Corps Base Camp Lejeune, North Carolina: a case-control study.
Drinking water supplies at Marine Corps Base Camp Lejeune were contaminated with trichloroethylene, tetrachloroethylene, benzene, vinyl chloride and trans-1,2-dichloroethylene during 1968 through 1985. We conducted a case control study to determine if children born during 1968-1985 to mothers with residential exposure to contaminated drinking water at Camp Lejeune during pregnancy were more likely to have childhood hematopoietic cancers, neural tube defects (NTDs), or oral clefts. For cancers, exposures during the first year of life were also evaluated. Cases and controls were identified through a survey of parents residing on base during pregnancy and confirmed by medical records. Controls were randomly sampled from surveyed participants who had a live birth without a major birth defect or childhood cancer. Groundwater contaminant fate and transport and distribution system models provided estimates of monthly levels of drinking water contaminants at mothers' residences. Magnitude of odds ratios (ORs) was used to assess associations. Confidence intervals (CIs) were used to indicate precision of ORs. We evaluated parental characteristics and pregnancy history to assess potential confounding. Confounding was negligible so unadjusted results were presented. For NTDs and average 1st trimester exposures, ORs for any benzene exposure and for trichloroethylene above 5 parts per billion were 4.1 (95% CI: 1.4-12.0) and 2.4 (95% CI: 0.6-9.6), respectively. For trichloroethylene, a monotonic exposure response relationship was observed. For childhood cancers and average 1st trimester exposures, ORs for any tetrachloroethylene exposure and any vinyl chloride exposure were 1.6 (95% CI: 0.5-4.8), and 1.6 (95% CI: 0.5-4.7), respectively. The study found no evidence suggesting any other associations between outcomes and exposures. Although CIs were wide, ORs suggested associations between drinking water contaminants and NTDs. ORs suggested weaker associations with childhood hematopoietic cancers.


Constrained handgrip force decreases upper extremity muscle activation and arm strength.
Many industrial tasks require repetitive shoulder exertions to be performed with concurrent physical and mental demands. The highly mobile nature of the shoulder predisposes it to injury. The purpose of this study was to determine the effects of simultaneous gripping, at a specified magnitude, on muscle activity and maximal arm force in various directions. Ten female subjects performed maximal arm exertions at two different heights and five directions using both specified (30% maximum voluntary grip) and preferred (self-selected) grip forces. Electromyography was recorded from eight muscles of the right upper extremity. The preferred grip condition produced grip forces that were dependent on the combination of arm height and force direction and were significantly greater (arm force down), lower (to left, up and push forward), or similar to the specified grip condition. Regardless of the magnitude of the preferred grip force, specifying the grip resulted in decreased maximal arm strength (by 18-25%) and muscle activity (by 15-30%) in all conditions, indicating an interfering effect when the grip force was specified by visual target force-matching. Task constraints, such as specific gripping demands, may decrease peak force levels attainable and alter muscle activity. Depending on the nature of task, the amount of relative demand may differ, which should be considered when determining safety thresholds.


Parallel secretory pathways to the cell surface in yeast.
Saccharomyces cerevisiae mutants that have a post-Golgi block in the exocytic pathway accumulate 100-nm vesicles carrying secretory enzymes as well as plasma membrane and cell-wall components. We have separated the vesicle markers into two groups by equilibrium isodensity centrifugation. The major population of vesicles contains Bg12p, an endoglucanase destined to be a cell-wall component, as well as Pma1p, the major plasma membrane ATPase. In addition, Snc1p, a synaptobrevin homologue, copurifies with these vesicles. Another vesicle population contains the periplasmic enzymes invertase and acid phosphatase. Both vesicle populations also contain exoglucanase activity; the major exoglucanase normally secreted from the cell, encoded by EXG1, is carried in the population containing periplasmic enzymes. Electron microscopy shows that both vesicle groups have an average diameter of 100 nm. The late secretory mutants sec1, sec4, and sec6 accumulate both vesicle populations, while neither is detected in wild-type cells, early sec mutants, or a sec13 sec6 double mutant. Moreover, a block in endocytosis does not prevent the accumulation of either vesicle species in an end4 sec6 double mutant, further indicating that both populations are of exocytic origin. The accumulation of two populations of late secretory vesicles indicates the existence of two parallel routes from the Golgi to the plasma membrane.


Potential of milk mid-infrared spectra to predict nitrogen use efficiency of individual dairy cows in early lactation.
Improving nitrogen use efficiency (NUE) at both the individual cow and the herd level has become a key target in dairy production systems, for both environmental and economic reasons. Cost-effective and large-scale phenotyping methods are required to improve NUE through genetic selection and by feeding and management strategies. The aim of this study was to evaluate the possibility of using mid-infrared (MIR) spectra of milk to predict individual dairy cow NUE during early lactation. Data were collected from 129 Holstein cows, from calving until 50 d in milk, in 3 research herds (Denmark, Ireland, and the UK). In 2 of the herds, diets were designed to challenge cows metabolically, whereas a diet reflecting local management practices was offered in the third herd. Nitrogen intake (kg/d) and nitrogen excreted in milk (kg/d) were calculated daily. Nitrogen use efficiency was calculated as the ratio between nitrogen in milk and nitrogen intake, and expressed as a percentage. Individual daily values for NUE ranged from 9.7 to 81.7%, with an average of 36.9% and standard deviation of 10.4%. Milk MIR spectra were recorded twice weekly and were standardized into a common format to avoid bias between apparatus or sampling periods. Regression models predicting NUE using milk MIR spectra were developed on 1,034 observations using partial least squares or support vector machines regression methods. The models were then evaluated through (1) a cross-validation using 10 subsets, (2) a cow validation excluding 25% of the cows to be used as a validation set, and (3) a diet validation excluding each of the diets one by one to be used as validation sets. The best statistical performances were obtained when using the support vector machines method. Inclusion of milk yield and lactation number as predictors, in combination with the spectra, also improved the calibration. In cross-validation, the best model predicted NUE with a coefficient of determination of cross-validation of 0.74 and a relative error of 14%, which is suitable to discriminate between low- and high-NUE cows. When performing the cow validation, the relative error remained at 14%, and during the diet validation the relative error ranged from 12 to 34%. In the diet validation, the models showed a lack of robustness, demonstrating difficulties in predicting NUE for diets and for samples that were not represented in the calibration data set. Hence, a need exists to integrate more data in the models to cover a maximum of variability regarding breeds, diets, lactation stages, management practices, seasons, MIR instruments, and geographic regions. Although the model needs to be validated and improved for use in routine conditions, these preliminary results showed that it was possible to obtain information on NUE through milk MIR spectra. This could potentially allow large-scale predictions to aid both further genetic and genomic studies, and the development of farm management tools.


Spectrum of Diagnosis and Disposition of Patients Referred to a Pediatric Orthopaedic Center for a Diagnosis of Intoeing.
Orthopaedic surgeons frequently evaluate otherwise healthy children for concern of intoed gait. Intoeing in otherwise healthy young children due to metatarsus adductus, internal tibial torsion, and increased femoral anteversion do not typically require orthopaedic treatment. This study reviewed the actual diagnosis, management, and disposition of patients referred to a pediatric orthopaedic specialty hospital for a diagnosis of intoeing; the efficacy of an Advanced Practice Provider (APP) assessment program to screen and triage patients with a primary complaint of intoeing; and parental satisfaction with that program. We established an "Intoeing Clinic" conducted by APPs to conduct initial evaluations of patients referred for a diagnosis of intoeing meeting-specific criteria, including (1) a referring provider's diagnosis of "intoeing"; (2) the patient was under the age of 9 years; and (3) there was no suggestion of comorbidity in the information provided by the referring provider to imply a diagnosis other than "benign" intoeing. Under pediatric orthopaedic surgeon "on-call" supervision, APPs were authorized to perform clinical assessments supplemented by radiographs and laboratory investigations as deemed necessary. We performed an Institutional Review Board-approved, retrospective medical record review of all patients appointed to our Intoeing Clinic over a 30-month period (March 2010 to September 2013). About 95% of 926 patients appointed to APP Intoeing Clinic were confirmed to have a diagnosis of "benign" intoeing or a similar "benign" diagnosis; 5% of these patients requested a reevaluation for the same concern. Approximately 5% were determined to have a nonbenign diagnosis, either known to the family/provider, but not conveyed at the time of referral (4%), or identified at our institution (1%). Two patients (0.2%) were determined at follow-up examination to have a neurological abnormality at the subsequent examination. An "Intoeing Clinic" staffed by experienced Advanced Pediatric Practitioners or equivalent, with appropriate orthopaedic surgeon availability for consultation can be an effective and efficient method of evaluating patients referred for a diagnosis of "intoeing." Level IV-case series.


Prenatal urban traffic noise exposure impairs spatial learning and memory and reduces glucocorticoid receptor expression in the hippocampus of male rat offspring.
Exposure to noise stress during early life may permanently affect the structure and function of the central nervous system. The aim of this study was to evaluate the effects of prenatal exposure to urban traffic noise on the spatial learning and memory of the rats' offspring and the expression of glucocorticoid receptors (GRs) in their hippocampi. Three g\roups of pregnant rats were exposed to recorded urban traffic noise for 1, 2 or 4 h/day during the last week of pregnancy. At the age of 45 days, their male offspring were introduced to the Morris water maze (MWM) for assessment of spatial learning and memory. The corticosterone levels were measured in the offspring's sera by radioimmunoassay, and the relative expression of glucocorticoid and mineralocorticoid receptors (MRs) in their hippocampi was evaluated via RT-PCR. Facing urban traffic noise for 2 and 4 h/day during the third trimester of pregnancy caused the offspring to spend more time and to travel a larger distance than the controls to find the target platform. Analogously, these two groups were inferior to their control counterparts in the probe test. Also, prenatal noise stress elevated the corticosterone concentration in the sera of the rats' offspring and dose-dependently decreased the relative expression of the mRNA of both GRs and MRs in their hippocampi. Urban traffic noise exposure during the last trimester of pregnancy impairs spatial learning and memory of rat offspring and reduces GRs and MRs gene expression in the hippocampus.


Use of porous tantalum metaphyseal cones for severe tibial bone loss during revision total knee replacement. Surgical technique.
The best treatment method for large tibial bone defects during revision knee replacement has not been established. The purpose of this study was to determine the initial results obtained with a unique reconstructive implant, the porous tantalum metaphyseal cone, designed as an alternative treatment for severe tibial bone loss following total knee arthroplasty. Porous tantalum metaphyseal cones were implanted during fifteen revision total knee replacements in eight women and seven men who had an average age of 68.1 years at the time of the procedure. The patients had had an average of 3.5 prior total knee replacements. According to the Anderson Orthopaedic Research Institute bone defect classification, eight knees had a Type-3 defect and seven knees had a Type-2B bone defect. All patients were followed clinically and radiographically. The patients were followed for an average of thirty-four months (range, twenty-four to forty-seven months). Overall, the average Knee Society clinical scores improved from 52 points preoperatively to 85 points at the time of the final follow-up. At the final follow-up evaluation, all fifteen porous metaphyseal cones showed evidence of osseointegration with reactive osseous trabeculation at points of contact with the tibia. There was no evidence of loosening or migration of any of these tibial reconstructions at the time of final follow-up. At the time of short-term follow-up, the porous tantalum metaphyseal tibial cones effectively provided structural support for the tibial implants in this series. The potential for long-term biologic fixation may provide durability for these tibial reconstructions. Long-term follow-up and comparison with alternative reconstructive techniques will be required to evaluate the true effectiveness of this treatment approach.


Reference values of impulse oscillometry and its utility in the diagnosis of asthma in young Korean children.
The aims of this study were (1) to determine the reference values for impulse oscillometry (IOS) and (2) to apply them to the evaluation of asthma in the general population of young Korean children. We performed a questionnaire survey and IOS measurements in 390 children aged 3-7 years in Seoul and Gyeonggi province, Korea, from July to August 2010. IOS measurements included respiratory resistance (Rrs) and respiratory reactance (Xrs) at 5, 10, 15, 20, 25, and 35 Hz, respiratory impedance (Zrs), and resonance frequency (RF) before and 15 min after inhalation of 200 μg salbutamol. To determine the reference values for IOS, 161 children defined as healthy controls were assessed. The IOS measurements were presented as means and standard deviations. The reference equations for IOS variables were determined by multiple linear regression analysis taking into account their height, weight, and age (R5 = 2.242 - 0.008 × height (cm) - 0.005 × age (months), coefficients of determination (R(2)) = 0.213). Height had the greatest correlation with IOS variables, similar to previous studies. Positive airway obstruction was defined as R5 greater than the 95th percentile of predicted R5 from the reference equation. There was a higher percentage of children with positive airway obstruction in children with asthma than in healthy controls (27.3% vs. 6.2%). Multivariate logistic regression analysis indicated that positive airway obstruction was a significant risk factor for the diagnosis of asthma (adjusted odds ratio (aOR), 6.245; 95% confidence interval (CI), 2.270-17.175). This study provided reference values for IOS in young Korean children and applied the reference values to evaluate children with asthma. We suggest the 95th percentile of predicted R5 as a cut-off value for positive airway obstruction, which may increase the risk for diagnosis of asthma.


Discovery of a polyomavirus in European badgers (Meles meles) and the evolution of host range in the family Polyomaviridae.
Polyomaviruses infect a diverse range of mammalian and avian hosts, and are associated with a variety of symptoms. However, it is unknown whether the viruses are found in all mammalian families and the evolutionary history of the polyomaviruses is still unclear. Here, we report the discovery of a novel polyomavirus in the European badger (Meles meles), which to our knowledge represents the first polyomavirus to be characterized in the family Mustelidae, and within a European carnivoran. Although the virus was discovered serendipitously in the supernatant of a cell culture inoculated with badger material, we subsequently confirmed its presence in wild badgers. The European badger polyomavirus was tentatively named Meles meles polyomavirus 1 (MmelPyV1). The genome is 5187 bp long and encodes proteins typical of polyomaviruses. Phylogenetic analyses including all known polyomavirus genomes consistently group MmelPyV1 with California sea lion polyomavirus 1 across all regions of the genome. Further evolutionary analyses revealed phylogenetic discordance amongst polyomavirus genome regions, possibly arising from evolutionary rate heterogeneity, and a complex association between polyomavirus phylogeny and host taxonomic groups.


Polymorphism in the 5'-flanking region of the human insulin gene and its possible relation to type 2 diabetes.
The arrangement of the human insulin gene in DNA from 87 individuals was analyzed by the Southern blot hybridization technique with a cloned genomic human insulin probe. Insertions of 1.5 to 3.4 kilobase pairs in the 5'-flanking region of the gene were found in DNA from 38 individuals. These insertions occurred within 1.3 kilobase pairs of the transcription initiation site. In contrast, no insertions were observed in the region 3' to the coding sequence. The prevalence of these insertions in type 2 diabetes was significantly greater than in the other groups (P less than .001). The limitation of this striking length polymorphism to a potential promoter region suggests that these insertions may play a role in insulin gene expression.


Prevention and treatment of alcohol-related problems: research opportunities. Committee to Identify Research Opportunities in the Prevention and Treatment of Alcohol-Related Problems.
This article summarizes the major themes, conclusions and recommendations of a 2-year study conducted for the National Institute on Alcohol Abuse and Alcoholism by the Institute of Medicine. The study is devoted to the identification of research opportunities that will improve knowledge about the prevention and treatment of alcohol problems. In the first part of the report, the study committee examines the social and personal aspects of alcohol-related problems toward which prevention efforts are directed; delineates the features of a public health orientation that it deems most appropriate for the prevention task; discusses individual vulnerability to alcohol misuse; and reviews genetic, developmental and social learning perspectives on prevention. In the second part of the report, which is devoted to treatment research, the committee considers the underlying philosophical issues as well as the formidable methodological problems in conducting treatment research. Central to this is a broad review of promising treatment modalities and the research needed for developing effective patient-treatment matching schemes. In the final part of the report, the committee concludes that cooperative multisite research efforts are indispensable to the implementation of the research directions it recommends.


Intraluminal crystalloids in whole-organ sections of prostate.
Demonstration of intraluminal crystalloids and mucin can aid in the diagnosis of prostatic adenocarcinoma. Crystalloids have been reported in 10% to 23% of prostatic adenocarcinomas. This incidence may not be accurate, however, because previous studies were based on specimens obtained by transurethral resection or transrectal biopsy. This study was based on the examination of 54 prostates (9 obtained from radical prostatectomies for prostate cancer and 45 from cystoprostatectomies for bladder carcinoma) processed by a whole-organ section method. Crystalloids were found in all nine prostatic carcinomas from radical prostatectomy specimens. Thirty-one of 45 specimens from cystoprostatectomies had single or multiple foci of adenocarcinoma; of these, 20 cases (64.5%) had crystalloids. Numbers of crystalloids varied not only from case to case but also from area to area within the same case. Crystalloids were occasionally identified in benign and dysplastic glands that were adjacent to areas of carcinoma. Cases without prostatic carcinoma and benign or dysplastic glands distant from areas of carcinoma did not contain crystalloids. Twenty-four prostatic adenocarcinomas had intraluminal mucin; mucin was not found in benign glands. In conclusion we found the incidence of crystalloids to be higher than that reported in previous studies, a difference we believe is because of sampling. We confirm previous observations that intraluminal crystalloids and mucin are helpful in diagnosing prostate adenocarcinoma.


Cerebellar agenesis revisited.
New clinical and employment information, together with over-looked previously published information, on a patient (H.C.) is reviewed. H.C., who died at the age of 76 in 1939, was found, by chance during anatomical dissection, to lack a cerebellum. This synthesis challenges an unusual and interesting account of cerebellar agenesis published in Brain in 1994 by Glickstein (see also Glickstein, 2006), in which the allegedly 'bogus' oral history of this individual's motor skills was held to have led to 'medical myth making'. Part of the burden of the 1994 paper was to show that 'cerebellar agenesis is always associated with profound motor deficits'. Glickstein therefore focussed on an apparent 'exception' to this conclusion, concerning the brain of a single case, H.C., who died 70 years ago, who 'had given rise to an oral tradition alleging that normal movement is possible despite total cerebellar agenesis'. Glickstein (1994) concludes 'despite an oral tradition to the contrary there is absolutely no evidence about the motor capacities of this man during his life'. Rather remarkably, an extensive history of this individual has become available, its significance becoming noted only this year; this complements and adds to a previous brief history published on H.C. (and not mentioned in the 1994 paper; see below). The new evidence includes the death certificate stating the man's occupation to have been 'manual labourer' with all the implications relevant to his supposed incapacity. The written historical record thus confronts the alleged 'myth'. It is interesting to note how medical records on an undoubtedly very ordinary citizen were recorded in London in the 1930s (before the NHS was set up in 1949) and how they could be made accessible to clinical colleagues in east London in the middle of World War II blitz bombing of the capital.


Burden of de novo malignancy in the liver transplant recipient.
Recipients of liver transplantation (LT) have a higher overall risk (2-3 times on average) of developing de novo malignancies than the general population, with standardized incidence ratios ranging from 1.0 for breast and prostate cancers to 3-4 for colon cancer and up to 12 for esophageal and oropharyngeal cancers. Aside from immunosuppression, other identified risk factors for de novo malignancies include the patient's age, a history of alcoholic liver disease or primary sclerosing cholangitis, smoking, and viral infections with oncogenic potential. Despite outcome studies showing that de novo malignancies are major causes of mortality and morbidity after LT, there are no guidelines for cancer surveillance protocols or immunosuppression protocols to lower the incidence of de novo cancers. Patient education, particularly for smoking cessation and excess sun avoidance, and regular clinical follow-up remain the standard of care. Further research in epidemiology, risk factors, and the effectiveness of screening and management protocols is needed to develop evidence-based guidelines for the prevention and treatment of de novo malignancies.


Dynamics of magnetization in hyperpolarized gas MRI of the lung.
The magnetization in hyperpolarized gas (HP) MRI is generated by laser polarization that is independent of the magnet and imaging process. As a consequence, there is no equilibrium magnetization during the image acquisition. The competing processes of gas inflow and depolarization of the spins lead to large changes in signal as one samples k-space. A model is developed of dynamic changes in polarization of hyperpolarized 3He during infusion and in vivo imaging of the lung and verified experimentally in a live guinea pig. Projection encoding is used to measure the view-to-view variation with temporal resolution < 4 ms. Large excitation angles effectively sample the magnetization in the early stages of inflow, highlighting larger airways, while smaller excitation angles produce images of the more distal spaces. The work provides a basis for pulse sequences designed to effectively exploit HP MRI in the lung.


Dietary combination of fish oil and taurine decreases fat accumulation and ameliorates blood glucose levels in type 2 diabetic/obese KK-A(y) mice.
n-3 Polyunsaturated fatty acids (PUFAs) such as eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), and taurine are functional compounds abundantly present in seafoods. In this study, we examined the combined effects of EPA- and DHA-rich fish oil and taurine on white adipose tissue (WAT) weight and blood glucose levels in diabetic/obese KK-A(y) mice. After a 4-wk administration of experimental diets (soybean oil or fish oil, supplemented with 0%, 2%, or 4% taurine), the increase in WAT weight of the mice fed the "fish oil + 4% taurine" diet was significantly suppressed compared to the "soybean oil + 4% taurine" and "fish oil only" diets. Serum triglycerides, free fatty acids, and total cholesterol levels decreased by fish oil administration. In addition, fish oil and taurine increased the activity of acyl-CoA oxidase, which is the rate-limiting enzyme of peroxisomal β-oxidation, increased in the liver of KK-A(y) mice. The activity of fatty acid synthase decreased by fish oil diets. Furthermore, blood glucose and insulin levels were significantly lower in the mice fed fish oil than in the soybean oil-fed mice. In fish oil + 4% taurine group, hyperglycemia and hyperinsulinemia were effectively improved in KK-A(y) mice compared to the fish oil only groups. In particular, the combination of fish oil and taurine enhanced the glucose transporter 4 (GLUT4) distribution in the plasma membrane of muscle tissue. These results suggest that EPA- and DHA-rich fish oil, especially in combination with taurine, exhibits preventive effects on WAT weight gain and hyperglycemia in diabetic/obese KK-A(y) mice.


Long term safety of sacral nerve modulation in medicare beneficiaries.
Sacral nerve stimulation (SNS) is FDA approved as second-line therapy for both urinary and bowel control. However, there is limited evidence regarding long term safety. We determined adverse events associated with SNS among Medicare beneficiaries. We used the 5% national random sample of Medicare claims for 2001-2011 to identify patients. Patients who underwent SNS implantation were identified with Current Procedure Terminology (CPT-4) codes. We determined safety of SNS using analysis of complication occurrences on day of surgery and during 5 years following initial procedure. SAS v9.3 statistical package was used. One thousand four hundred seventy-four patients underwent treatment with SNS in the 5% national sample of Medicare patients within the time period. Representative of real-world patients undergoing SNS surgery, comorbidities included hypertension (69.3%), diabetes (29.4%), chronic pulmonary disease (25.5%), hypothyroidism (25.2%), and depression (22.7%). Few complications occurred on day of surgery. At 90 days, 3.2% of patients had bowel complications, 2.0% urological, 9.4% infectious, and 1.5% stroke. Overall, bowel, neurological health event occurrences were consistent with prior year rates, while infectious events decreased. Of 206 patients who were followed for at least 5 years, 17.3% had devices removed and 11.3% replaced, with 26.1% having at least one of those, leaving 73.9% with original devices. Urological, infectious, and bowel complication occurrences were low after SNS among Medicare beneficiaries with multiple comorbidities. There were infrequent serious complications like hemorrhage and stroke postoperatively. Although SNS appears safe in this high-risk population, a comprehensive registry will ensure continuous safety.


A Population-based survey of the prevalence and types of glaucoma in Nigeria: results from the Nigeria National Blindness and Visual Impairment Survey.
Glaucoma is the leading cause of irreversible blindness worldwide. There tends to be a lower reporting of glaucoma in Africa compared to other blinding conditions in global burden data. Research findings of glaucoma in Nigeria will significantly increase our understanding of glaucoma in Nigeria, in people of the West African diaspora and similar population groups. We determined the prevalence and types of glaucoma in Nigeria from the Nigeria National Blindness and Visual Impairment cross-sectional Survey of adults aged ≥40 years. Multistage stratified cluster random sampling with probability-proportional-to-size procedures were used to select a nationally representative sample of 15,027 persons aged ≥40 years. Participants had logMAR visual acuity measurement, FDT visual function testing, autorefraction, A-scan biometry and optic disc assessment. Participants with visual acuity of worse than 6/12 or suspicious optic discs had detailed examination including Goldmann applanation tonometry, gonioscopy and fundus photography. Disc images were graded by Moorfields Eye Hospital Reading Centre. Glaucoma was defined using International Society of Geographical and Epidemiological Ophthalmology criteria; and classified into primary open-angle or primary angle-closure or secondary glaucoma. Diagnosis of glaucoma was based on ISGEO classification. The type of glaucoma was determined by gonioscopy. A total of 13,591 participants in 305 clusters were examined (response rate 90.4 %). Optic disc grading was available for 25,289 (93 %) eyes of 13,081 (96 %) participants. There were 682 participants with glaucoma; a prevalence of 5.02 % (95 % CI 4.60-5.47). Among those with definite primary glaucoma that had gonioscopy (n = 243), open-angle glaucoma was more common (86 %) than angle-closure glaucoma (14 %). 8 % of glaucoma was secondary with the commonest causes being couching (38 %), trauma (21 %) and uveitis (19 %). Only 5.6 % (38/682) of participants with glaucoma knew they had the condition. One in every 5 persons with glaucoma (136;20 %) was blind i.e., visual acuity worse than 3/60. Nigeria has a high prevalence of glaucoma which is largely open-angle glaucoma. A high proportion of those affected are blind. Secondary glaucoma was mostly as a consequence of procedures for cataract. Public health control strategies and high quality glaucoma care service will be required to reduce morbidity and blindness from glaucoma.


1H NMR Spectroscopy Characterization of Porcine Vitreous Humor in Physiological and Photoreceptor Degeneration Conditions.
Qualitative and quantitative analysis of vitreous humor (VH) is important to discriminate between physiological and pathological conditions and may be particularly helpful when using animal models for ophthalmologic research. The aim of the present study was to investigate the physiological qualitative/quantitative composition of the VH of the standard pig using a metabolomics approach based on 1H nuclear magnetic resonance (NMR) spectroscopy. A secondary aim was the characterization of the VH of the porcine model of photoreceptor degeneration induced with iodoacetic acid (IAA), widely used in the biomedical field, and comparison with the physiological one. VH samples were collected from 30 pigs (17 in the physiological condition and 13 treated with IAA) upon vitrectomy and analyzed by means of 1H NMR spectroscopy for characterization. On all analyzed samples, 40 molecules could be identified and quantified, with lactate being the most abundant in both physiological and photoreceptor degeneration conditions. Upon comparison, only 17 molecules showed statistical differences: In the IAA group, glucose and glutamine increased, while lactate, 4-amynobutyrate, hypoxanthine, dicarboxylic acids (succinate and fumarate), amino acids with their derivatives (creatine, alanine, valine, lysine, leucine, glycine, taurine, and isoleucine), and choline with its precursor sn-glycero-3-phosphocholine decreased. The results validate the metabolic impairment determined by glycolysis inhibition upon systemic IAA administration. In conclusion, this work represents the first characterization of the porcine VH metabolome in physiological conditions and provides additional information for the characterization and refinement of the IAA-induced photoreceptor degeneration model.


Histological and transcriptome-wide level characteristics of fetal myofiber hyperplasia during the second half of gestation in Texel and Ujumqin sheep.
Whether myofibers increase with a pulsed-wave mode at particular developmental stages or whether they augment evenly across developmental stages in large mammals is unclear. Additionally, the molecular mechanisms of myostatin in myofiber hyperplasia at the fetal stage in sheep remain unknown. Using the first specialized transcriptome-wide sheep oligo DNA microarray and histological methods, we investigated the gene expression profile and histological characteristics of developing fetal ovine longissimus muscle in Texel sheep (high muscle and low fat), as a myostatin model of natural mutation, and Ujumqin sheep (low muscle and high fat). Fetal skeletal muscles were sampled at 70, 85, 100, 120, and 135 d of gestation. Myofiber number increased sharply with a pulsed-wave mode at certain developmental stages but was not augmented evenly across developmental stages in fetal sheep. The surges in myofiber hyperplasia occurred at 85 and 120 d in Texel sheep, whereas a unique proliferative surge appeared at 100 d in Ujumqin sheep. Analysis of the microarray demonstrated that immune and hematological systems' development and function, lipid metabolism, and cell communication were the biological functions that were most differentially expressed between Texel and Ujumqin sheep during muscle development. Pathways associated with myogenesis and the proliferation of myoblasts, such as calcium signaling, chemokine (C-X-C motif) receptor 4 signaling, and vascular endothelial growth factor signaling, were affected significantly at specific fetal stages, which underpinned fetal myofiber hyperplasia and postnatal muscle hypertrophy. Moreover, we identified some differentially expressed genes between the two breeds that could be potential myostatin targets for further investigation. Proliferation of myofibers proceeded in a pulsed-wave mode at particular fetal stages in the sheep. The myostatin mutation changed the gene expression pattern in skeletal muscle at a transcriptome-wide level, resulting in variation in myofiber phenotype between Texel and Ujumqin sheep during the second half of gestation. Our findings provide a novel and dynamic description of the effect of myostatin on skeletal muscle development, which contributes to understanding the biology of muscle development in large mammals.


Development of the murdoch chiropractic graduate pledge.
This paper reviews the origins of the learned professions, the foundational concepts of professionalism, and the common elements within various healer's oaths. It then reveals the development of the Murdoch Chiropractic Graduate Pledge. A committee comprised of three Murdoch academics performed literature searches on the topic of professionalism and healer's oaths and utilized the Quaker consensus process to develop the Murdoch Chiropractic Graduate Pledge. The committee in its deliberations utilized over 200 relevant papers and textbooks to formulate the Murdoch Chiropractic Graduate Pledge that was administered to the 2010 Murdoch School of Chiropractic and Sports Science graduates. The School of Chiropractic and Sports Science included professionalism as one of its strategic goals and began the process of curriculum review to align it with the goal of providing a curriculum that recognizes and emphasizes the development of professionalism. The reciting of a healer's oath such as the Hippocratic Oath is widely considered to be the first step in a new doctor's career. It is seen as the affirmation that a newly trained health care provider will use his or her newfound knowledge and skill exclusively for the benefit of mankind in an ethical manner. Born from the very meaning of the word profession, the tradition of recitation of a healer's oath is resurgent in health care. It is important for health care instructors to understand that the curriculum must be such that it contributes positively to the students' professional development.


Use of AUDIT-based measures to identify unhealthy alcohol use and alcohol dependence in primary care: a validation study.
As programs for screening, brief intervention, and referral to treatment (SBIRT) for unhealthy alcohol use disseminate, evidence-based approaches for identifying patients with unhealthy alcohol use and alcohol dependence (AD) are needed. While the National Institute on Alcohol Abuse and Alcoholism Clinician Guide suggests use of a single alcohol screening question (SASQ) for screening and Diagnostic and Statistical Manual checklists for assessment, many SBIRT programs use alcohol use disorders identification test (AUDIT) "zones" for screening and assessment. Validation data for these zones are limited. This study used primary care data from a bi-ethnic southern U.S. population to examine the ability of the AUDIT zones and other AUDIT-based approaches to identify unhealthy alcohol use and dependence. Existing data were analyzed from interviews with 625 female and male adult drinkers presenting to 5 southeastern primary care practices. Timeline follow-back was used to identify at-risk drinking, and diagnostic interview schedule was used to identify alcohol abuse and dependence. Validity measures compared performance of AUDIT, AUDIT-C, and AUDIT dependence domains scores, with and without a 30-day binge drinking measure, for detecting unhealthy alcohol use and dependence. Optimal AUDIT scores for detecting unhealthy alcohol use were lower than current commonly used cutoffs (5 for men, 3 for women). Improved performance was obtained by combining AUDIT cutoffs of 6 for men and 4 for women with a 30-day binge drinking measure. AUDIT scores of 15 for men and 13 for women detected AD with 100% specificity but low sensitivity (20 and 18%, respectively). AUDIT dependence subscale scores of 2 or more showed similar specificity (99%) and slightly higher sensitivity (31% for men, 24% for women). Combining lower AUDIT cutoff scores and binge drinking measures may increase the detection of unhealthy alcohol use in primary care. Use of lower cutoff scores and dependence subscale scores may increase diagnosis of AD; however, better measures for detecting dependence are needed.


Dentin protection by a primer-less adhesive technique.
To identify the remaining demineralized dentin (the permeable layer that allows leakage) when using primer-less wet bonding with a total etching period of 10 to 60 seconds compared with contemporary resin adhesives. Extracted human molars were selected and Class V cavities were prepared on the axial surfaces at the CEJ, providing margins in enamel and dentin for leakage testing using a basic dye (n = 10). Seven groups of conditioned teeth [1-1 etched for 10, 30, and 60 seconds (primer-less wet), All-Bond 2 and Single-Bond 2 (wet), AQ-Bond (self-etched), Super-Bond C&B (dry)] were used as bonding systems for resin composite restoration. All specimens were stored in water at 37 degrees C for 24 hours before soaking in a 0.5% basic fuchsin solution for 24 hours. The distance of dye penetration was measured using stereomicroscopy. No leakage at the enamel- and dentin-resin interfaces was found for all etching periods of 1-1 primer-less, Super-Bond C&B, and AQ-Bond specimens. However, leakage was observed at dentin margins in two wet with primer systems, All-Bond 2 (1.31 +/- 1.39 mm) and Single-Bond 2 (0.37 +/- 0.28 mm), which required phosphoric etching to remove a weak smear layer on prepared dentin.


Genome-wide meta-analyses of smoking behaviors in African Americans.
The identification and exploration of genetic loci that influence smoking behaviors have been conducted primarily in populations of the European ancestry. Here we report results of the first genome-wide association study meta-analysis of smoking behavior in African Americans in the Study of Tobacco in Minority Populations Genetics Consortium (n = 32,389). We identified one non-coding single-nucleotide polymorphism (SNP; rs2036527[A]) on chromosome 15q25.1 associated with smoking quantity (cigarettes per day), which exceeded genome-wide significance (β = 0.040, s.e. = 0.007, P = 1.84 × 10(-8)). This variant is present in the 5'-distal enhancer region of the CHRNA5 gene and defines the primary index signal reported in studies of the European ancestry. No other SNP reached genome-wide significance for smoking initiation (SI, ever vs never smoking), age of SI, or smoking cessation (SC, former vs current smoking). Informative associations that approached genome-wide significance included three modestly correlated variants, at 15q25.1 within PSMA4, CHRNA5 and CHRNA3 for smoking quantity, which are associated with a second signal previously reported in studies in European ancestry populations, and a signal represented by three SNPs in the SPOCK2 gene on chr10q22.1. The association at 15q25.1 confirms this region as an important susceptibility locus for smoking quantity in men and women of African ancestry. Larger studies will be needed to validate the suggestive loci that did not reach genome-wide significance and further elucidate the contribution of genetic variation to disparities in cigarette consumption, SC and smoking-attributable disease between African Americans and European Americans.


Fibroblast growth factor 21 deletion aggravates diabetes-induced pathogenic changes in the aorta in type 1 diabetic mice.
Fibroblast growth factor 21 (FGF21) is an important regulator in glucose and lipid metabolism, and has been considered as a potential therapy for diabetes. The effect of FGF21 on the development and progression of diabetes-induced pathogenic changes in the aorta has not currently been addressed. To characterize these effects, type 1 diabetes was induced in both FGF21 knockout (FGF21KO) and C57BL/6 J wild type (WT) mice via multiple-dose streptozotocin injection. FGF21KO diabetic mice showed both earlier and more severe aortic remodeling indicated by aortic thickening, collagen accumulation and fibrotic mediator connective tissue growth factor expression. This was accompanied by significant aortic cell apoptosis than in WT diabetic mice. Further investigation found that FGF21 deletion exacerbated aortic inflammation and oxidative stress reflected by elevated expression of tumor necrosis factor α and transforming growth factor β, and the accumulation of 3-nitrotyrocine and 4-Hydroxynonenal. FGF21 administration can reverse the pathologic changes in FGF21KO diabetic mice. These findings demonstrate that FGF21 deletion aggravates aortic remodeling and cell death probably via exacerbation of aortic inflammation and oxidative stress. This marks FGF21 as a potential therapy for the treatment of aortic damage due to diabetes.


Expression of the neuropeptide Y Y1 receptor in the CNS of rat and of wild-type and Y1 receptor knock-out mice. Focus on immunohistochemical localization.
The distribution of neuropeptide Y (NPY) Y1 receptor-like immunoreactivity (Y1R-LI) has been studied in detail in the CNS of rat using a rabbit polyclonal antibody against the C-terminal 13 amino acids of the rat receptor protein. The indirect immunofluorescence technique with tyramide signal amplification has been employed. For specificity and comparative reasons Y1 knock-out mice and wild-type controls were analyzed. The distribution of Y1R mRNA was also studied using in situ hybridization. A limited comparison between Y1R-LI and NPY-LI was carried out.A widespread and abundant distribution of Y1R-LI, predominantly in processes but also in cell bodies, was observed. In fact, Y1R-LI was found in most regions of the CNS with a similar distribution pattern between rat and wild-type mouse. This staining was specific in the sense that it was absent in adjacent sections following preadsorption of the antibody with 10(-5) M of the antigenic peptide, and that it could not be observed in sections of the Y1 KO mouse. In contrast, the staining obtained with an N-terminally directed Y1R antiserum did not disappear, strongly suggesting unspecificity. In brief, very high levels of Y1R-LI were seen in the islands of Calleja, the anterior olfactory nucleus, the molecular layer of the dentate gyrus, parts of the habenula, the interpeduncular nucleus, the mammillary body, the spinal nucleus of the trigeminal, caudal part, the paratrigeminal nucleus, and superficial layers of the dorsal horn. High levels were found in most cortical areas, many thalamic nuclei, some subnuclei of the amygdaloid complex, the hypothalamus and the nucleus of the stria terminalis, the nucleus of the solitary tract, the parabrachial nucleus, and the inferior olive. Moderate levels of Y1R-LI were detected in the cornu Ammonis and the subicular complex, many septal, some thalamic and many brainstem regions. Y1R staining of processes, often fiber and/or dot-like, and occasional cell bodies was also seen in tracts, such as the lateral lemniscus, the rubrospinal tract and the spinal tract of the trigeminal. There was in general a good overlap between Y1R-LI and NPY-LI, but some exceptions were found. Thus, some areas had NPY innervation but apparently lacked Y1Rs, whereas in other regions Y1R-LI, but no or only few NPY-positive nerve endings could be detected. Our results demonstrate that NPY signalling through the Y1R is common in the rat (and mouse) CNS. Mostly the Y1R is postsynaptic but there are also presynaptic Y1Rs. Mostly there is a good match between NPY-releasing nerve endings and Y1Rs, but 'volume transmission' may be 'needed' in some regions. Finally, the importance of using proper control experiments for immunohistochemical studies on seven-transmembrane receptors is stressed.


Six years' experience with continuous ambulatory peritoneal dialysis and chronic hemodialysis in Saudi children.
Eighty children with End Stage Disease (ESRD) were treated in our unit over a six year period. Forty-eight were treated with CAPD (mean age = 5.8 years) and thirty-two with HD (mean age = 8.2 years). The average duration of treatment was 14.8 months in the CAPD group and 14.2 months in the HD group. There were 22 failures of peritoneal catheter in the CAPD group out of 70 catheters compared to 19 failures of vascular access devices out of 45 in the HD group. Peritoneal catheter failure was due to resistance or recurrent peritonitis in 10 (45.4%) and obstruction in nine (41%), whereas vascular access devices failure was due to thrombosis in six (31.5%) and infection in five (26.3%). Fifteen (31.3%) of the CAPD patients died and eight (16.7%) transferred to HD, whereas five (15.6%) HD patients died and four (12.5%) transferred to CAPD. The three year actuarial rates for CAPD were 81% at one year, 55% at two years and 42% at three years, while for HD was 94%, 85% and 64%, respectively. In this unique experience at the Kingdom so far, we found that a fully integrated service of dialysis including both CAPD and HD are essential. Such a system allows the optimal mode of treatments to be chosen for a child at any time and allows the child to move freely from one treatment to another when needed.


Zinc status in human immunodeficiency virus type 1 infection and illicit drug use.
Zinc deficiency is the most prevalent micronutrient abnormality seen in human immunodeficiency virus (HIV) infection. Low levels of plasma zinc predict a 3-fold increase in HIV-related mortality, whereas normalization has been associated with significantly slower disease progression and a decrease in the rate of opportunistic infections. Studies in Miami, Florida, indicated that HIV-positive users of illicit drugs are at risk for developing zinc deficiency, at least partially because of their poor dietary intake. Zinc deficiency characterized by low plasma zinc levels over time enhances HIV-associated disease progression, and low dietary zinc intake is an independent predictor of mortality in HIV-infected drug users. The amount of zinc supplementation in HIV infection appears to be critical, because deficiency, as well as excessive dietary intake of zinc, has been linked with declining CD4 cell counts and reduced survival. More research is needed to determine the optimal zinc supplementation level in HIV-infected patients, to prevent further burden on an already compromised immune system.


Development of high-density DNA microarray membrane for profiling smoke- and hydrogen peroxide-induced genes in a human bronchial epithelial cell line.
Development of the high-density DNA microarray technique permits the analysis of thousands of genes simultaneously for their differential expression patterns in various biological processes. Through clustering analysis and pattern recognition, the significance of differentially expressed genes can be recognized and correlated with biological events that may take place inside the cell and tissue. With this notion in mind, high-density DNA microarray nylon membrane with colorimetry detection was used to profile the expression of smoke- and hydrogen peroxide-inducible genes in a human bronchial epithelial cell line, HBE1. On the basis of the time course of expression, at least three phases of change in gene expression could be recognized. The first phase is an immediate event in response to oxidant injury. This phase includes induction of the bcl-2 and mdm-2 genes, which are involved in the regulation of apoptosis, and the mitogen-activated protein (MAP) kinase phosphatase 1 (MKP-1) gene, that functions as a regulator of various mitogen-activated protein kinase activities. The second phase, usually 5 h later, includes the induction of various stress proteins and ubiquitin, which are important in providing the chaperone mechanism and the turnover of damaged macromolecules. The third phase, which is 5-10 h later, includes the induction of genes that are apparently involved in reducing oxidative stress by metabolizing reactive oxygen species. In this phase, enzymes associated with tissue and cell remodeling are also elevated. These results demonstrate a complex gene expression array by bronchial epithelial cells in response to the insult of oxidants that are relevant to environmental pollutants.


Bcl-2-related protein A1 is an endogenous and cytokine-stimulated mediator of cytoprotection in hyperoxic acute lung injury.
Hyperoxic acute lung injury (HALI) is characterized by a cell death response with features of apoptosis and necrosis that is inhibited by IL-11 and other interventions. We hypothesized that Bfl-1/A1, an antiapoptotic Bcl-2 protein, is a critical regulator of HALI and a mediator of IL-11-induced cytoprotection. To test this, we characterized the expression of A1 and the oxygen susceptibility of WT and IL-11 Tg(+) mice with normal and null A1 loci. In WT mice, 100% O(2) caused TUNEL(+) cell death, induction and activation of intrinsic and mitochondrial-death pathways, and alveolar protein leak. Bcl-2 and Bcl-xl were also induced as an apparent protective response. A1 was induced in hyperoxia, and in A1-null mice, the toxic effects of hyperoxia were exaggerated, Bcl-2 and Bcl-xl were not induced, and premature death was seen. In contrast, IL-11 stimulated A1, diminished the toxic effects of hyperoxia, stimulated Bcl-2 and Bcl-xl, and enhanced murine survival in 100% O(2). In A1-null mice, IL-11-induced protection, survival advantage, and Bcl-2 and Bcl-xl induction were significantly decreased. VEGF also conferred protection via an A1-dependent mechanism. In vitro hyperoxia also stimulated A1, and A1 overexpression inhibited oxidant-induced epithelial cell apoptosis and necrosis. A1 is an important regulator of oxidant-induced lung injury, apoptosis, necrosis, and Bcl-2 and Bcl-xl gene expression and a critical mediator of IL-11- and VEGF-induced cytoprotection.


[CORRELATION BETWEEN OBESITY AND PANCREATIC CANCER].
Pancreatic cancer is an aggressive tumor with a very poor prognosis, lack of early diagnostic symptoms and highly resistant to therapy. Its incidence is approximately equal to the mortality rate. Even though in recent years progress has been made in defining the morphological and key genetic changes, it is still unclear which factors trigger its occurrence. Some risk factors are age, gender and race, genetic susceptibility, dietary factors, fever, chronic pancreatitis, diabetes and physical inactivity. Studies have shown that an increase in BMI consequently leads to an increased risk of malignancies, including pancreatic cancer. Research based on adipokines and their role in obesity and the occurrence of pancreatic cancer are the potential for a possible future therapeutic interventions.


DEF6, a novel substrate for the Tec kinase ITK, contains a glutamine-rich aggregation-prone region and forms cytoplasmic granules that co-localize with P-bodies.
Localization of DEF6 (SLAT/IBP), a Rho-family guanine nucleotide exchange factor, to the center of the immune synapse is dependent upon ITK, a Tec-family kinase that regulates the spatiotemporal organization of components of T cell signaling pathways and Cdc42-dependent actin polymerization. Here we demonstrate that ITK both interacts with DEF6 and phosphorylates DEF6 at tyrosine residues Tyr(210) and Tyr(222). Expression of a GFP-tagged Y210E-Y222E phosphomimic resulted in the formation of DEF6 cytoplasmic granules that co-localized with decapping enzyme 1 (DCP1), a marker of P-bodies; sites of mRNA degradation. Similarly treatment of cells with puromycin or sodium arsenite, reagents that arrest translation, also resulted in the accumulation of DEF6 in cytoplasmic granules. Bioinformatics analysis identified a glutamine-rich, heptad-repeat region; a feature of aggregating proteins, within the C-terminal region of DEF6 with the potential to promote granule formation through a phosphorylation-dependent unmasking of this region. These data suggest that in addition to its role as a GEF, DEF6 may also function in regulating mRNA translation.


Is body dissatisfaction changing across time? A cross-temporal meta-analysis.
It remains unclear whether body dissatisfaction, a widely recognized predictor of eating-related pathologies and depressive symptomatology, is consistent across cohorts and time. This question is important to investigate because dominant theories propose that sociocultural influences, which may fluctuate, play an important role in the development of body dissatisfaction. Previous efforts for tracking body dissatisfaction across cohorts and time are limited by relying on data from a single institution or using assessments that lack psychometric support across genders. In this study, we utilized cross-temporal meta-analyses to examine changes in 2 dimensions of body dissatisfaction: thinness-oriented dissatisfaction as assessed with the Eating Disorder Inventory-Body Dissatisfaction subscale (data available across 31 years from 326 unique samples, n = 100,228 participants) and muscularity-oriented dissatisfaction as measured with the Drive for Muscularity Scale (data available across 14 years from 117 unique samples, n = 23,575 participants). Results revealed a significant interaction between year of study and gender in predicting thinness-oriented dissatisfaction: girls and women scored higher than boys and men consistently (ds = 0.51-1.17), although only girls' and women's scores decreased gradually across time (d = 0.49). Boys and men scored higher than girls and women on muscularity-oriented dissatisfaction (d = 1.72), with no significant changes across time. These patterns remained when controlling for age and geographic location. Overall, these findings highlight the importance of considering multiple dimensions of body dissatisfaction in research and offer evidence that sociocultural shifts in body acceptance and diversity may be countering thinness-related pressures for girls and women. (PsycINFO Database Record


The Porcine Deltacoronavirus Replication Organelle Comprises Double-Membrane Vesicles and Zippered Endoplasmic Reticulum with Double-Membrane Spherules.
Porcine deltacoronavirus (PDCoV) was first identified in Hong Kong in 2012 from samples taken from pigs in 2009. PDCoV was subsequently identified in the USA in 2014 in pigs with a history of severe diarrhea. The virus has now been detected in pigs in several countries around the world. Following the development of tissue culture adapted strains of PDCoV, it is now possible to address questions regarding virus-host cell interactions for this genera of coronavirus. Here, we presented a detailed study of PDCoV-induced replication organelles. All positive-strand RNA viruses induce the rearrangement of cellular membranes during virus replication to support viral RNA synthesis, forming the replication organelle. Replication organelles for the Alpha-, Beta-, and Gammacoronavirus genera have been characterized. All coronavirus genera induced the formation of double-membrane vesicles (DMVs). In addition, Alpha- and Betacoronaviruses induce the formation of convoluted membranes, while Gammacoronaviruses induce the formation of zippered endoplasmic reticulum (ER) with tethered double-membrane spherules. However, the structures induced by Deltacoronaviruses, particularly the presence of convoluted membranes or double-membrane spherules, are unknown. Initially, the dynamics of PDCoV strain OH-FD22 replication were assessed with the onset of viral RNA synthesis, protein synthesis, and progeny particle release determined. Subsequently, virus-induced membrane rearrangements were identified in infected cells by electron microscopy. As has been observed for all other coronaviruses studied to date, PDCoV replication was found to induce the formation of double-membrane vesicles. Significantly, however, PDCoV replication was also found to induce the formation of regions of zippered endoplasmic reticulum, small associated tethered vesicles, and double-membrane spherules. These structures strongly resemble the replication organelle induced by avian Gammacoronavirus infectious bronchitis virus.


Structure of the native chemotaxis core signaling unit from phage E-protein lysed E. coli cells.
Motile bacteria employ conserved chemotaxis networks to detect chemical gradients in their surroundings and effectively regulate their locomotion, enabling the location of essential nutrients and other important biological niches. The sensory apparatus of the chemotaxis pathway is an array of core-signaling units (CSUs) composed of transmembrane chemoreceptors, the histidine kinase CheA and an adaptor protein, CheW. Although chemotaxis pathways represent the best understood signaling systems, a detailed mechanistic understanding of signal transduction has been hindered by the lack of a complete structural picture of the CSU and extended array. In this study, we present the structure of the complete CSU from phage φX174 E protein lysed Escherichia coli cells, determined using cryo-electron tomography and sub-tomogram averaging to 12-Å resolution. Using AlphaFold2, we further predict the atomic structures of the CSU's constituent proteins as well as key protein-protein interfaces, enabling the assembly an all-atom CSU model, which we conformationally refine using our cryo-electron tomography map. Molecular dynamics simulations of the resulting model provide new insight into the periplasmic organization of the complex, including novel interactions between neighboring receptor ligand-binding domains. Our results further elucidate previously unresolved interactions between individual CheA domains, including an anti-parallel P1 dimer and non-productive binding mode between P1 and P4, enhancing our understanding of the structural mechanisms underlying CheA signaling and regulation. IMPORTANCE Bacterial chemotaxis is a ubiquitous behavior that enables cell movement toward or away from specific chemicals. It serves as an important model for understanding cell sensory signal transduction and motility. Characterization of the molecular mechanisms underlying chemotaxis is of fundamental interest and requires a high-resolution structural picture of the sensing machinery, the chemosensory array. In this study, we combine cryo-electron tomography and molecular simulation to present the complete structure of the core signaling unit, the basic building block of chemosensory arrays, from Escherichia coli. Our results provide new insight into previously poorly-resolved regions of the complex and offer a structural basis for designing new experiments to test mechanistic hypotheses.


Should women aged 70-74 be invited to participate in screening mammography? A report on two Australian community juries.
To elicit informed views from Australian women aged 70-74 regarding the acceptability of ceasing to invite women their age to participate in government-funded mammography screening (BreastScreen). Two community juries held in 2017. Greater Sydney, a metropolis of 4.5 million people in New South Wales, Australia. 34 women aged 70-74 with no personal history of breast cancer, recruited by random digit dialling and previously randomly recruited list-based samples. Jury verdict and rationale in response to structured questions. We transcribed audio-recorded jury proceedings and identified central reasons for the jury's decision. The women's average age was 71.5 years. Participants were of diverse sociocultural backgrounds, with the sample designed to include women of lower levels of educational attainment. Both juries concluded by majority verdict (16-2 and 10-6) that BreastScreen should continue to send invitations and promote screening to their age group. Reasons given for the majority position include: (1) sending the invitations shows that society still cares about older women, empowers them to access preventive health services and recognises increasing and varied life expectancy; (2) screening provides women with information that enables choice and (3) if experts cannot agree, the conservative approach is to maintain the status quo until the evidence is clear. Reasons for the minority position were the potential for harms through overdiagnosis and misallocation of scarce health resources. Preventive programmes such as mammography screening are likely to have significant symbolic value once they are socially embedded. Arguments for programme de-implementation emphasising declining benefit because of limited life expectancy and the risks of overdiagnosis seem unlikely to resonate with healthy older women. In situations where there is no consensus among experts on the value of established screening programmes, people may strongly prefer receiving information about their health and having the opportunity make their own choices.


Genome-wide analysis of the Drosophila immune response by using oligonucleotide microarrays.
To identify new Drosophila genes involved in the immune response, we monitored the gene expression profile of adult flies in response to microbial infection by using high-density oligonucleotide microarrays encompassing nearly the full Drosophila genome. Of 13,197 genes tested, we have characterized 230 induced and 170 repressed by microbial infection, most of which had not previously been associated with the immune response. Many of these genes can be assigned to specific aspects of the immune response, including recognition, phagocytosis, coagulation, melanization, activation of NF-kappaB transcription factors, synthesis of antimicrobial peptides, production of reactive oxygen species, and regulation of iron metabolism. Additionally, we found a large number of genes with unknown function that may be involved in control and execution of the immune response. Determining the function of these genes represents an important challenge for improving our knowledge of innate immunity. Complete results may be found at http://www.fruitfly.org/expression/immunity/.


HIV-1 tat protein induces a migratory phenotype in human fetal microglia by a CCL2 (MCP-1)-dependent mechanism: possible role in NeuroAIDS.
Acquired immune deficiency syndrome (AIDS) encephalitis and dementia are characterized by neuronal loss, astrogliosis, and microglia activation and migration that contribute to the formation of multinucleated giant cells. Despite extensive evidence of pathological changes in the brain of infected individuals, the mechanisms of human immune deficiency virus type 1 (HIV-1) entry, microglia migration, and viral propagation within the brain are still not completely understood. In this study, we report that the induction of a migratory phenotype in human fetal microglia by the HIV-1 transactivator protein, tat, is mediated by the chemokine, CCL2. CCL2 or tat treatment alone induced rearrangement of actin and the formation of microglial processes. The time course of cell membrane ruffling induced by CCL2 was faster (5-30 min) than that elicited by tat treatment (2-3 h). Our previous data in human fetal microglia showed that tat induces CCL2 expression. Thus, we examined whether tat-induced microglia membrane ruffling and process formation, critical components in cell migration, are mediated by the secretion of CCL2 by these cells. To test this hypothesis, we treated microglia with tat protein in the presence of neutralizing CCL2 antibodies. Co-treatment with neutralizing CCL2 antibodies resulted in the loss of tat-induced membrane ruffling. Tat treatment of microglia induced polarization of CCR2, the receptor for CCL2, to the leading edge of processes, further suggesting a CCL2-dependent mechanism of tat-induced microglia migration. Our data indicate that tat facilitates microglia migration by inducing autocrine CCL2 release. Our results suggest that tat induced CCL2 secretion may be one of the early signals during NeuroAIDS.


Doctor-patient interaction for irritable bowel syndrome in primary care: a systematic perspective.
Irritable bowel syndrome (IBS) is defined by specific validated symptom criteria and encompasses several different underlying pathophysiological mechanisms that express a common set of symptoms. However, IBS is poorly understood by patients. We aimed to explore how a diagnosis of IBS affects the interaction between patients and their physicians. A comprehensive literature search for studies in the English language addressing this issue was conducted using Medline, PubMed, Cochrane Database, Psychinfo, Cinahl, Embase, Web of Science and manual recursive search of reference lists. Investigators reviewed and abstracted data from articles fulfilling our inclusion criteria: primary care patients, all ages, gender and ethnic groups diagnosed with IBS by a general practitioner (GP). Retrieval of 121 articles generated only four that met inclusion criteria. Research methods of three studies relied solely on qualitative subjective, anecdotal patient narratives, a bias in favor of patients' negative opinion, absence of objective physician diagnostic criteria, pre-testing questions for two studies, follow-up and patient verification of accounts for accuracy. The fourth study included objective physician diagnostic criteria, quantitative measures, a pre-testing questionnaire, and both patient and doctor perspectives. There was a disparity between patient and GP perception regarding the nature, severity and consequences of IBS in primary care, leading patients to perceive this interaction as one of dissatisfaction. The fourth study revealed GP management of IBS mostly meets patient's expectations areas of concern centered on etiology, diagnostic criteria and dietary advice. Disparity seems to lie with the physician, who needs to provide more trust, knowledge, and sympathy, create rapport and be forthcoming with information, while keeping information simple and understandable. Patient dissatisfaction stems from the actual information provided and how this is communicated. There is evidence that some IBS patients in primary care experience dissatisfaction and negative attitudes in GP interactions. Future research should take into account personality attributes and cross-situational stability in addition to methodological implications of studies. GPs may be the first avenue for IBS patients to vent their frustration, and appropriate education programs for optimal management of patients with IBS are needed in primary care.


Colistin-resistant Acinetobacter baumannii: beyond carbapenem resistance.
With an increase in the use of colistin methansulfonate (CMS) to treat carbapenem-resistant Acinetobacter baumannii infections, colistin resistance is emerging. Patients with infection or colonization due to colistin-resistant A. baumannii were identified at a hospital system in Pennsylvania. Clinical data were collected from electronic medical records. Susceptibility testing, pulsed-field gel electrophoresis (PFGE), and multilocus sequence typing (MLST) were performed. To investigate the mechanism of colistin resistance, lipid A was subjected to matrix-assisted laser desorption/ionization mass spectrometry. Twenty patients with colistin-resistant A. baumannii were identified. Ventilator-associated pneumonia was the most common type of infection. Nineteen patients had received intravenous and/or inhaled CMS for treatment of carbapenem-resistant, colistin-susceptible A. baumannii infection prior to identification of colistin-resistant isolates. The 30-day all-cause mortality rate was 30%. The treatment regimen for colistin-resistant A. baumannii infection associated with the lowest mortality rate was a combination of CMS, a carbapenem, and ampicillin-sulbactam. The colistin-susceptible and -resistant isolates from the same patients were highly related by PFGE, but isolates from different patients were not, suggesting evolution of resistance during CMS therapy. By MLST, all isolates belonged to the international clone II, the lineage that is epidemic worldwide. Phosphoethanolamine modification of lipid A was present in all colistin-resistant A. baumannii isolates. Colistin-resistant A. baumannii occurred almost exclusively among patients who had received CMS for treatment of carbapenem-resistant, colistin-susceptible A. baumannii infection. Lipid A modification by the addition of phosphoethanolamine accounted for colistin resistance. Susceptibility testing for colistin should be considered for A. baumannii identified from CMS-experienced patients.


Role of stereotactic biopsy in multifocal brain lesions: considerations on 100 consecutive cases.
One hundred patients affected by multifocal brain lesions were investigated by serial stereotactic biopsy. Systemic diseases and primary neoplasms elsewhere were previously ruled out. The histological diagnosis obtained in this series comprises malignant gliomas in 37% of patients; primary non-Hodgkin's brain lymphoma in 15%; metastatic brain tumours in 15% (no evidence of the primary tumour at the time of stereotactic surgery); low grade gliomas in 12%; infective diseases in 10% (including brain abscesses and multifocal viral encephalitis); and ischaemic lesions in 6%. In addition, two patients with germinomas, two with primitive neuroepithelial tumours, two with multiple telangiectases, and one with a teratoma were also included in this series. Histological findings obtained by stereotactic procedures guided the choice of treatment, avoiding the risks of blind treatments. Indications and future perspectives for stereotactic surgery in multifocal brain lesions are discussed with emphasis on advances in diagnostic and therapeutic tools.