Genes from two intermoult puffs in Drosophila virilis polytene chromosomes are differentially transcribed during larval development.
Genes from two Drosophila virilis intermoult puffs were isolated by microcloning. From puff 16A on the X-chromosome a 2.9 kb DNA fragment was obtained, which hybridizes with three transcripts. Two of them represent the mRNAs for larval glue proteins. They are found in different abundancies in third larval instar salivary glands, but also in minor amounts in midgut and in fat body. In puff 55E on chromosome III two genes were identified. They are transcribed exclusively in salivary glands during all three larval instars. Therefore, their products must be related to another gland-specific function, which is sustained throughout larval life.


Transurethral resection of fibrotic scar tissue combined with temporary urethral stent placement for patients with in anterior urethral stricture.
Fibrotic scar formation is a main cause of recurrent urethral stricture after initial management with direct vision internal urethrotomy (DVIU). In the present study, we devised a new technique of combined the transurethral resection of fibrotic scar tissue and temporary urethral stenting, using a thermo-expandable urethral stent (Memokath(TM) 044TW) in patients with anterior urethral stricture. As a first step, multiple incisions were made around stricture site with cold-utting knife and Collins knife electrode to release a stricture band. Fibrotic tissue was then resected with a 13Fr pediatric resectoscope before deployment of a MemokathTM 044TW stent (40 - 60mm) on a pre-mounted sheath using 0° cystoscopy. Stents were removed within12 months after initial placement. We performed this technique on 11 consecutive patients with initial (n = 4) and recurrent (n = 7) anterior urethral stricture (April 2009 – February 2013). At 18.9 months of mean follow-up (12-34 months), mean Qmax (7.8±3.9ml/sec vs 16.8 ± 4.8ml/sec, p < 0.001), IPSS (20.7 vs 12.5, p = 0.001 ), and QoL score (4.7 vs 2.2, p < 0.001) were significantly improved. There were no significant procedure-related complications except two cases of tissue ingrowth at the edge of stent, which were amenable by transurethral resection. In 7 patients, an average 1.4 times (1-5 times) of palliative urethral dilatation was carried out and no patients underwent open surgical urethroplasty during the follow-up period. Combined transurethral resection and temporary urethral stenting is a effective therapeutic option for anterior urethral stricture. Further investigations to determine the long-term effects, and safety profile of this new technique are warranted.


Cost-effectiveness of UGT1A1*28 genotyping in preventing severe neutropenia following FOLFIRI therapy in colorectal cancer.
Functional polymorphisms of the UGT1A1 gene, particularly the UGT1A1*28 variant, are associated with the severity of the bone marrow suppression in patients with metastatic colorectal cancer receiving irinotecan. This study assesses the cost-effectiveness of screening for UGT1A1*28 polymorphism associated with primary prophylactic Granulocytes Colony Stimulating Factor in patients homozygous for the *28 allele. The effectiveness was estimated based on the number of neutropenia avoided. We modelled a theoretical population treated with combined 5-fluorouracil, leucovorin and irinotecan (FOLFIRI) for metastatic colorectal cancer. A decision tree simulated the health outcomes, measured by the prevalence of neutropenic events for two strategies, with or without UGT1A1 genotype screening. The model incorporated direct hospital costs and was validated with a sensitivity analysis. We calculated the cost-effectiveness ratio: CE=∆C / ∆E = "genotyping" cost - "no genotyping" cost / number of febrile neutropenia avoided. In the "genotyping strategy", the cost to avoid one febrile neutropenia event per 1000 patients treated was € 942.8 to € 1090.1. The sensitivity analysis showed a better CE ratio of € 733.4 to € 726.6 per febrile neutropenic event avoided. UGT1A1 genotype screening before irinotecan treatment is a cost-efficient strategy for the hospital. Systematic genotyping prior to chemotherapy, and administration of CSF in patients homozygotes for the *28 allele allow to avoid 91 febrile neutropenias at an acceptable cost.


Minimally invasive surgery for gastric cancer.
The interest in minimally invasive surgery (MIS) has rapidly increased in recent decades and surgeons have adopted minimally invasive techniques due to its reduced invasiveness and numerous advantages for patients. With increased surgical experience and newly developed surgical instruments, MIS has become the preferred approach not only for benign disease but also for oncologic surgery. Recently, robotic systems have been developed to overcome difficulties of standard laparoscopic instruments during complex procedures. Its advantages including three-dimensional images, tremor filtering, motion scaling, articulated instruments, and stable retraction have created the opportunity to use robotic technology in many procedures including cancer surgery. Gastric cancer is one of the most common causes of cancer-related deaths worldwide. While its overall incidence has decreased worldwide, the proportion of early gastric cancer has increased mainly in eastern countries following mass screening programs. The shift in the paradigm of gastric cancer treatment is toward less invasive approaches in order to improve the patient's quality of life while adhering to oncological principles. In this review, we aimed to summarize the operative strategy and current literature in laparoscopic and robotic surgery for gastric cancer.


The roles of two transcription factors, ABI4 and CBFA, in ABA and plastid signalling and stress responses.
Genetic and physiological studies have revealed evidences for multiple signaling pathways by which the plastid exerts retrograde control over photosynthesis-associated-nuclear-genes. In this study we have examined the mechanisms of control of transcription by plastid signals, focusing on transcription factors. We have also further addressed the physical nature of plastid signals and the physiological role, in stress acclimation of this regulatory pathway. ABI4, a master Apetala 2 (AP2)-type transcription factor (TF), is targeted by multiple signalling pathways in plant cells, such as abscisic acid (ABA) signals, sugar signals and plastid signals derived from reactive oxygen species (ROS) and chlorophyll intermediates. ABI4 binds the promoter of target genes to prevent their transcription by competing with other competitive TFs. However, we found that once ABI4 bound the element (CCACGT), it may not be bound by other TFs, therefore making the signalling long-lasting. Downstream of ABI4, CBFA (CCAAT binding factor A) is a subunit of the HAP2/HAP3/HAP5 (Heme activator protein) trimeric transcription complex. CBFA however is a redundant HAP3 subunit. When emergency occurs (such as herbicide treatments or environmental stresses followed by ABA and ROS accumulation), the master transcription factor ABI4 down-regulates some TFs, like CBFA, and then some other TF subunits enter the transcription complex and transcriptional efficiency of stress-responsive genes (including the transcription co-factor CBP) is improved instantaneously. abi4, cbfA and cbp mutants showed weaker drought-tolerance after a herbicide norflurazon treatment, which indicated the physiological role of these key transcription factors.


Endogenous gut-derived bacterial endotoxin tonically primes pancreatic secretion of insulin in normal rats.
This laboratory has proposed that endogenous gut-derived bacterial endotoxin primes the pancreatic secretion of insulin in normal rats. Endogenous lipopolysaccharide (LPS) is continually absorbed from the gut into intestinal capillaries, and low-grade portal venous endotoxemia is the status quo. Under physiologic conditions, Kupffer cells of the liver totally phagocytize and degrade endotoxin from the portal circulation. Evidence from this and other laboratories indicates that administration of exogenous LPS to humans and rats enhances pancreatic secretion of both insulin and glucagon. Conversely, findings of the present study demonstrate that restriction of endogenous LPS in fasted rats depresses the basal and arginine-stimulated concentrations of plasma insulin. Techniques used to restrict gut-derived LPS availability included chronic daily gavage with neomycin and cefazolin for gut sterilization and with cholestyramine or lactulose to reduce endotoxin within the gut. In addition, induction of endotoxin tolerance was produced by progressively higher doses of LPS intraperitoneally (i.p.), and polymyxin B was administered subcutaneously (s.c.) daily to neutralize the lipid A portion of circulating LPS. Finally, isolator-reared, defined flora rats, which were gram-negative-bacteria-deficient, and, therefore, LPS-deficient, were compared with conventional counterparts. Basal plasma insulin but not glucagon levels were consistently and significantly reduced in endogenous LPS-restricted animals. Glucose-stimulated plasma insulin was decreased only after parenteral treatment by tolerance induction and polymyxin B administration. Both plasma insulin and glucagon were depressed in response to arginine challenge in most LPS-restricted rats.(ABSTRACT TRUNCATED AT 250 WORDS)


Indoor and outdoor monitoring of volatile organic compounds in school buildings: indicators based on health risk assessment to single out critical issues.
Children are more sensitive to pollutants than adults and yet they spend large amounts of time in school environments where they are exposed to unknown levels of indoor pollutants. This study investigated the concentrations of the most abundant volatile organic compounds (VOCs) in eight naturally ventilated school buildings in Italy. The schools were chosen to include areas with different urbanization and traffic density characteristics in order to gather a more diverse picture of exposure risks in the different areas of the city. VOCs were sampled for one week in the presence/absence of pupils using diffusive samplers suitable for thermal desorption inside three classrooms at each school. The samples were then analyzed with thermal desorption-gas chromatography-mass spectrometry (TD-GC-MS). In addition, outdoor measurements were carried out in the yard at each school. VOC identification and quantification, and indoor/outdoor concentration plots were used to identify pollutant sources. While some classrooms were found to have very low VOC levels, others had a significant indoor contribution or a prevalent outdoor contribution. High concentrations of terpenes were found in all monitored classrooms: a-pinene and limonene were in the range of 6.55-34.18 µg/m3 and 11.11-25.42 µg/m3 respectively. Outdoor concentrations were lower than indoors for each monitored school. Indicators based on health risk assessment for chronic health effects associated with VOCs (either carcinogenic or non-carcinogenic) were proposed to rank sites according to their hazard level.


A developmental staging series for the lizard genus Anolis: a new system for the integration of evolution, development, and ecology.
Vertebrate developmental biologists typically rely on a limited number of model organisms to understand the evolutionary bases of morphological change. Unfortunately, a typical model system for squamates (lizards and snakes) has not yet been developed leaving many fundamental questions about morphological evolution unaddressed. New model systems would ideally include clades, rather than single species, that are amenable to both laboratory studies of development and field-based analyses of ecology and evolution. Combining an understanding of development with an understanding of ecology and evolution within and between closely related species has the potential to create a seamless understanding of how genetic variation underlies ecologically and evolutionarily relevant variation within populations and between species. Here we briefly introduce a new model system for the integration of development, evolution, and ecology, the lizard genus Anolis, a diverse group of lizards whose ecology and evolution is well understood, and whose genome has recently been sequenced. We present a developmental staging series for Anolis lizards that can act as a baseline for later comparative and experimental studies within this genus.


The influence of training experiences on career intentions of the future GP workforce: a qualitative study of new GPs in England.
The capacity of the UK GP workforce has not kept pace with increasing primary care workloads. Although many doctors successfully complete GP specialty training programmes, some do not progress to work in NHS general practice. This article explores the training experiences and perceptions of newly qualified GPs to understand how their education, training, and early experiences of work influence their career plans. A qualitative study of doctors in their final year of GP training (ST3) and within 5 years of completion of GP training (F5). Participants across England were recruited through training programmes, First5 groups, and publicity using social media and networks. Open narrative interviews were conducted with individuals and focus groups. Audiorecorded interviews were transcribed, and a thematic analysis was supported by NVivo and situational analysis mapping techniques. Fifteen participants engaged in individual interviews and 10 focus groups were carried out with a total of 63 participants. Most doctors reported that training programmes had prepared them to deal confidently with most aspects of routine clinical GP work. However, they felt underprepared for the additional roles of running a practice and in their understanding of wider NHS organisational structures. Doctors wished to avoid unacceptably heavy workloads and voiced concerns about the longer-term sustainability of general practice. Strategies to attract and retain enough GPs to support delivery of comprehensive primary care should consider how doctors' early career experiences influence their career intentions. A coherent plan is needed to improve their preparation and increase confidence that they can achieve a professionally satisfying, effective, and sustainable career in NHS general practice.


Arsenic in private well water part 1 of 3: Impact of the New Jersey Private Well Testing Act on household testing and mitigation behavior.
Regularly ingesting water with elevated arsenic increases adverse health risks. Since September 2002, the NJ Private Well Testing Act (PWTA) has required testing untreated well water for arsenic during real estate transactions in 12 counties. Its implementation provides an opportunity to investigate the effects of policy intervention on well testing and treatment behavior. Here we analyze results of a survey mailed to 1943 random addresses (37% response), including responses from 502 private well households who purchased their homes prior to PWTA commencement and 168 who purchased after. We find the PWTA has significantly increased arsenic testing rates in an area where 21% of wells contain arsenic above the 5μg/L NJ drinking water standard. The PWTA has allowed identification of more wells with arsenic (20% of post-PWTA vs. 4% of pre-PWTA households) and more treatment for arsenic (19% of post-PWTA vs. 3% of pre-PWTA households). Such an Act is a partial answer to significant socioeconomic disparities in testing observed among households for whom it is not required. Additionally residents purchasing homes since 2002 are younger and disproportionately more likely to have children in their household (60% vs. 32%), a priority group given their particular vulnerability to effects of arsenic. Despite more wells tested under the PWTA, post-PWTA well owners forget or misremember arsenic test results more often, are more likely to report not knowing what kind of treatment they are using, and are not reporting better maintenance or monitoring of their treatment systems than pre-PWTA households. This suggests serious challenges to reducing arsenic exposure remain even when testing is a requirement. Furthermore, only a fraction of wells have been tested under the PWTA due to the slow pace of housing turnover. We recommend more public resources be made available to support private well testing among socially and biologically vulnerable groups.


Dynamic wettability of polyethylene glycol-modified poly(dimethylsiloxane) surfaces in an aqueous/organic two-phase system.
We herein report the preparation of a surface that behaves in a hydrophobic manner but does not undergo protein adsorption in an aqueous/organic two-phase system. We found that polyethylene-glycol (PEG)-modified poly(dimethylsiloxane) (PDMS) exhibits hydrophobic properties when the surface is immersed in an organic solution, while the PEG moiety prevents protein adsorption on the PDMS surface in an aqueous solution at high protein concentrations due to the dynamic behaviour of the PEG moiety. As such, we demonstrated the in-well droplet formation of an aqueous solution containing a high protein concentration. In addition, to demonstrate the feasibility of this method in single cell analyses, a droplet array of a liquid medium containing 10% fetal bovine serum and HeLa cells was formed. The preparation of a droplet array using our PDMS-PEG surface to promote in-well droplet formation avoided the use of flow control equipment and complicated microstructures. We therefore expect that the dynamic wettability of our reported surface will be applicable in single cell and biochemical analyses, such as protein characterisation using crystallography or immunoassays.


Validation of fatty acid intakes estimated by a food frequency questionnaire using erythrocyte fatty acid profiling in the Montreal Heart Institute Biobank.
To improve the prevention, treatment and risk prediction of cardiovascular diseases, genetic markers and gene-diet interactions are currently being investigated. The Montreal Heart Institute (MHI) Biobank is suitable for such studies because of its large sample size (currently, n = 17 000), the availability of biospecimens, and the collection of data on dietary intakes of saturated (SFAs) and n-3 and n-6 polyunsaturated (PUFAs) fatty acids estimated from a 14-item food frequency questionnaire (FFQ). We tested the validity of the FFQ by correlating dietary intakes of these fatty acids with their red blood cell (RBC) content in MHI Biobank participants. Seventy-five men and 75 women were selected from the Biobank. We successfully obtained RBC fatty acids for 142 subjects using gas chromatography coupled to mass spectrometry. Spearman correlation coefficients were used to test whether SFA scores and daily intakes (g day(-1)) of n-3 and n-6 PUFAs correlate with their RBC content. Based on covariate-adjusted analyses, intakes of n-3 PUFAs from vegetable sources were significantly correlated with RBC α-linolenic acid levels (ρ = 0.23, P = 0.007), whereas n-3 PUFA intakes from marine sources correlated significantly with RBC eicosapentaenoic acid (ρ = 0.29, P = 0.0008) and docosahexaenoic acid (ρ = 0.41, P = 9.2 × 10(-7)) levels. Intakes of n-6 PUFAs from vegetable sources correlated with RBC linoleic acid (ρ = 0.18, P = 0.04). SFA scores were not correlated with RBC total SFAs. The MHI Biobank 14-item FFQ can appropriately estimate daily intakes of n-3 PUFAs from vegetable and marine sources, as well as vegetable n-6 PUFAs, which enables the possibility of using these data in future studies.


Noninvasive and prospective diagnosis of coronary heart disease with urine using surface-enhanced Raman spectroscopy.
A prospective diagnosis method for coronary heart disease (CHD) using human urine based on surface-enhanced Raman spectroscopy (SERS) is proposed, and could provide valuable information for judging whether to perform percutaneous coronary intervention (PCI) in clinics. Here, urine samples from 87 patients with CHD, including patients with PCI before operation (degree of cardiovascular congestion above 70%) and without PCI (degree of cardiovascular congestion under 70%), and 20 healthy humans were measured using SERS. Principal component analysis (PCA) combined with linear discriminant analysis (LDA) was employed to analyze the SERS spectra, revealing that the classification sensitivity and specificity were 90% and 78.9%, respectively, and the absolute value for loading of PC1 at 1509 cm-1 was the largest. Since platelet-derived growth factor-BB (PDGF-BB) is closely related to CHD, PDGF-BB aqueous solutions with various concentrations (1, 0.5, 0.1, 0.05 and 0.01 ppm) and a mixture of healthy human urine and PDGF-BB aqueous solutions were then investigated in this work, and it was found that the Raman peak at 1509 cm-1 may be attributed to PDGF-BB. Moreover, the measured SERS spectra of all the urine samples from the 87 patients with CHD were compared with the clinical data provided by a hospital, and it was revealed that the appearance of a peak at 1509 cm-1 in the SERS spectra was in good agreement with the results of coronary angiography tests when cardiovascular congestion was above 70%. This indicated that the classification sensitivity and specificity were 87.9% and 87.0%, respectively, through identification of the Raman peak at 1509 cm-1.


Are Patient Self-Reported Outcome Measures Sensitive Enough to Be Used as End Points in Clinical Trials?: Evidence from the United Kingdom Glaucoma Treatment Study.
The United Kingdom Glaucoma Treatment Study (UKGTS) demonstrated the effectiveness of an intraocular pressure-lowering drug in patients with glaucoma using visual field progression as a primary outcome. The present study tested the hypothesis that responses on patient-reported outcome measures (PROMs; secondary outcome measure) differ between patients receiving a topical prostaglandin analog (latanoprost) or placebo eye drops in UKGTS. Multicenter, randomized, triple-masked, placebo-controlled trial. Newly diagnosed glaucoma patients in the UKGTS with baseline and exit PROMs (n = 182 and n = 168 patients from the treatment and placebo groups, respectively). In the UKGTS (trial registration number, ISRCTN96423140), patients with open-angle glaucoma were allocated to receive latanoprost (treatment) or placebo; the observation period was 24 months. Patients completed general health PROMs (European Quality of Life in 5 Dimensions [EQ-5D] and 36-item Short Form [SF-36]) and PROMs specific to glaucoma (15-item Glaucoma Quality of Life [GQL-15] and 9-item Glaucoma Activity Limitation [GAL-9]) at baseline and exit from the trial. Percentage changes between measurement on PROMs were calculated for each patient and compared between treatment arms. In addition, differences between stable patients (n = 272) and those with glaucomatous progression (n = 78), as determined by visual field change (primary outcome), were assessed. PROMs on health-related and vision-related quality of life. Average percentage change on PROMs was similar for patients in both arms of the trial, with no statistically significant differences between treatment and placebo groups (EQ-5D, P = 0.98; EQ-5D visual analog scale, P = 0.88; SF-36, P = 0.94, GQL-15, P = 0.66; GAL-9, P = 0.87). There were statistically significant differences between stable and progressing patients on glaucoma-specific PROMs (GQL-15, P = 0.02; GAL-9, P = 0.02), but not on general health PROMs (EQ-5D, P = 0.62; EQ-5D visual analog scale, P = 0.23; SF-36, P = 0.65). Average change in PROMs on health-related and vision-related quality of life was similar for the treatment and placebo groups in the UKGTS. The PROMs used may not be sensitive enough to function as primary end points in clinical trials when participants have newly diagnosed early-stage glaucoma.


The expansion of amino-acid repeats is not associated to adaptive evolution in mammalian genes.
The expansion of amino acid repeats is determined by a high mutation rate and can be increased or limited by selection. It has been suggested that recent expansions could be associated with the potential of adaptation to new environments. In this work, we quantify the strength of this association, as well as the contribution of potential confounding factors. Mammalian positively selected genes have accumulated more recent amino acid repeats than other mammalian genes. However, we found little support for an accelerated evolutionary rate as the main driver for the expansion of amino acid repeats. The most significant predictors of amino acid repeats are gene function and GC content. There is no correlation with expression level. Our analyses show that amino acid repeat expansions are causally independent from protein adaptive evolution in mammalian genomes. Relaxed purifying selection or positive selection do not associate with more or more recent amino acid repeats. Their occurrence is slightly favoured by the sequence context but mainly determined by the molecular function of the gene.


lncRNA-Encoded Polypeptide SPAR(s) with mTORC1 to Regulate Skeletal Muscle Regeneration.
Although prematurely baptized as non-coding, some lncRNAs encode polypeptides with regulatory functions that are implicated in various biological processes. Matsumoto et al. (2017) recently report in Nature that LINC00961 generates SPAR polypeptide that acts via the lysosome to suppress amino-acid-mediated mTORC1 activity, thereby modulating skeletal muscle regenerative response following injury.


The effect of the Müller-Lyer illusion on the planning and control of manual aiming movements.
Two experiments used Müller-Lyer stimuli to test the predictions of the planning-control model (S. Glover, 2002) for aiming movements. In Experiment 1, participants aimed to stimuli that either remained the same or changed upon movement initiation. Experiment 2 was identical except that the duration of visual feedback for online control was manipulated. The authors found that the figures visible during movement planning and online control had additive effects on endpoint bias, even when participants had ample time to use visual feedback to modify their movements (Experiment 2). These findings are problematic not only for the planning-control model but also for A. D. Milner and M. A. Goodale's (1995) two visual system explanation of illusory bias. Although our results are consistent with the idea that a single representation is used for perception, movement planning, and online control (e.g., V. H. Franz, 2001), other work from our laboratory and elsewhere suggests that the manner in which space is coded depends on constraints associated with the specific task, such as the visual cues available to the performer.


The effect of calcium on lead absorption in rats.
The effects of Ca2+ on lead absorption as PbCl2 and 203PbCl2 were studied in rats. 1. Doubling of dietary calcium with Ca3(PO4)2 significantly decreased lead absorption as assessed by 203Pb retention, tissue lead concentration, urinary excretion of delta-aminolaevulinate and increased activities of delta-aminolaevulinate dehydratase and ferrochelatase. 2. Similar effects on lead absorption were shown by the Ca2+ salts, Cl-, CO32-, PO43-, SO42-, gluconate and glycerophosphate. 3. CaCl2 and calcium glycerophosphate were found to be most effective in decreasing lead absorption when administered immediately before lead dosage. 4. A negative exponential relationship was found between CaCl2 concentration and 203Pb absorption at 120h. The result suggest that, above 4 mmol of administered calcium, residual lead absorption is unaffected by increasing gastrointestinal calcium concentrations. 5. Increased systemic calcium had no effect on lead retention. 6. Calcium in the concentrations found in domestic hard-water supplies significantly decreased absorption of a solution of 203Pb dissolved in it compared with absorption of 203Pb dissolved in soft or distilled water. 7. Milk and skimmed milk were found to have no effect on 203Pb absorption in rats.


Three patients resembling Teebi-Shaltout syndrome.
Teebi-Shaltout syndrome (TSS) was first reported by Teebi and Shaltout in 1989. This entity is proposed to be inherited in autosomal recessive manner. The clinical features include characteristic facial features, ectodermal dysplasia, camptodactyly, and caudal appendage. Only one additional paper reporting four additional cases has been published since the first description. Clinical features common to all previously affected individuals diagnosed with TSS are craniofacial, orodental-ectodermal, and skeletal. This report summarizes and discusses the findings of three additional patients from two unrelated families with findings similar to TSS. These findings may be present in a genetically and phenotypically heterogeneous group of disorders similar to TSS. Presence of consanguinity and similarly affected siblings of both genders suggests autosomal recessive inheritance.


Rapid diagnosis of micrometastasis of gastric cancer using reverse transcription loop-mediated isothermal amplification.
Methods to detect metastases in biopsy specimens with certain rapidity and accuracy are essential to performing tailor-made surgeries for solid malignancies. Reverse transcription loop-mediated isothermal amplification (RT-LAMP) reaction is a novel technique for detecting mRNA expression of target sequences with high sensitivity and rapidity, even from crude samples without RNA purification. Applicability to detect lymph node (LN) micrometastasis of gastric cancer was tested. Total of 26 LNs were retrieved from 10 patients with primary gastric cancer. Each LN was serially sectioned, and every set of three serial sections were tested for routine histopathological (H&E) and immunohistochemical examination with anti-cytokeratin antibodies (IHC), and RT-LAMP analysis targeted cytokeratin 19 mRNA. Results from H&E/IHC and RT-LAMP analysis were compared in each set of sections. All the sections of those containing metastatic lesions equivalent to a volume of overt metastasis (maximum diameter>2 mm), 90% of those containing micrometastasis (between 2 and 0.2 mm) and 83% of those containing isolated tumor cells (<0.2 mm) were detectable using this procedure. Total analysis from lysates of clinical specimens required<75 min. This new technique is suggested to be an alternative to rapid diagnosis of micrometastasis based on conventional histopathological analysis.


Fluidic actuation of an elastomeric grating.
A fluidic chamber with an elastomeric grating membrane is fabricated. Grating groove spacing is modified through membrane deformation via fluid injection. Tunable diffraction output is demonstrated. At normal incidence, the diffraction angle changes by 14.2° and 9.8° for incident wavelengths 632.8 and 488 nm, respectively, with an injected fluid volume of 1 ml.


Prevalence of food security in the UK measured by the Food Insecurity Experience Scale.
Measurement of UK food insecurity has historically been inconsistent, making it difficult to understand trends. This study contributes by reporting and analysing data from a national survey conducted in line with UN Food and Agriculture Organization (FAO) recommendations and standard methods, providing an internationally comparable pre-coronavirus disease 2019 (COVID-19) snapshot of food insecurity. Data came from a nationally representative 2019 UK sample (N = 2000) surveyed by Ipsos-Mori. Prevalence of food insecurity was assessed using the UN FAO Food Insecurity Experience Scale. Logistic regression was used to model food insecurity in relation to geographic and socio-demographic variables. Severe food insecurity was reported by 3% of the sample, an increase of 66.7% over the last directly comparable UK analysis (Gallup World Poll data from 2016 to 2018). Indication of some degree of food insecurity was reported by 14.2% of the sample and tended to be higher amongst younger age groups, those on lower incomes, and home renters (as opposed to owners). No geographic variables were significantly associated with food insecurity prevalence. The finding that prevalence of severe food insecurity was already increasing before the COVID-19 pandemic, across all areas of the UK, is cause for concern. Our results provide an important benchmark for assessing the impact of COVID-19 on food insecurity.


Intestinal microbiota and probiotics in celiac disease.
Celiac disease (CD) is a common chronic autoimmune enteropathy caused by gluten intake. To date, the only therapy for CD is the complete exclusion of dietary sources of grains and any food containing gluten. It has been hypothesized that the intestinal microbiota is somehow involved in CD. For this reason, probiotics are appearing as an interesting adjuvant in the dietetic management of CD. This review aims to discuss the characteristics of the microbiota in CD subjects and the use of probiotics as a novel therapy for CD. Comparisons between children with CD and controls show that their microbiota profiles differ; the former have fewer lactobacilli and bifidobacteria. Specific probiotics have been found to digest or alter gluten polypeptides. It has also been demonstrated that some bacterial species belonging to the genera Lactobacillus and Bifidobacterium exert protective properties on epithelial cells from damage caused by gliadin.


Inactivating mutations of the histone methyltransferase gene EZH2 in myeloid disorders.
Abnormalities of chromosome 7q are common in myeloid malignancies, but no specific target genes have yet been identified. Here, we describe the finding of homozygous EZH2 mutations in 9 of 12 individuals with 7q acquired uniparental disomy. Screening of a total of 614 individuals with myeloid disorders revealed 49 monoallelic or biallelic EZH2 mutations in 42 individuals; the mutations were found most commonly in those with myelodysplastic/myeloproliferative neoplasms (27 out of 219 individuals, or 12%) and in those with myelofibrosis (4 out of 30 individuals, or 13%). EZH2 encodes the catalytic subunit of the polycomb repressive complex 2 (PRC2), a highly conserved histone H3 lysine 27 (H3K27) methyltransferase that influences stem cell renewal by epigenetic repression of genes involved in cell fate decisions. EZH2 has oncogenic activity, and its overexpression has previously been causally linked to differentiation blocks in epithelial tumors. Notably, the mutations we identified resulted in premature chain termination or direct abrogation of histone methyltransferase activity, suggesting that EZH2 acts as a tumor suppressor for myeloid malignancies.


Diagnostic criteria for multiple sclerosis: 2010 revisions to the McDonald criteria.
New evidence and consensus has led to further revision of the McDonald Criteria for diagnosis of multiple sclerosis. The use of imaging for demonstration of dissemination of central nervous system lesions in space and time has been simplified, and in some circumstances dissemination in space and time can be established by a single scan. These revisions simplify the Criteria, preserve their diagnostic sensitivity and specificity, address their applicability across populations, and may allow earlier diagnosis and more uniform and widespread use.


Age-dependent levels of 5-methyl-, 5-hydroxymethyl-, and 5-formylcytosine in human and mouse brain tissues.
The absolute levels of 5-hydroxymethylcytosine (hmC) and 5-methylcytosine (mC) in human brain tissues at various ages were determined. Additionally, absolute levels of 5-formylcytosine (fC) in adult individuals and cytosine modification levels in sorted neurons were quantified. These data were compared with age-related fC, hmC, and mC levels in mouse brain samples. For hmC, an initial steady increase is observed, which levels off with age to a final steady-state value of 1.2 % in human brain tissue. This level is nearly twice as high as in mouse cerebral cortex. In contrast, fC declines rapidly with age during early developmental stages, thus suggesting that while hmC is a stable epigenetic mark, fC is more likely an intermediate of active DNA demethylation during early brain development. The trends in global cytosine modification dynamics during the lifespan of an organism are conserved between humans and mice and show similar patterns in different organs.


Are Middle-Aged Men with Chronic Kidney Disease at Higher Risk of Having Nocturia than Age-Matched Controls.
We compared the lower urinary tract symptoms between middle-aged men with and without chronic kidney disease (CKD) under 50 years of age. Between October 2010 and July 2013, patients with CKD aged below 50 who received regular follow-ups at the nephrology outpatient clinics were enrolled. We also enrolled men aged below 50 years with estimated glomerular filtration rates (eGFR) higher than 60 mL/min per 1.73 m(2) and without history of kidney diseases from the health examination department as the control group. Clinical parameters and laboratory parameters were collected for analysis and comparison. The lower urinary tract symptoms were evaluated using the International Prostate Symptom Score (IPSS). Metabolic syndromes were defined according to the ATP III guidelines. Overall, 50 men with CKD and 187 age-matched men without CKD were enrolled in the study (age: 44.9 ± 6.9 vs. 43.7.0 ± 4.3, P = 0.11). When compared with the age-matched control group, the occurrences of lower urinary tract symptoms were fewer in CKD patients, though not statistically significant. CKD Patients had significantly higher scores of nocturia (1.3 vs. 0.8, P = 0.02) but had significantly less urinary frequency and relatively fewer urgency score. Nocturia was significantly associated with eGFR. Multivariate analysis showed that lower eGFR and overweight were the only two independent risk factors for nocturia (P < 0.01), but was not the case for metabolic syndromes. Middle-aged men with CKD had significantly more nocturia episodes than the age-matched control group, but had lower urinary frequency scores. Lower eGFR and overweight are independent risk factors for nocturia in middle-aged men.


Traumatic mechanisms and therapeutic results of the perilunate injuries.
In this study, eighteen patients who have had perilunate injury with dislocation or fracture, were evaluated and the patho-mechanics and surgical treatment were studied. According to the Green and O'Brien's classification, type 1 injury occurred in one patient, type 2 in nine, type 4A in five, type 4C in one and type 4D in one. The other one case could not be classified using the Green and O'Brien's criteria. According to the Evans scoring system, good results were achieved in thirteen patients, fair results, in four, poor results in one and very poor results in none. When the patho-mechanics was estimated based on the Mayfield's criteria, seventeen patients were classified as stage 3. However only one case was extremely unusual and should be classified as a subtype of stage 2. The results were good in the majority of patients who had repair of perilunate injury.


Retrofitting BACs with G418 resistance, luciferase, and oriP and EBNA-1 - new vectors for in vitro and in vivo delivery.
Bacterial artificial chromosomes (BACs) have been used extensively for sequencing the human and mouse genomes and are thus readily available for most genes. The large size of BACs means that they can generally carry intact genes with all the long range controlling elements that drive full levels of tissue-specific expression. For gene expression studies and gene therapy applications it is useful to be able to retrofit the BACs with selectable genes such as G418 resistance, reporter genes such as luciferase, and oriP/EBNA-1 from Epstein Barr virus which allows long term episomal maintenance in mammalian cells. We describe a series of retrofitting plasmids and a protocol for in vivo loxP/Cre recombination. The vector pRetroNeo carries a G418 resistance cassette, pRetroNeoLuc carries G418 resistance and a luciferase expression cassette, pRetroNeoLucOE carries G418 resistance, luciferase and an oriP/EBNA-1 cassette and pRetroNeoOE carries G418 resistance and oriP/EBNA-1. These vectors can be efficiently retrofitted onto BACs without rearrangement of the BAC clone. The luciferase cassette is expressed efficiently from the retrofitting plasmids and from retrofitted BACs after transient transfection of B16F10 cells in tissue culture and after electroporation into muscles of BALB/c mice in vivo. We also show that a BAC carrying GFP, oriP and EBNA-1 can be transfected into B16F10 cells with Lipofectamine 2000 and can be rescued intact after 5 weeks. The pRetro vectors allow efficient retrofitting of BACs with G418 resistance, luciferase and/or oriP/EBNA-1 using in vivo expression of Cre. The luciferase reporter gene is expressed after transient transfection of retrofitted BACs into cells in tissue culture and after electroporation into mouse muscle in vivo. OriP/EBNA-1 allows stable maintenance of a 150-kb BAC without rearrangement for at least 5 weeks.


The effects of vibration loading on adipose stem cell number, viability and differentiation towards bone-forming cells.
Mechanical stimulation is an essential factor affecting the metabolism of bone cells and their precursors. We hypothesized that vibration loading would stimulate differentiation of human adipose stem cells (hASCs) towards bone-forming cells and simultaneously inhibit differentiation towards fat tissue. We developed a vibration-loading device that produces 3g peak acceleration at frequencies of 50 and 100 Hz to cells cultured on well plates. hASCs were cultured using either basal medium (BM), osteogenic medium (OM) or adipogenic medium (AM), and subjected to vibration loading for 3 h d(-1) for 1, 7 and 14 day. Osteogenesis, i.e. differentiation of hASCs towards bone-forming cells, was analysed using markers such as alkaline phosphatase (ALP) activity, collagen production and mineralization. Both 50 and 100 Hz vibration frequencies induced significantly increased ALP activity and collagen production of hASCs compared with the static control at 14 day in OM. A similar trend was detected for mineralization, but the increase was not statistically significant. Furthermore, vibration loading inhibited adipocyte differentiation of hASCs. Vibration did not affect cell number or viability. These findings suggest that osteogenic culture conditions amplify the stimulatory effect of vibration loading on differentiation of hASCs towards bone-forming cells.


Synthesis, Structure, and Bonding for Bis(permethylpentalene)diiron.
The synthesis of the first homoleptic double metallocene complex of iron, Fe2Pn*2 (Pn* = permethylpentalene, C8Me6) is described. The structural and electronic properties of Fe2Pn*2 have been characterized by NMR and EPR spectroscopy, single crystal X-ray diffraction, magnetic measurements, cyclic voltammetry, and DFT calculations. Fe2Pn*2 adopts a Ci symmetry in the solid state with a Fe-Fe distance of 2.3175(9) Å, slightly lower than the sum of radii in metallic iron. Magnetic measurements in solution, and of the solid phase between 60 and 300 K, indicate that Fe2Pn*2 is a triplet (S = 1) paramagnet, with effective magnetic moments (μeff) of 3.4 and 3.48 μB, respectively. DFT calculations indicate the origin of this high magnetic moment is likely to be unquenched orbital angular momentum contributions from two SOMOs which have metal d character. Cyclic voltammetry studies demonstrate that Fe2Pn*2 can access four charge states (-1, 0, +1, +2).


Estrogen treatment in multiple sclerosis.
Currently available treatments for multiple sclerosis (MS) reduce inflammatory lesions on MRI and decrease clinical relapses but have limited effects on disability. Novel treatment options that target both the inflammatory as well as the neurodegenerative component of the disease are therefore needed. A growing body of evidence from basic science and clinical studies supports the therapeutic potential of estrogens in MS. Mechanisms of action include both immunomodulatory and directly neuroprotective pathways. A first pilot trial of oral estriol treatment showed encouraging results. There are now several phase II trials underway to further determine the efficacy of estrogen treatment in MS.


Breath-based meditation: A mechanism to restore the physiological and cognitive reserves for optimal human performance.
Stress can be associated with many physiological changes resulting in significant decrements in human performance. Due to growing interests in alternative and complementary medicine by Westerners, many of the traditions and holistic yogic breathing practices today are being utilized as a measure for healthier lifestyles. These state-of-the-art practices can have a significant impact on common mental health conditions such as depression and generalized anxiety disorder. However, the potential of yogic breathing on optimizing human performance and overall well-being is not well known. Breathing techniques such as alternate nostril, Sudarshan Kriya and bhastrika utilizes rhythmic breathing to guide practitioners into a deep meditative state of relaxation and promote self-awareness. Furthermore, yogic breathing is physiologically stimulating and can be described as a natural "technological" solution to optimize human performance which can be categorized into: (1) cognitive function (i.e., mind, vigilance); and (2) physical performance (i.e., cardiorespiratory, metabolism, exercise, whole body). Based on previous studies, we postulate that daily practice of breathing meditation techniques play a significant role in preserving the compensatory mechanisms available to sustain physiological function. This preservation of physiological function may help to offset the time associated with reaching a threshold for clinical expression of chronic state (i.e., hypertension, depression, dementia) or acute state (i.e., massive hemorrhage, panic attic) of medical conditions. However, additional rigorous biomedical research is needed to evaluate the physiological mechanisms of various forms of meditation (i.e., breath-based, mantra, mindfulness) on human performance. These efforts will help to define how compensatory reserve mechanisms of cardiovascular and immune systems are modulated by breath-based meditation. While it has been suggested that breath-based meditation is easier for beginning practitioners when compared to other forms of meditation more research is needed to elucidate these observations. A breath-based meditation sequence such as Sudarshan Kriya has the potential to help develop an individual's self-awareness and support better integration of the brain (i.e., mind) with other organ systems (i.e., body) for enhanced human performance.


Survival in long-term haemodialysis patients: results from the annual survey of the Japanese Society for Dialysis Therapy.
The prognosis for haemodialysis patients is reported to be more favourable in Japan than in Europe or North America. Consequently, evaluation of the death predictors for haemodialysis patients in Japan is of considerable interest outside Japan. The Patient Registration Committee of the Japanese Society for Dialysis Therapy annually surveys the individual patient case mix, laboratory data and important events occurring in the previous years. Thus, using case mix data and laboratory data (including Kt/V and protein catabolic rate) from the 1993 questionnaire survey and the individual patients' life/death statistics from the 1994 questionnaire survey, a logistic regression analysis was conducted on 53867 patients. The analysis indicated that important death risk predictors were: (i) advanced age, (ii) occurrence of diabetes mellitus, (iii) male sex, (iv) Kt/V lower than 1.8, (v) haemodialysis time less than 5 h, (vi) protein catabolic rate less than 0.9 g/kg/day, and (vii) percentage body weight decrease less than 4% and more than 8% during the first haemodialysis session of the week.


Reactive Compression Molding Post-Inverse Vulcanization: A Method to Assemble, Recycle, and Repurpose Sulfur Polymers and Composites.
Inverse vulcanization provides dynamic and responsive materials made from elemental sulfur and unsaturated cross-linkers. These polymers have been used in a variety of applications such as energy storage, infrared optics, repairable materials, environmental remediation, and precision fertilizers. In spite of these advances, there is a need for methods to recycle and reprocess these polymers. In this study, polymers prepared by inverse vulcanization are shown to undergo reactive compression molding. In this process, the reactive interfaces of sulfur polymers are brought into contact by mechanical compression. Upon heating these molds at relatively low temperatures (≈100 °C), chemical bonding occurs at the polymer interfaces by S-S metathesis. This method of processing is distinct from previous studies on inverse vulcanization because the polymers examined in this study do not form a liquid phase when heated. Neither compression nor heating alone was sufficient to mold these polymers into new architectures, so this is a new concept in the manipulation of sulfur polymers. Additionally, high-level ab initio calculations revealed that the weakest S-S bond in organic polysulfides decreases linearly in strength from a sulfur rank of 2 to 4, but then remains constant at about 100 kJ mol-1 for higher sulfur rank. This is critical information in engineering these polymers for S-S metathesis. Guided by this insight, polymer repair, recycling, and repurposing into new composites was demonstrated.


Modulation of respiratory network activity by forelimb and hindlimb locomotor generators.
Early at the onset of exercise, breathing rate accelerates in order to anticipate the increasing metabolic demand resulting from the extra effort produced. Accordingly, the respiratory neural networks are the target of various input signals originating either centrally or peripherally. For example, during locomotion, the activation of muscle sensory afferents is able to entrain and thereby increase the frequency of spontaneous respiratory rhythmogenesis. Moreover, the lumbar spinal networks engaged in generating hindlimb locomotor rhythms are also capable of activating the medullary respiratory generators through an ascending excitatory command. However, in the context of quadrupedal locomotion, the influence of other spinal cord regions, such as cervical and thoracic segments, remains unknown. Using isolated brainstem-spinal cord preparations from neonatal rats and mice, we show that cervicothoracic circuitry may also contribute to locomotion-induced acceleration of respiratory cycle frequency. As previously observed for the hindlimb CPGs, the pharmacological activation of forelimb locomotor networks produces episodes of fictive locomotion that in turn increase the ongoing respiratory rhythm. Thoracic neuronal circuitry may also participate indirectly in this modulation via the activation of both cervical and lumbar CPG neurons. Furthermore, using light stimulation of CHR2-expressing glutamatergic neurons, we found that the modulation of the respiratory rate during locomotion involves lumbar glutamatergic circuitry. Our results demonstrate that during locomotion, the respiratory rhythm-generating networks receive excitatory ascending inputs from the spinal circuits responsible for generating and coordinating fore- and hindlimb movements. This constitutes a distributed central mechanism that contributes to matching breathing rate to the speed of locomotion.


Learning Interpretable Rules for Scalable Data Representation and Classification.
Rule-based models, e.g., decision trees, are widely used in scenarios demanding high model interpretability for their transparent inner structures and good model expressivity. However, rule-based models are hard to optimize, especially on large data sets, due to their discrete parameters and structures. Ensemble methods and fuzzy/soft rules are commonly used to improve performance, but they sacrifice the model interpretability. To obtain both good scalability and interpretability, we propose a new classifier, named Rule-based Representation Learner (RRL), that automatically learns interpretable non-fuzzy rules for data representation and classification. To train the non-differentiable RRL effectively, we project it to a continuous space and propose a novel training method, called Gradient Grafting, that can directly optimize the discrete model using gradient descent. A novel design of logical activation functions is also devised to increase the scalability of RRL and enable it to discretize the continuous features end-to-end. Exhaustive experiments on ten small and four large data sets show that RRL outperforms the competitive interpretable approaches and can be easily adjusted to obtain a trade-off between classification accuracy and model complexity for different scenarios.


Deconstructing Olfactory Stem Cell Trajectories at Single-Cell Resolution.
A detailed understanding of the paths that stem cells traverse to generate mature progeny is vital for elucidating the mechanisms governing cell fate decisions and tissue homeostasis. Adult stem cells maintain and regenerate multiple mature cell lineages in the olfactory epithelium. Here we integrate single-cell RNA sequencing and robust statistical analyses with in vivo lineage tracing to define a detailed map of the postnatal olfactory epithelium, revealing cell fate potentials and branchpoints in olfactory stem cell lineage trajectories. Olfactory stem cells produce support cells via direct fate conversion in the absence of cell division, and their multipotency at the population level reflects collective unipotent cell fate decisions by single stem cells. We further demonstrate that Wnt signaling regulates stem cell fate by promoting neuronal fate choices. This integrated approach reveals the mechanisms guiding olfactory lineage trajectories and provides a model for deconstructing similar hierarchies in other stem cell niches.


Dynamics of dissolution and diffusion-controlled drug release systems.
Analytical expressions were derived to explain the influence of the dissolution/diffusion number (Di) on the time constant and steady-state flux when dispersed drugs are released from a finite matrix. A key novelty of this work is the introduction of a single time-constant that combined the analysis of both dissolution- and diffusion-based systems. Focus is placed on systems with a constant dissolution rate and diffusion coefficient. Solutions, based on the residue theorem, were in agreement with published results describing the transport of estradiol in a polymeric matrix. The experimental cumulative amount of drug released was 0.1 mg/cm² in 100 hours compared to 0.084 mg/cm² predicted by the theoretical model. The process time constant, estimated from the first eigenvalue (t0) and a more accurate statistical approach (t(eff)), showed a consistent decrease with increasing Di values. For a dissolution/diffusion number of 0.21, t0 and t(eff) were estimated at 58.44 and 73.02 hrs, respectively. With the presence of a skin layer, t(eff) increased to 575.7 hrs. These results can be used to assess the relative impact of dissolution and diffusion on the time it takes drugs to attain a therapeutic level in the bloodstream.


Coordinated hydrolysis explains the mechanical behavior of kinesin.
The two-headed motor protein kinesin hydrolyzes nucleotide to move unidirectionally along its microtubule track at speeds up to 1000 nm/s (Saxton et al., 1988) and develops forces in excess of 5 pN (Hunt et al., 1994; Svoboda et al., 1994a). Individual kinesin molecules have been studied recently in vitro, and their behavior has been characterized in terms of force-velocity curves and variance measurements (Svoboda and Block, 1994a; Svoboda et al., 1994b). We present a model for force generation in kinesin in which the ATP hydrolysis reactions are coordinated with the relative positions of the two heads. The model explains the experimental data and permits us to study the relative roles of Brownian motion and elastic deformation in the motor mechanism of kinesin.


Vertically aligned laser sliced MWCNTs.
Applications of multi-walled carbon nanotubes (MWCNTs) benefit from the availability of specific lengths of the material while keeping the outer walls pristine, for example, for applications requiring vertically aligned tubes. To this end, a simple and effective continuous flow 'top down' process to control the length of sliced MWCNTs has been developed using a vortex fluidic device (VFD) coupled with a 1064 nm pulse laser, with the process in the absence of chemicals and any auxiliary substances. Three different length distributions of the sliced MWCNTs, centered at 75 ± 2.1 nm, 300 ± 1.8 nm and 550 ± 1.4 nm, have been generated with the length depending on the VFD operating parameters and laser energy, with the processing resulting in a decrease in side wall defects of the material. We also show the ability to vertically self assemble short MWCNTs on a silicon substrate with control of the surface density coverage using a simple dipping and rinsing method.


Differences in DNA damage produced by incorporation of 5-aza-2'-deoxycytidine or 5,6-dihydro-5-azacytidine into DNA of mammalian cells.
The effects of 5-aza-2'-deoxycytidine (aza-dCyd) and 5,6-dihydro-5-azacytidine (H2-aza-Cyd) on the integrity of DNA from several mammalian cell lines were compared using the alkaline elution technique. While both compounds have been shown to inhibit DNA methylation, a direct comparison of their effects on DNA structure has not previously been reported. Exposure of L1210 cells to H2-aza-Cyd (1-100 micrograms/ml) and simultaneous labeling with [14C]thymidine for 24 h resulted in the production of single-strand breaks in DNA, which were significantly repaired when cells were incubated in drug-free medium for an additional 24 h. This differed from our previous findings for aza-dCyd, confirmed here in parallel experiments, which showed that this compound produces alkali-labile lesions that persist for 48 h. The DNA effects of both drugs were significantly reduced when cells were prelabeled with [14C]thymidine, indicating that production of DNA lesions requires incorporation of the anomalous base. Studies utilizing pulse-labeled DNA indicated that aza-dCyd has little effect on the rate of DNA elongation, whereas H2-aza-Cyd produced a complete inhibition for at least 6 h after drug removal. The contrasting pattern of DNA damage induced by these compounds in L1210 was also observed in two human lymphoblastoid cells lines, one of which was derived from a patient with xeroderma pigmentosum. We had previously concluded that alkali-labile sites in DNA from aza-dCyd-treated cells probably arise due to the chemical instability of aza-dCyd. In contrast, incorporated H2-aza-Cyd is chemically stable. The single-strand breaks produced in H2-aza-Cyd treated cells were not of the alkali-labile type, and may represent an accumulation of DNA replication fragments and/or intermediates in an excision repair process. Thus, the DNA lesions produced by the two drugs have markedly different characteristics, and H2-aza-Cyd should not be considered to be merely a stable pharmacological congener of aza-dCyd.


Relaxation to authentic nitric oxide and SIN-1 in rat isolated mesenteric arteries: variable role for smooth muscle hyperpolarization.
Authentic nitric oxide (NO; 0.1 - 10 micromoles) caused transient, dose-dependent relaxation of phenylephrine-induced tone without changing membrane potential in mesenteric arteries. Larger doses, above 10 micromoles, did not evoke more relaxation (maximal relaxation to 150 micromoles NO in denuded arteries, 69+/-7%, n=8) but stimulated muscle hyperpolarization (maximum 19+/-3 mV, n=5). The soluble guanylyl cyclase inhibitor, 1H-[1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one (ODQ; 10 microM), abolished relaxation to low doses of NO (n=4), but did not modify hyperpolarization with higher doses of NO (n=4). The potassium channel blocker charybdotoxin (ChTX; 50 nM) abolished hyperpolarization to high doses of NO and significantly reduced the maximal relaxation (to 43+/-6%, n=4; P<0.01). ODQ and ChTX together abolished tension and membrane potential change to all doses of NO (n=4). All relaxations to 3-morpholino-sydnonimine (SIN-1; 0.01 - 10 microM) were associated with hyperpolarization. When the endothelium was intact, ChTX inhibited hyperpolarization and relaxation to SIN-1 (n=5), while iberiotoxin (IbTX; 50 nM) or 4-aminopyridine (4-AP; 500 microM) reduced relaxation by 40% and 20%, respectively and by 80% in combination (n=6 in each case). In denuded arteries, relaxation to SIN-1 was unaffected by either ChTX or ODQ alone, but abolished by the inhibitors together (n=6). Alone, 4-AP did not alter relaxation, but in the presence of ODQ it reduced the maximal response by around 45% (n=6; P<0.01). 4-AP, ODQ and IbTX together inhibited relaxation to SIN-1 by 75% (n=6; P<0.01). Therefore, cyclic guanosine 3',5'-monophosphate (cyclic GMP)-independent smooth muscle hyperpolarization, possibly involving direct activation of calcium-activated and voltage-sensitive potassium channels, contributes to relaxation evoked by authentic NO and SIN-1. However, the importance of each pathway depends on the source of NO and with SIN-1 the relative contribution from each pathway is modified by the endothelium.


PEG branched polymer for functionalization of nanomaterials with ultralong blood circulation.
Nanomaterials have been actively pursued for biological and medical applications in recent years. Here, we report the synthesis of several new poly(ethylene glycol) grafted branched polymers for functionalization of various nanomaterials including carbon nanotubes, gold nanoparticles (NPs), and gold nanorods (NRs), affording high aqueous solubility and stability for these materials. We synthesize different surfactant polymers based upon poly(gamma-glutamic acid) (gammaPGA) and poly(maleic anhydride-alt-1-octadecene) (PMHC18). We use the abundant free carboxylic acid groups of gammaPGA for attaching lipophilic species such as pyrene or phospholipid, which bind to nanomaterials via robust physisorption. Additionally, the remaining carboxylic acids on gammaPGA or the amine-reactive anhydrides of PMHC18 are then PEGylated, providing extended hydrophilic groups, affording polymeric amphiphiles. We show that single-walled carbon nanotubes (SWNTs), Au NPs, and NRs functionalized by the polymers exhibit high stability in aqueous solutions at different pH values, at elevated temperatures, and in serum. Moreover, the polymer-coated SWNTs exhibit remarkably long blood circulation (t(1/2) = 22.1 h) upon intravenous injection into mice, far exceeding the previous record of 5.4 h. The ultralong blood circulation time suggests greatly delayed clearance of nanomaterials by the reticuloendothelial system (RES) of mice, a highly desired property for in vivo applications of nanomaterials, including imaging and drug delivery.


Terrorism in South Korea.
South Korea has experienced > 30 suspected terrorism-related events since 1958, including attacks against South Korean citizens in foreign countries. The most common types of terrorism used have included bombings, shootings, hijackings, and kidnappings. Prior to 1990, North Korea was responsible for almost all terrorism-related events inside of South Korea, including multiple assassination attempts on its presidents, regular kidnappings of South Korean fisherman, and several high-profile bombings. Since 1990, most of the terrorist attacks against South Korean citizens have occurred abroad and have been related to the emerging worldwide pattern of terrorism by international terrorist organizations or deranged individuals. The 1988 Seoul Olympic Games provided a major stimulus for South Korea to develop a national emergency response system for terrorism-related events based on the participation of multiple ministries. The 11 September 2001 World Trade Center and Pentagon attacks and the 2001 United States of America (US) anthrax letter attacks prompted South Korea to organize a new national system of emergency response for terrorism-related events. The system is based on five divisions for the response to specific types of terrorist events, involving conventional terrorism, bioterrorism, chemical terrorism, radiological terrorism, and cyber-terrorism. No terrorism-related events occurred during the 2002 World Cup and Asian Games held in South Korea. The emergency management of terrorism-related events in South Korea is adapting to the changing risk of terrorism in the new century.


Low-Cost Particulate Matter Sensors for Monitoring Residential Wood Burning.
Conventional monitoring systems for air quality, such as reference stations, provide reliable pollution data in urban settings but only at relatively low spatial density. This study explores the potential of low-cost sensor systems (LCSs) deployed at homes of residents to enhance the monitoring of urban air pollution caused by residential wood burning. We established a network of 28 Airly (Airly-GSM-1, SP. Z o.o., Poland) LCSs in Kristiansand, Norway, over two winters (2021-2022). To assess performance, a gravimetric Kleinfiltergerät measured the fine particle mass concentration (PM2.5) in the garden of one participant's house for 4 weeks. Results showed a sensor-to-reference correlation equal to 0.86 for raw PM2.5 measurements at daily resolution (bias/RMSE: 9.45/11.65 μg m-3). High-resolution air quality maps at a 100 m resolution were produced by combining the output of an air quality model (uEMEP) using data assimilation techniques with the network data that were corrected and calibrated by using a proposed five-step network data processing scheme. Leave-one-out cross-validation demonstrated that data assimilation reduced the model's RMSE, MAE, and bias by 44-56, 38-48, and 41-52%, respectively.


Silencing of carbonic anhydrase in an Anopheles gambiae larval cell line, Ag55.
RNAi has been used extensively to down-regulate proteins in adult mosquitoes; however, it is not well adapted for use in larvae. Larval mosquitoes can generate a pH as high as 10.5 in the anterior region of their midgut. The mechanisms responsible for the generation and maintenance of this pH are not entirely understood, but members of the carbonic anhydrase (CA) family of enzymes have been implicated. Here we use an An. gambiae larval cell line, Ag55 cells, to demonstrate that application of full-length double-stranded RNA specific to one CA, AgCA9, is sufficient to silence AgCA9 mRNA and down-regulate the corresponding protein. This is a first step towards determining the role(s) of these enzymes in pH regulation.


Outcome Expectations and Osteoarthritis: Association of Perceived Benefits of Exercise With Self-Efficacy and Depression.
Outcome expectancy is recognized as a determinant of exercise engagement and adherence. However, little is known about which factors influence outcome expectations for exercise among people with knee osteoarthritis (OA). This is the first study to examine the association of outcome expectations for exercise with demographic, physical, and psychosocial outcomes in individuals with knee OA. We performed a cross-sectional analysis of the baseline data from a randomized trial of tai chi versus physical therapy in participants with symptomatic knee OA. Knee pain was evaluated using the Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC). Outcome expectations for exercise, self-efficacy, depression, anxiety, stress, and social support were measured using standard instruments. Logistic regression models were utilized to determine associations with outcome expectations. There were 262 participants, with a mean age of 59.8 years and a mean body mass index of 32.1 kg/m2 ; 69.1% of the participants were female, 51.5% were white, the mean disease duration was 8.6 years, and the mean WOMAC knee pain and function scores were 260.8 and 906.8, respectively. Higher outcome expectations for exercise were associated with greater self-efficacy (odds ratio [OR] 1.25 [95% confidence interval (95% CI) 1.11-1.41]; P = 0.0004), as well as with fewer depressive symptoms (OR 0.84 for each 5-point increase [95% CI 0.73-0.97]; P = 0.01). Outcome expectancy was not significantly associated with sex, race, education, pain, function, radiographic severity, social support, anxiety, or stress. Our results suggest significant associations between outcome expectations for exercise and self-efficacy and depression. Future studies should examine how these relationships longitudinally affect long-term clinical outcomes of exercise-based treatment for knee OA.


Chimpanzees flexibly update working memory contents and show susceptibility to distraction in the self-ordered search task.
Working memory (WM) is a core executive function that allows individuals to hold, process and manipulate information. WM capacity has been repeatedly nominated as a key factor in human cognitive evolution; nevertheless, little is known about the WM abilities of our closest primate relatives. In this study, we examined signatures of WM ability in chimpanzees (Pan troglodytes). Standard WM tasks for humans (Homo sapiens) often require participants to continuously update their WM. In Experiment 1, we implemented this updating requirement in a foraging situation: zoo-housed chimpanzees (n = 13) searched for food in an array of containers. To avoid redundant searches, they needed to continuously update which containers they had already visited (similar to WM paradigms for human children) with 15 s retention intervals in between each choice. We examined chimpanzees' WM capacity and to what extent they used spatial cues and object features to memorize their previous choices. In Experiment 2, we investigated how susceptible their WM was to attentional interference, an important signature, setting WM in humans apart from long-term memory. We found large individual differences with some individuals remembering at least their last four choices. Chimpanzees used a combination of spatial cues and object features to remember which boxes they had chosen already. Moreover, their performance decreased specifically when competing memory information was introduced. Finally, we found that individual differences in task performance were highly reliable over time. Together, these findings show remarkable similarities between human and chimpanzee WM abilities despite evolutionary and life-history differences.


[Acupuncture for premature ovarian failure: a prospective cohort study].
To explore the clinical efficacy and action mechanism of acupuncture for premature ovarian failure (POF). According to prospective cohort study design, 30 cases were included. Based on theory of regulating Chong and Conception Vessels as well as soothing the liver and calming the nerves, acupuncture at Guanyuan (CV 4), Dahe (KI 12), Taixi (KI 3), Taichong (LR 3), Baihui (GV 20) was applied, three times per week and 3 months were considered as one session. Totally two sessions were performed. The menstruation condition, estradiol (E2), follicle-stimulating hormone (FSH) and scores of the clinical perimenopausal symptoms were taken as treatment outcomes. The total effective rate was 86.7% (26/30) and the cured rate was 16.7% (5/30). The recovery rate of menstruation was 16.7% (5/30) and the regain rate of menstruation was 56.7% (17/30). After the treatment, the level of E2 was significantly increased from (45.41 +/- 18. 09) pmol/L to (59.07 +/- 24.21) pmol/L (P < 0.05), and the score of perimenopausal symptoms was obviously reduced from 14.28 +/- 8. 30 to 4.04 +/- 3.28 (P < 0.01). However, no statistical differences could be seen in FSH before and after treatment (P > 0.05). Acupuncture has certain effect on improving menstruation and perimenopausal symptoms in POF patients, which is likely to he related with increasing the level of E2.


MicroRNAs contribute to compensatory β cell expansion during pregnancy and obesity.
Pregnancy and obesity are frequently associated with diminished insulin sensitivity, which is normally compensated for by an expansion of the functional β cell mass that prevents chronic hyperglycemia and development of diabetes mellitus. The molecular basis underlying compensatory β cell mass expansion is largely unknown. We found in rodents that β cell mass expansion during pregnancy and obesity is associated with changes in the expression of several islet microRNAs, including miR-338-3p. In isolated pancreatic islets, we recapitulated the decreased miR-338-3p level observed in gestation and obesity by activating the G protein-coupled estrogen receptor GPR30 and the glucagon-like peptide 1 (GLP1) receptor. Blockade of miR-338-3p in β cells using specific anti-miR molecules mimicked gene expression changes occurring during β cell mass expansion and resulted in increased proliferation and improved survival both in vitro and in vivo. These findings point to a major role for miR-338-3p in compensatory β cell mass expansion occurring under different insulin resistance states.


Living systematic reviews: 2. Combining human and machine effort.
New approaches to evidence synthesis, which use human effort and machine automation in mutually reinforcing ways, can enhance the feasibility and sustainability of living systematic reviews. Human effort is a scarce and valuable resource, required when automation is impossible or undesirable, and includes contributions from online communities ("crowds") as well as more conventional contributions from review authors and information specialists. Automation can assist with some systematic review tasks, including searching, eligibility assessment, identification and retrieval of full-text reports, extraction of data, and risk of bias assessment. Workflows can be developed in which human effort and machine automation can each enable the other to operate in more effective and efficient ways, offering substantial enhancement to the productivity of systematic reviews. This paper describes and discusses the potential-and limitations-of new ways of undertaking specific tasks in living systematic reviews, identifying areas where these human/machine "technologies" are already in use, and where further research and development is needed. While the context is living systematic reviews, many of these enabling technologies apply equally to standard approaches to systematic reviewing.


Increasing Hepatitis C treatment uptake among HIV-infected patients using an HIV primary care model.
Access to Hepatitis C (HCV) care is low among HIV-infected individuals, highlighting the need for new models to deliver care for this population. Retrospective cohort analysis that compared the number of HIV patients who initiated HCV therapy: hepatology (2005-2008) vs. HIV primary care model (2008-2011). Logistic-regression modeling was used to ascertain factors associated with HCV therapy initiation and achievement of sustained viral response (SVR). Of 196 and 163 patients that were enrolled in the HIV primary care and hepatology models, 48 and 26 were treated for HCV, respectively (p = 0.043). The HIV/HCV-patient referral rate did not differ during the two study periods (0.10 vs. 0.12/patient-yr, p = 0.18). In unadjusted analysis, predictors (p < 0.05) of HCV treatment initiation included referral to the HIV primary care model (OR: 1.7), a CD4+ count ≥400/mm3 (OR: 1.8) and alanine aminotranferase level ≥63U/L (OR: 1.9). Prior psychiatric medication use correlated negatively with HCV treatment initiation (OR: 0.6, p = 0.045). In adjusted analysis the strongest predictor of HCV treatment initiation was CD4+ count (≥400/mm3, OR: 2.1, p = 0.01). There was no significant difference in either clinic model (primary care vs. hepatology) in the rates of treatment discontinuation due to adverse events (29% vs. 16%), loss to follow-up (8 vs. 8%), or HCV SVR (44 vs. 35%). Using a HIV primary care model increased the number of HIV patients who initiate HCV therapy with comparable outcomes to a hepatology model.


Workplace accommodations for persons with physical disabilities: evidence synthesis of the peer-reviewed literature.
To identify and synthesize research evidence on workplace accommodations used by employers to recruit, hire, retain, and promote persons with physical disabilities. A structured search of six electronic journal databases was undertaken to identify peer-reviewed literature on the topic published from January 1990 to March 2016. Articles describing or evaluating workplace disability accommodation policies and practices were given a full-text review. Topic experts were contacted to identify additional studies. Details on specific accommodations described in 117 articles were synthesized and organized into three groups comprised of a total of 12 categories. The majority of studies did not rigorously evaluate effectiveness or cost-effectiveness of the accommodations under study. This evidence synthesis provides an overview of the peer-reviewed literature of value to occupational rehabilitation professionals and employers seeking guidance on workplace accommodation policies and practices for persons with physical disabilities. A wide range of accommodation options is available for addressing physical, social, and attitudinal barriers to successful employment. Besides physical/technological modifications, accommodations to enhance workplace flexibility and worker autonomy and strategies to promote workplace inclusion and integration are important. More comprehensive reporting and evaluations of the effectiveness of accommodations in research literature are needed to develop best practices for accommodating persons with disabilities. Implications for rehabilitation There is a substantial peer-reviewed literature that provides insights into the barriers for persons with physical disabilities and the workplace accommodation practices to address them, though rigorous evaluations of effectiveness and cost-effectiveness are uncommon. Attitudinal and social barriers stemming from stereotypes, ignorance and lack of knowledge are as important as physical barriers to employment for persons with physical disabilities. In addition to physical/technological modifications, accommodations to enhance workplace flexibility and autonomy of a worker, as well as strategies to promote workplace inclusion and integration may facilitate successful employment of persons with physical disabilities.


Short-forms of the UK WAIS-R: regression equations and their predictive validity in a general population sample.
A sample of 200 healthy subjects, representative of the adult UK population in terms of age, sex and social class distribution, were administered a full-length WAIS-R (UK). Regression equations were built to predict full-length IQ from a series of short-forms. The short-forms ranged from a two-subtest version proposed by Silverstein (1982) to a seven-subtest version proposed by Warrington, James & Maciejewski (1986). Regression equations, their standard errors of estimate and confidence intervals are presented as well as IQ conversion tables. The short-forms are evaluated in terms of their validity in predicting full-length IQ and in terms of their clinical utility. The advantages of regression-based estimates of full-length IQ over those derived from conventional prorating are discussed.


Normal peroxisomal function and absent skeletal manifestations in Conradi-Hünermann syndrome.
We describe a child with the classic cutaneous and ocular manifestations of Conradi-Hünermann syndrome in whom repeated roentgenographic studies during the first 2 years of life revealed no evidence of epiphyseal stippling. The findings in this case and others from the literature suggest that skeletal changes may be absent or show limited expression in patients with this condition and that chondrodysplasia punctata should not be considered an invariable feature of Conradi-Hünermann syndrome. Studies of peroxisomal function in our patient failed to confirm two previous reports of a significant reduction in activity of the peroxisomal enzyme dihydroxyacetone phosphate acyltransferase in this disorder.


Multiphasic acute disseminated encephalomyelitis and differential with early onset multiple sclerosis.
Multiple sclerosis is considered the most frequent demyelinating disorder of the Central Nervous System (CNS) among young adults, yet is very rare before 10 years old. Acute disseminated encephalomyelitis is a monophasic, polysymptomatic disorder that involves the CNS white matter with demyelinating lesions, which usually occurs after systemic viral infections. These two demyelinating diseases can present initially as an acute focal neurological syndrome and they can be difficult to distinguish. We describe a case of a nine-year-old girl that presented initially with dysphonia, gait ataxia, eyelid myokymia and brainstem disturbances. This was her second episode; the first episode was at the age of four years old. She recovered without neurological sequelae. The brain magnetic resonance imaging (MRI) demonstrated multiple demyelinating lesions in the white matter, cortical regions of the frontal lobe, periventricular distribution, internal capsule, corpus callosum and cerebellum. The purpose of the presentation of this case was to highlight the similarities between these two entities, since the clinical picture and neuroimaging are difficult to distinguish, mainly in relation to the first episode.


Structural disorder in the complex of human pregnane X receptor and the macrolide antibiotic rifampicin.
The human nuclear xenobiotic receptor, pregnane X receptor (PXR), detects a variety of structurally distinct endogenous and xenobiotic compounds and controls expression of genes central to drug and cholesterol metabolism. The macrolide antibiotic rifampicin, a front-line treatment for tuberculosis, is an established PXR agonist and, at 823 Da, is one of the largest known ligands for the receptor. We present the 2.8 A crystal structure of the ligand-binding domain of human PXR in complex with rifampicin. We also use structural and mutagenesis data to examine the origins of the directed promiscuity exhibited by the PXRs across species. Three structurally flexible loops adjacent to the ligand-binding pocket of PXR are disordered in this crystal structure, including the 200-210 region that is part of a sequence insert novel to the promiscuous PXRs relative to other members of the nuclear receptor superfamily. The 4-methyl-1-piperazinyl ring of rifampicin, which would lie adjacent to the disordered protein regions, is also disordered and not observed in the structure. Taken together, our results indicate that one wall of the PXR ligand-binding cavity can remain flexible even when the receptor is in complex with an activating ligand. These observations highlight the key role that structural flexibility plays in PXR's promiscuous response to xenobiotics.


Stepwise dissolution and composition determination of samples of multiple crystals using a dissolution medium containing aqueous alcohol and fluorocarbon phases.
Crystallisations are widely used in pharmaceutical and fine chemical manufacturing to control impurity levels, however crystallisations do not always reduce impurities to acceptable levels. Information on the location and distribution of impurities in crystallised materials would be helpful in such cases. A two phase dissolution medium featuring a fluorocarbon non-solvent vehicle and a aqueous ethanol solvent phase has been used to determine the composition of multi-particle crystalline samples through a partial dissolution approach combined with particle sizing and HPLC analysis. 4-Chloro-2-nitroacetanilide (1) was chosen as the host compound for this study, with 4-methyl-2-nitroacetanilide (2) and 4-tert-butyl-2-nitroacetanilide (3) chosen as the guest impurities that were added to supersaturated toluene solutions of 1 at levels up to 5 mol%. The crystals that formed were subjected to a series of partial dissolution steps carried out using the biphasic dissolution medium composed of a 50% aqueous ethanol solvent phase and a perfluorohexane continuous phase. To inhibit particle agglomeration, the mixture also contained 13,13,14,14,15,15,16,16,17,17,18,18-dodecafluoro-2,5,8,11-tetraoxaoctadecane (4) as a non-ionic surfactant. The partial dissolution steps showed a relatively even dissolution with each sequential step as determined from particle sizing. Analysis of the solutions by HPLC from each partial dissolution step allowed the level of impurity to be determined, and when combined with the particle sizing data this allowed an impurity distribution to be generated. Impurity 2 was found to be relatively evenly distributed while impurity 3 was localised on or near the surfaces of crystals.


Complex transcriptional control of the AZFa gene DDX3Y in human testis.
The human DEAD-box Y (DBY) RNA helicase (aka DDX3Y) gene is thought to be the major azoospermia factor a (AZFa) gene in proximal Yq11. Men with its deletion display no somatic pathologies, but suffer from complete absence of germ cells. Accordingly, DDX3Y protein is expressed only in the germline in spermatogonia, although the transcripts were found in many tissues. Here, we show the complex transcriptional control of a testis-specific DDX3Y transcript class with initiation at different sites upstream of the gene's open reading frame (5'Untranslated Region; UTR) and with polyadenylation in their proximal 3'UTR. The most distal transcriptional start site (TSS; ∼1 kb upstream) was mapped in MSY2, a Y-specific minisatellite. As this testis-specific 5'UTR was subsequently processed by three alternative splicing events, it has been tentatively designated 'exon-T'(estis). The MSY2 sequence unit was also found upstream of the mouse Ddx3y gene. However, only after its tandem amplification on the Y chromosome of Platyrrhini (new world monkeys) and Catarrhini (old world monkeys) did MSY2 become part of a novel distal promoter for DDX3Y expression in testis tissue and provides a second transcriptional start site (T-TSS-II) in Catarrhini. We therefore suggest that the development of a novel distal DDX3Y promoter in primates, which is activated only in testis tissue, is probably part of the gene's germline translation control.


Genome-wide transcription start site mapping of Bradyrhizobium japonicum grown free-living or in symbiosis - a rich resource to identify new transcripts, proteins and to study gene regulation.
Differential RNA-sequencing (dRNA-seq) is indispensable for determination of primary transcriptomes. However, using dRNA-seq data to map transcriptional start sites (TSSs) and promoters genome-wide is a bioinformatics challenge. We performed dRNA-seq of Bradyrhizobium japonicum USDA 110, the nitrogen-fixing symbiont of soybean, and developed algorithms to map TSSs and promoters. A specialized machine learning procedure for TSS recognition allowed us to map 15,923 TSSs: 14,360 in free-living bacteria, 4329 in symbiosis with soybean and 2766 in both conditions. Further, we provide proteomic evidence for 4090 proteins, among them 107 proteins corresponding to new genes and 178 proteins with N-termini different from the existing annotation (72 and 109 of them with TSS support, respectively). Guided by proteomics evidence, previously identified TSSs and TSSs experimentally validated here, we assign a score threshold to flag 14 % of the mapped TSSs as a class of lower confidence. However, this class of lower confidence contains valid TSSs of low-abundant transcripts. Moreover, we developed a de novo algorithm to identify promoter motifs upstream of mapped TSSs, which is publicly available, and found motifs mainly used in symbiosis (similar to RpoN-dependent promoters) or under both conditions (similar to RpoD-dependent promoters). Mapped TSSs and putative promoters, proteomic evidence and updated gene annotation were combined into an annotation file. The genome-wide TSS and promoter maps along with the extended genome annotation of B. japonicum represent a valuable resource for future systems biology studies and for detailed analyses of individual non-coding transcripts and ORFs. Our data will also provide new insights into bacterial gene regulation during the agriculturally important symbiosis between rhizobia and legumes.


Resuscitation with lactated ringer's does not increase inflammatory response in a Swine model of uncontrolled hemorrhagic shock.
Lactated Ringer's (LR) and normal saline (NS) are widely and interchangeably used for resuscitation of trauma victims. Studies show LR to be superior to NS in the physiologic response to resuscitation. Recent in vitro studies demonstrate equivalent effects of LR and NS on leukocytes. We aimed to determine whether LR resuscitation would produce an equivalent inflammatory response compared with normal saline (NS) resuscitation in a clinically relevant swine model of uncontrolled hemorrhagic shock. Thirty-two swine were randomized. Control animals (n = 6) were sacrificed following induction of anesthesia for baseline data. Sham animals (n = 6) underwent laparotomy and 2 h of anesthesia. Uncontrolled hemorrhagic shock animals (n = 10/group) underwent laparotomy, grade V liver injury, and blinded resuscitation with LR or NS to maintain baseline blood pressure for 1.5 h before sacrifice. Lung was harvested, and tissue mRNA levels of interleukin-6 (IL-6), granulocyte colony-stimulating factor (G-CSF), and tumor necrosis factor-alpha (TNF-alpha) were determined using quantitative reverse transcriptase polymerase chain reaction (Q-RT-PCR). Sections of lung were processed and examined for neutrophils sequestered within the alveolar walls. Cytokine analysis showed no difference in IL-6 gene transcription in any group (P = 0.99). Resuscitated swine had elevated G-CSF and TNF-alpha gene transcription, but LR and NS groups were not different from each other (P= 0.96 and 0.10, respectively). Both resuscitation groups had significantly more alveolar neutrophils present than controls (P < 0.01) and shams (P < 0.05) but were not different from one another (P= 0.83). LR and NS resuscitation have equivalent effects on indices of inflammation in the lungs in our model of uncontrolled hemorrhagic shock.


Human constant regions influence the antibody binding characteristics of mouse-human chimeric IgG subclasses.
Although antibody affinity is primarily determined by immunoglobulin variable region structure human IgG antibodies of the four subclasses specific for the same antigen have been shown to differ in their affinity. To explore the influence of the immunoglobulin constant region on functional antibody affinity, a set of V region identical mouse-human chimeric IgG subclasses specific for TAG72 (tumour-associated glycoprotein) were studied. Biomolecular interaction analysis (BIA) was used to determine the binding kinetics of whole IgG subclasses and F(ab')2 fragments. Despite identical V regions, binding kinetics differed for the four subclasses. The apparent dissociation rate constants of the intact immunoglobulins ranked IgG4 < IgG3 < IgG2 < IgG1. In contrast, analysis of the binding characteriztics of the F(ab')2 fragments derived from IgG1, IgG2 and IgG4 revealed identical binding kinetics. The structure of the constant regions of the humanized IgG subclass antibodies clearly influenced functional antibody affinity, as has been described for the murine IgG subclasses. The exact mechanism for this phenomenon remains obscure but such differences should be taken into account when designing or choosing antibodies for therapeutic use.


The growing burden of tuberculosis: global trends and interactions with the HIV epidemic.
The increasing global burden of tuberculosis (TB) is linked to human immunodeficiency virus (HIV) infection. We reviewed data from notifications of TB cases, cohort treatment outcomes, surveys of Mycobacterium tuberculosis infection, and HIV prevalence in patients with TB and other subgroups. Information was collated from published literature and databases held by the World Health Organization (WHO), the Joint United Nations Programme on HIV/Acquired Immunodeficiency Syndrome (UNAIDS), the US Census Bureau, and the US Centers for Disease Control and Prevention. There were an estimated 8.3 million (5th-95th centiles, 7.3-9.2 million) new TB cases in 2000 (137/100,000 population; range, 121/100,000-151/100,000). Tuberculosis incidence rates were highest in the WHO African Region (290/100,000 per year; range, 265/100,000-331/100,000), as was the annual rate of increase in the number of cases (6%). Nine percent (7%-12%) of all new TB cases in adults (aged 15-49 years) were attributable to HIV infection, but the proportion was much greater in the WHO African Region (31%) and some industrialized countries, notably the United States (26%). There were an estimated 1.8 million (5th-95th centiles, 1.6-2.2 million) deaths from TB, of which 12% (226 000) were attributable to HIV. Tuberculosis was the cause of 11% of all adult AIDS deaths. The prevalence of M tuberculosis-HIV coinfection in adults was 0.36% (11 million people). Coinfection prevalence rates equaled or exceeded 5% in 8 African countries. In South Africa alone there were 2 million coinfected adults. The HIV pandemic presents a massive challenge to global TB control. The prevention of HIV and TB, the extension of WHO DOTS programs, and a focused effort to control HIV-related TB in areas of high HIV prevalence are matters of great urgency.


Innovations in surgery simulation: a review of past, current and future techniques.
As a result of recent work-hours limitations and concerns for patient safety, innovations in extraclinical surgical simulation have become a desired part of residency education. Current simulation models, including cadaveric, animal, bench-top, virtual reality (VR) and robotic simulators are increasingly used in surgical training programs. Advances in telesurgery, three-dimensional (3D) printing, and the incorporation of patient-specific anatomy are paving the way for simulators to become integral components of medical training in the future. Evidence from the literature highlights the benefits of including simulations in surgical training; skills acquired through simulations translate into improvements in operating room performance. Moreover, simulations are rapidly incorporating new medical technologies and offer increasingly high-fidelity recreations of procedures. As a result, both novice and expert surgeons are able to benefit from their use. As dedicated, structured curricula are developed that incorporate simulations into daily resident training, simulated surgeries will strengthen the surgeon's skill set, decrease hospital costs, and improve patient outcomes.


Accumulation of polystyrene microplastics in juvenile Eriocheir sinensis and oxidative stress effects in the liver.
As a widespread and ubiquitous pollutant of marine ecosystems, microplastic has the potential to become an emerging global threat for aquatic organisms. The present study aims to elucidate the effects of microplastics on the growth, accumulation and oxidative stress response in the liver of Eriocheir sinensis. Fluorescent microplastic particles (diameter = 0.5 μm) accumulated in the gill, liver and gut tissues of E. sinensis were investigated when crabs were exposed to a concentration of 40000 μg/L for 7 days. A 21 day toxicity test suggested that the rate of weight gain, specific growth rate, and hepatosomatic index of E. sinensis decreased with increasing microplastic concentration (0 μg/L, 40 μg/L, 400 μg/L, 4000 μg/L and 40000 μg/L). The activities of AChE and GPT in crabs exposed to microplastics were lower than those in control group. GOT activity increased significantly after exposure to a low concentration of microplastics and then decreased continuously with increasing microplastic concentrations. The activities of superoxide dismutase (SOD), aspartate transaminase (GOT), glutathione (GSH), and glutathione peroxidase (GPx) increased in specimens exposed to low concentrations of microplastics (40 and 400 μg/L) compared to the control and decreased in organisms exposed to high concentrations (4000 and 40000 μg/L). In contrast, the activities of acetylcholinesterase, catalase (CAT), and alanine aminotransferase were significantly lower in the organisms exposed to microplastics compared to control animals. Upon exposure to increasing microplastic concentrations, the expression of genes encoding the antioxidants SOD, CAT, GPx and glutathione S-transferase in the liver decreased after first increasing. Exposure to microplastics increased the expression of the gene encoding p38 in the MAPK signaling pathway and significantly decreased the expressions of genes encoding ERK, AKT, and MEK. The results of this study demonstrate that microplastics can accumulate in the tissues of E. sinensis and negatively affect growth. In addition, exposure to microplastics causes damage and induces oxidative stress in the hepatopancreas of E. sinensis. The findings provide basic biological data for environmental and human risk assessments of microplastics of high concern.