Recurrence of autoimmune liver disease and inflammatory bowel disease after pediatric liver transplantation.
Approximately 10% of children with autoimmune hepatitis (AIH) and 30% of those with sclerosing cholangitis (SC) require liver transplantation (LT). LT is indicated in patients who present with fulminant hepatic failure (ie, with encephalopathy) and in those who develop end-stage liver disease despite treatment. After LT, recurrent AIH is reported in approximately 30% of patients and recurrent SC in up to 50%. Diagnosis of recurrence is based on biochemical abnormalities, seropositivity for autoantibodies, interface hepatitis on histology, steroid dependence, and, for SC, presence of cholangiopathy. Recurrence of SC after LT is often associated with poorly controlled inflammatory bowel disease (IBD). Recurrence may even appear years after LT; therefore, steroid-based immunosuppression should be maintained at a higher dose than that used for patients transplanted for nonautoimmune liver diseases. Although the impact of recurrent disease on graft function is controversial, it seems that in pediatric LT recipients recurrence of AIH or SC is associated with compromised graft survival. Exacerbation of preexistent IBD may be observed after LT for SC or AIH, and IBD appears to have a more aggressive course than before LT. In addition, IBD can develop de novo following LT. Liver Transplantation 22 1275-1283 2016 AASLD.


Initial Treatment of Cancer Patients with Fluconazole-Susceptible Dose-Dependent Candida glabrata Fungemia: Better Outcome with an Echinocandin or Polyene Compared to an Azole?
The 28-day crude mortality rate in 68 cancer patients with fluconazole-susceptible dose-dependent Candida glabrata fungemia started on treatment (within 48 h after blood culture collection) with an echinocandin or liposomal amphotericin-B was better (30%) than those treated with azole monotherapy (52%) (P = 0.07). After adjusting for confounders, azole monotherapy also was associated with worse 28-day survival (hazard ratio, 3.8; P = 0.003).


[Evaluation of the fluoride concentration and consumption of mineral water].
Considering that water is an importance source of fluoride intake, and that the consumption of mineral water and prevalence of dental fluorosis have been increasing, the aim of this study was to evaluate the consumption of mineral water and its fluoride concentration. The study was performed in residential districts of the municipality of Bauru, State of São Paulo, by means of stratified sampling via clusters. Each cluster corresponded to one residential block. For randomization purposes, the residential blocks were numbered within the 17 districts established by the city plan. One thousand homes were thus visited. Mineral water samples were collected using previously labeled 50 ml plastic flasks. Fluoride analysis was done using an ion-sensitive electrode (Orion 9609), after buffering using TISAB II. Information on the consumption of mineral water was obtained by means of applying a questionnaire. Around 29.72% of the city's population was consuming mineral water. In the 260 samples analyzed from 29 different brands of water, the fluoride concentration ranged from 0.045 to 1.515 mg/l. For one brand, the label stated that the fluoride concentration was 0.220 mg/l, but analysis revealed a concentration of 1.515 mg/l. Moreover, some brands did not specify the fluoride concentration on the label and, for these, the analysis showed concentrations ranging from 0.049 to 0.924 mg/l. The results demonstrated wide variation in fluoride concentrations and reinforce the importance of the control of such waters by the sanitary surveillance agency.


Towards an efficient compression of 3D coordinates of macromolecular structures.
The size and complexity of 3D macromolecular structures available in the Protein Data Bank is constantly growing. Current tools and file formats have reached limits of scalability. New compression approaches are required to support the visualization of large molecular complexes and enable new and scalable means for data analysis. We evaluated a series of compression techniques for coordinates of 3D macromolecular structures and identified the best performing approaches. By balancing compression efficiency in terms of the decompression speed and compression ratio, and code complexity, our results provide the foundation for a novel standard to represent macromolecular coordinates in a compact and useful file format.


New pathogens and old resistance genes.
In the past fifty years the treatment and ecology of infectious diseases has evolved greatly. The major changes have come as the development of "new" pathogens which are usually "old" pathogens in new guises. The chief environmental factor responsible for this change is the use of antimicrobial agents. It is estimated that more than 10(9) kg of antibiotic have been produced and used since the early 1959's. In spite of this threat, bacterial pathogens have not only survived, but even flourished. It is now too late to reverse this trend, since the treatment of infectious diseases relies heavily on antibiotics and there is a well-established pool of antibiotic resistance genes in nature. Hospital are unique, varied ecological niches for microbial diversity, and we know little about where resistance genes come from or the pathways by which these genes are passed around the bacterial population. Since less than 10% of the bacterial species on earth can be identified, we are ignorant of a large component of the factors responsible for health, disease and antibiotic resistance. What can we look forward to? There has been much talk of reemerging pathogens such as Mycobacterium tuberculosis, Listeria monocytogenes, enterococci and various fungi. These organisms have surfaced because of their ability to survive in hostile environments (antibiotics, antiseptics). We should anticipate the reappearance of other such organisms, and clinicians and the pharmaceutical industry should be prepared for this. Now it is the time to get profit from the lessons of the past, which, together with better knowledge of microbial biochemistry and ecology, must be applied to the problems of infectious diseases.


Pentanuclear dysprosium hydroxy cluster showing single-molecule-magnet behavior.
A pentanuclear dysprosium hydroxy cluster of composition [Dy 5(mu 4-OH)(mu 3-OH) 4(mu-eta (2)-Ph 2acac) 4(eta (2)-Ph 2acac) 6] ( 1; Ph 2acac = dibenzoylmethanide) was prepared starting from [DyCl 3.6H 2O] and dibenzoylmethane. Both static (dc) and dynamic (ac) magnetic properties of 1 have been studied. Below 3 K, the appearance of slow relaxation of the magnetization typical for single-molecule magnets is seen, even if no hysteresis effects on the M vs H data are observed above 1.8 K.


Substance Use among Exclusive Electronic Cigarette Users and Dual Combustible Cigarette Users: Extending Work to Adult Users.
Electronic cigarettes (e-cigarettes) have become increasingly popular in recent years. Existing evidence indicates e-cigarettes used in isolation are less harmful than combustible cigarettes, yet emerging work has demonstrated that adults use e-cigarettes largely in combination with combustible cigarettes (i.e. dual use). Despite this data, little is understood about how exclusive and dual e-cigarette users may differ across behavioral outcomes, such as substance use dependence and behaviors among adults. Thus, the current project examined differences in e-cigarette dependence, problematic alcohol use, cannabis use, and nonmedical prescription opioid use (e.g. methadone and oxycodone) across both exclusive and dual e-cigarette adult users. Participants included 531 (53.6% female; Mage = 35.29 years, SD = 10.44) adults with past-month e-cigarette use. Dual users reported greater e-cigarette dependence, alcohol use, current cannabis use, and endorsement of nonprescription opioid use. Conclusions/Importance: Importantly, this work suggests that adults who use both combustible and e-cigarettes may constitute a group more vulnerable to experiencing problematic substance use and more severe e-cigarette dependence than exclusive e-cigarette users.


A unique case of bilateral hydronephrosis caused by a fecalith.
This is a unique case that signifies the importance to look beyond the genitourinary system for causes of hydronephrosis. In addition, we outline the manner in which a fecalith should be addressed.


Iron and Parkinson's disease: chelators to the rescue?
The essential metal iron has long been implicated in the neuronal damage associated with Parkinson's disease. Recent findings show that iron chelation may prevent the reductions in dopamine and motor disturbances associated with this disease, and suggest the need to examine the role of dietary iron and the use of metal chelators in neurodegenerative disorders.


High process yield rates of thermoplastic nanofluidic devices using a hybrid thermal assembly technique.
Over the past decade, thermoplastics have been used as alternative substrates to glass and Si for microfluidic devices because of the diverse and robust fabrication protocols available for thermoplastics that can generate high production rates of the desired structures at low cost and with high replication fidelity, the extensive array of physiochemical properties they possess, and the simple surface activation strategies that can be employed to tune their surface chemistry appropriate for the intended application. While the advantages of polymer microfluidics are currently being realized, the evolution of thermoplastic-based nanofluidic devices is fraught with challenges. One challenge is assembly of the device, which consists of sealing a cover plate to the patterned fluidic substrate. Typically, channel collapse or substrate dissolution occurs during assembly making the device inoperable resulting in low process yield rates. In this work, we report a low temperature hybrid assembly approach for the generation of functional thermoplastic nanofluidic devices with high process yield rates (>90%) and with a short total assembly time (16 min). The approach involves thermally sealing a high T(g) (glass transition temperature) substrate containing the nanofluidic structures to a cover plate possessing a lower T(g). Nanofluidic devices with critical feature sizes ranging between 25-250 nm were fabricated in a thermoplastic substrate (T(g) = 104 °C) and sealed with a cover plate (T(g) = 75 °C) at a temperature significantly below the T(g) of the substrate. Results obtained from sealing tests revealed that the integrity of the nanochannels remained intact after assembly and devices were useful for fluorescence imaging at high signal-to-noise ratios. The functionality of the assembled devices was demonstrated by studying the stretching and translocation dynamics of dsDNA in the enclosed thermoplastic nanofluidic channels.


Pharmacokinetics of Transdermal Etofenamate and Diclofenac in Healthy Volunteers.
Little is known about the course of the plasma concentration and the bioavailability of non-steroidal anti-inflammatory drugs (NSAIDs) contained in dermal patches. We compared an etofenamate prototype patch (patent EP 1833471) and a commercially available diclofenac epolamine patch regarding the bioavailability of the active ingredients relative to respective i.m. applications and regarding their plasma concentration-time course. Twenty-four healthy human volunteers were treated using a parallel group design (n = 12 per group) with a single dermal patch (removed after 12 hr) followed (after a latency of 48 hr) by eight consecutive dermal patches every 12 hr to reach steady-state conditions. The patches were generally well tolerated, but one volunteer treated with etofenamate developed an allergic contact dermatitis. After the first patch, Cmax was 0.81 ± 0.11 (mean ± S.E.M.) ng/mL (reached 12 hr after patch removal) for diclofenac and 31.3 ± 3.8 ng/mL for flufenamic acid (reached at patch removal), the main metabolite of etofenamate. Etofenamate was not detectable. After repetitive dosing, trough plasma concentrations after the eighth dose were 1.72 ± 0.32 ng/mL for diclofenac and 48.7 ± 6.6 ng/mL for flufenamic acid. Bioavailabilities (single dose) relative to i.m. applications were 0.22 ± 0.04% for diclofenac and 1.15 ± 0.06% for flufenamic acid. In conclusion, the relative bioavailability (compared to the respective i.m. application) of both drugs is low. The maximal plasma concentrations after topical administration of these drugs are well below the IC50 values for COX-1 and COX-2, explaining the absence of dose-dependent toxicities.


[German version of the Strength and Difficulties Questionnaire (SDQ-German)--overview and evaluation of initial validation and normative results].
The Strengths and Difficulties Questionnaire (SDQ) is a short behavioural screening questionnaire which can be completed in about five minutes by parents and teachers of 4- to 16-year-olds or as self-report by 11- to 16-year-olds. The English original has already been fully evaluated and is widely used in research and clinical practice. The instrument was translated into German in 1997, and several evaluative studies have since been completed. The present paper gives an overview of this novel instrument and summarizes the normative and validation studies to date. A normative study on a field sample of 930 children demonstrated that the distributions of raw scores in the German parent SDQ closely resemble those found in the English version, while a factor analysis of the German data yielded a pattern of loadings which convincingly replicated the original scale structure. Initial validation studies showed that the parent-, teacher-, and self-completed SDQ-Deu correlates well with the considerably longer German versions of the Child Behavior Checklist (CBCL) and respective teacher and self-report derivatives (TRF, YSR). Both parent-rated instruments are equally able to distinguish between a community and a clinic sample, and between subgroups with and without specified categories of disorders within a clinic sample. After discussing possible uses of the SDQ-Deu as well as similarities and differences to other scales, we conclude that the German SDQ is just as useful and valid an instrument for many clinical and research purposes as the English original.


Cholesterol-binding protein TSPO2 coordinates maturation and proliferation of terminally differentiating erythroblasts.
TSPO2 (translocator protein 2) is a transmembrane protein specifically expressed in late erythroblasts and has been postulated to mediate intracellular redistribution of cholesterol. We identified TSPO2 as the causative gene for the HK (high-K+) trait with immature red cell phenotypes in dogs and investigated the effects of the TSPO2 defects on erythropoiesis in HK dogs with the TSPO2 mutation and Tspo2 knockout (Tspo2-/-) mouse models. Bone marrow-derived erythroblasts from HK dogs showed increased binucleated and apoptotic cells at various stages of maturation and shed large nuclei with incomplete condensation when cultured in the presence of erythropoietin, indicating impaired maturation and cytokinesis. The canine TSPO2 induces cholesterol accumulation in the endoplasmic reticulum and could thereby regulate cholesterol availability by changing intracellular cholesterol distribution in erythroblasts. Tspo2-/- mice consistently showed impaired cytokinesis with increased binucleated erythroblasts, resulting in compensated anemia, and their red cell membranes had increased Na,K-ATPase, resembling the HK phenotype in dogs. Tspo2-deficient mouse embryonic stem cell-derived erythroid progenitor (MEDEP) cells exhibited similar morphological defects associated with a cell-cycle arrest at the G2/M phase, resulting in decreased cell proliferation and had a depletion in intracellular unesterified and esterified cholesterol. When the terminal maturation was induced, Tspo2-/- MEDEP cells showed delays in hemoglobinization; maturation-associated phenotypic changes in CD44, CD71, and TER119 expression; and cell-cycle progression. Taken together, these findings imply that TSPO2 is essential for coordination of maturation and proliferation of erythroblasts during normal erythropoiesis.


Listeria monocytogenes ability to survive desiccation: Influence of serotype, origin, virulence, and genotype.
Listeria monocytogenes, a bacterium that is responsible for listeriosis, is a very diverse species. Desiccation resistance has been rarely studied in L. monocytogenes, although it is a stress that is largely encountered by this microorganism in food-processing environments and that could be managed to prevent its presence. The objective of this study was to evaluate the resistance of 30 L. monocytogenes strains to moderate desiccation (75% relative humidity) and evaluate the correlation of such resistance with the strains' virulence, serotype and genotype. The results showed a great heterogeneity of strains regarding their ability to survive (loss of cultivability between 0.4 and 2.0 log). Strains were classified into three groups according to desiccation resistance (sensitive, intermediate, or resistant), and the strain repartition was analyzed relative to serotype, virulence level and environmental origin of the strains. No correlation was found between isolate origin and desiccation resistance. All serotype 1/2b strains were classified into the group of resistant strains. Virulent and hypovirulent strains were distributed among the three groups of desiccation resistance. Finally, a genomic comparison was performed based on 31 genes that were previously identified as being involved in desiccation resistance. The presence of those genes was localized among the genomes of some strains and compared regarding strain-resistance levels. High nucleotide conservation was identified between resistant and desiccation-sensitive strains. In conclusion, the findings regarding the strains of serotype 1/2b indicate potential serotype-specific resistance to desiccation, and thus, to relative humidity fluctuations potentially encountered in food-related environments. The genomic comparison of 31 genes associated to desiccation tolerance did not reveal differences among four strains which have different level of resistance to desiccation.


[Rhizomelic pseudopolyarthritis and Horton's disease. A search for correlations with HLA system, HB virus and Beta 2 microglobulin (author's transl)].
The authors studied 31 patients with either rhizomelic pseudopolyarthritis or Horton's disease. Contrary to others, they found no significant correlation between these diseases and hepatitis B virus infection or HLA gene. However, beta 2 microglobulin assays showed that it was significantly higher in these patients than in a control population. In the absence of close correlation between beta 2 microglobulin levels and those of inflammatory proteins, it is suggested that the rise in microglobulin reflected an immunological mechanism requiring further investigation.


[Mesenterico-caval shunt with interposition of a prosthesis for intractable ascites caused by cirrhosis].
The interposition mesocaval shunt is followed both by a splancnic and sinusoidal decompression, being for this reason considered not only the most effective peripheral porto-systemic shunt, but also one of the indicated operations in the treatment of intractable cyrrhotic ascites. Intractable cyrrhotic ascites was the main indication in 9 cases with very different clinical pictures treated by the Author with mesocaval shunt, having observed no operative mortality and resulting a complete disappearance of ascites in 8 out of 9 cases and of the cohexistent idrothorax in one case. Angiographic pictures showed the patency of the shunt with constant demonstration, after few months, of a reversal portal blood flow (sometimes present before operation). This shunting procedure proved to be a very well tolerated operation with only a transient postoperative modification of liver function tests, excepted for one case in which the patient was lost after seven months because of hepatic encephalopaty with progressive liver failure.


Role of thromboxane in interleukin 2-induced lung injury in sheep.
Interleukin (IL)-2 administration leads to respiratory dysfunction due to increased vascular permeability. This study examines the role of thromboxane (Tx)A2 in IL-2 induced lung injury in sheep with chronic lung lymph fistulae. This preparation enables evaluation of permeability prior to the development of gross edema. IL-2, 10(5) units/kg (n = 6), or its excipient control (n = 5) was given as an i.v. bolus over 2 min. After 2 h of IL-2 administration, plasma TxB2 increased from 168 to 388 pg/ml (P less than 0.05) and lung lymph TxB2 from 235 to 694 pg/ml (P less than 0.05). Mean pulmonary artery pressure (MPAP) rose from 13 to 29 mm of Hg (P less than 0.05) at 30 min and remained elevated for 4 h while the pulmonary artery wedge pressure was unchanged at 4 mm of Hg. Arterial oxygen tension (PaO2) fell from 88 to 77 mm of Hg (P less than 0.05). Lung lymph flow (QL) rose from 2.2 to 3.8 ml/30 min (P less than 0.05) at 1 h and to 6.4 ml/30 min at 3 h. This rise coincided with an increase in the lymph/plasma (L/P) protein ratio from 0.67 to 0.77 (P less than 0.05). In contrast, the non-IL-2-infused sheep (n = 3) recruitment of the lung vasculature by left atrial balloon inflation led to a rise in QL from 2.4 to 8.2 ml/30 min, whereas the L/P ratio declined from 0.62 to 0.25, suggesting that the protein-rich lymph flow after IL-2 administration reflected increased microvascular permeability. In further proof of an increase in permeability, IL-2 administration into sheep (n = 2) with an inflated left atrial balloon led, after a pressure-independent L/P protein ratio had been achieved, to an increase in L/P protein ratio and decrease in protein reflection coefficient. At 2 h after IL-2, the blood leukocyte count fell from 8156 to 4375/mm3 (P less than 0.05) primarily due to a 73% drop in lymphocytes. The platelet count declined from 292 to 184 x 10(3)/mm3 (P less than 0.05). Body temperature rose from 38.9-40.3 degrees C (P less than 0.05), and shaking chills were common. Pretreatment with the Tx synthetase inhibitor OKY 046 (n = 7) lowered baseline plasma and lymph TxB2 levels to 22 and 52 pg/ml (P less than 0.05) and prevented the IL-2-induced increase in plasma and lung lymph TxB2 (P less than 0.05).(ABSTRACT TRUNCATED AT 400 WORDS)


Oct4GiP reporter assay to study genes that regulate mouse embryonic stem cell maintenance and self-renewal.
Pluripotency and self-renewal are two defining characteristics of embryonic stem cells (ES cells). Understanding the underlying molecular mechanism will greatly facilitate the use of ES cells for developmental biology studies, disease modeling, drug discovery, and regenerative medicine (reviewed in (1,2)). To expedite the identification and characterization of novel regulators of ES cell maintenance and self-renewal, we developed a fluorescence reporter-based assay to quantitatively measure the self-renewal status in mouse ES cells using the Oct4GiP cells (3). The Oct4GiP cells express the green fluorescent protein (GFP) under the control of the Oct4 gene promoter region (4,5). Oct4 is required for ES cell self-renewal, and is highly expressed in ES cells and quickly down-regulated during differentiation (6,7). As a result, GFP expression and fluorescence in the reporter cells correlates faithfully with the ES cell identity (5), and fluorescence-activated cell sorting (FACS) analysis can be used to closely monitor the self-renewal status of the cells at the single cell level (3,8). Coupled with RNAi, the Oct4GiP reporter assay can be used to quickly identify and study regulators of ES cell maintenance and self-renewal (3,8). Compared to other methods for assaying self-renewal, it is more convenient, sensitive, quantitative, and of lower cost. It can be carried out in 96- or 384-well plates for large-scale studies such as high-throughput screens or genetic epistasis analysis. Finally, by using other lineage-specific reporter ES cell lines, the assay we describe here can also be modified to study fate specification during ES cell differentiation.


Applying Non-Homogeneous Dose Optimization to Improve Conventionally Fractionated Radiation Plan Quality in Patients with Non-Small Cell Lung Cancer.
Nonhomogeneous dose optimization (NHDO) is exploited in stereotactic body radiation therapy (SBRT) to increase dose delivery to the tumor and allow rapid dose falloff to surrounding normal tissues. We investigate changes in plan quality when NHDO is applied to inverse-planned conventionally fractionated radiation therapy (CF-RT) plans in patients with non-small cell lung cancer. Patients with near-central non-small cell lung cancer treated with CF-RT in 2018 at a single institution were identified. CF-RT plans were replanned using NHDO techniques, including normalizing to a lower isodose line, while maintaining clinically acceptable normal tissue constraints and target coverage. Tumor control probabilities were calculated. We compared delivered CF-RT plans using homogenous dose optimization (HDO) versus NHDO using Wilcoxon signed-rank tests. Median values are reported. Thirteen patients were replanned with NHDO techniques. Planning target volume coverage by the prescription dose was similar (NHDO = 96% vs HDO = 97%, P = .3). All normal-tissue dose constraints were met. NHDO plans were prescribed to a lower-prescription isodose line compared with HDO plans (85% vs 97%, P = .001). NHDO increased mean dose to the planning target volume (73 Gy vs 67 Gy), dose heterogeneity, and dose falloff gradient (P < .03). NHDO decreased mean dose to surrounding lungs, esophagus, and heart (relative reduction of 6%, 14%, and 15%, respectively; P < .05). Other normal tissue objectives improved with NHDO, including total lung V40 and V60, heart V30, and maximum esophageal dose (P < .05). Tumor control probabilities doubled from 31.6% to 65.4% with NHDO (P = .001). In select patients, NHDO principles used in SBRT optimization can be applied to CF-RT. NHDO results in increased tumor dose, reduction in select organ-at-risk dose objectives, and better maintenance of target coverage and normal-tissue constraints compared with HDO. Our data demonstrate that principles of NHDO used in SBRT can also improve plan quality in CF-RT.


Home Health And Postacute Care Use In Medicare Advantage And Traditional Medicare.
This article compares patterns of postacute care-including care provided by skilled nursing facilities, inpatient rehabilitation facilities, and home health agencies-under Medicare Advantage and traditional Medicare. Overall, Medicare Advantage enrollees received less postacute care, both institutional and home health, than traditional Medicare enrollees did for three common conditions.


Garlicnins B(1), C(1), and D, from the fraction regulating macrophage activation of Allium sativum.
Several novel sulfides from acetone extracts of bulbs of garlic (Allium sativum L.), were identified and investigated. These were named garlicnins B(1) (1), C(1) (2), and D (3), and they were found to have the ability to control macrophage activation. Garlicnins B(1) (1) and C(1) (2) possess a new skeleton of cyclic sulfoxide and their structures of garlicnins B(1) (1) and C(1) (2) were characterized as 3,4-dimethyltetrahydrothiophene-S-oxide derivatives carrying the substitutions of a propenyl and a sulfenic acid, and an allyldithiine and a 1-propene-sulfenic acid (a), respectively. The mechanism of the proposed production of these compounds is discussed. Garlicnin D (3), dithiine-type, was estimated to be derived by addition of (a)+allyl thiosulfenic acid (b) derived from allicin. The identification of these novel sufoxides from onion and garlic accumulates a great deal of new chemistry to the Allium sulfide field, and future pharmacological investigations aid the development of natural, healthy foods and anti-cancer agents that could potentially prevent or combat disease.


Twelve Years Implementation of Cervical and Breast Cancer Screening Program in Indonesia.
The study aimed to measure achievement of the national program of cervical and breast cancer screening in Indonesia after 12 years implementation and factors associated with the number of the screening. This was a cross-sectional study with descriptive and analytic analysis. Secondary data was collected from Directorate of Non Communicable Disease Control, Ministry of Health. From 2007 to 2018, the program was implemented in all 34 provinces, at 51% primary health centers (PHC) with 3 providers each. Total women aged 30-50 years screened was 3,664,625 (9.8% of the target). The number rose gradually from 2007 to 2014, with significant increase from 2015 to 2018. Bali province had the highest coverage (31%) and Papua had the lowest (1%). We found a wide disparity of coverage among provinces. There was 3.4% of VIA-positive, 16.1% was treated with cryotherapy, 1.3 per 1,000 of suspected cervical cancer, 5,4% lump in the breast, and 0.7 per 1,000 suspected breast cancer. Factors associated with number of the screening were number of PHC providing screening, number of GP, total provider, number of NCD post, number of Village with NCD Post, and income of the province. The cervical and breast cancer screening program was running in all provinces in more than half of primary health centers in Indonesia. National coverage (9.8%) was far below the target and varied widely among provinces. Number of PHC with screening services, number of GP, number of total provider, number of NCD post, number of Village with NCD Post, and income of the province have association with cervical and breast cancer screening.


Distribution of the cyanophyte Trichodesmium (Oscillatoriaceae) in the eastern Caribbean Sea: influence of the Orinoco River.
Orinoco River influence in the Caribbean Sea, characterized by high nutrient input, causes a decrease of Trichodesmium populations. The Caribbean Time Series (CaTS) station, south of Puerto Rico (17 degrees 36'N 67 degrees 00'W), was monitored for 25 months in order to observe the Trichodesmium abundance pattern and the presence of the river plume. In general, mean Trichodesmium abundance was higher at the surface and decreased with depth. The mean upper water column (surface to 20 m) abundance was 54.1 +/- 32.6 col/m3. Within the sampling period, abundance was highly variable (1-700 col/m3). Correlation between Trichodesmium abundance and wind speed (p=0.002), chlorophyll a concentration (p=0.001), nitrate (p=0.02) and silicate (p=0.003) concentrations were statistically significant. However, Trichodesmium abundance was not correlated with salinity (p=0.70), temperature (p=0.16) and seawater density (p=0.71) variations at CaTS. Eastern Caribbean regions highly influenced by the Orinoco River discharge were devoid of Trichodesmium colonies.


Health-care utilization and associated factors in Gauteng province, South Africa.
More than a billion people, mainly in low- and middle-income countries, are unable to access needed health-care services for a variety of reasons. Possible factors influencing health-care utilization include socio-demographic and economic factors such as age, sex, education, employment and income. However, different studies have showed mixed results. Moreover, there are limited studies on health-care utilization. This study aimed to determine health-care utilization and associated factors among all residents aged 18 or over in Gauteng province, South Africa. A cross-sectional study was conducted from data collected for a Quality of Life survey which was carried out by Gauteng City-Region Observatory in 2013. Simple random sampling was used to select participants. A total of 27,490 participants have been interviewed. Data were collected via a digital data collection instrument using an open source system called Formhub. Coarsened Exact Matching (CEM) was used to improve estimation of causal effects. Stepwise multiple logistic regression was employed to identify factors associated with health-care utilization. Around 95.7% reported usually utilizing health-care services while the other 4.3% reported not having sought health-care services of any type. Around 75% of participants reported reduced quality of public health services as a major reason not to visit them. Higher odds of reported health-care utilization were associated with being female (OR = 2.18, 95% CI: 1.88-2.53; p < 0.001), being White compared to being African (OR = 2.28, 95% CI: 1.84-2.74; p < 0.001), and having medical insurance (OR = 5.41, 95% CI: 4.06-7.23; p < 0.001). Lower odds of seeking health-care were associated with being an immigrant (OR = 0.61, 95% CI: 0.53-0.70; p < 0.001). The results indicated that there is a need to improve the quality of public health-care services and perception towards them as improved health-care quality increases the choice of health-care providers.


A Case of Tubular Adenoma of Gallbladder Diagnosed Using Contrast-Enhanced Ultrasonography.
Contrast-enhanced ultrasound (CEUS) has been used to diagnose gallbladder (GB) diseases for recent years because it is sensitive to visualize vascularity. Herein, we report a case who had a 1.7 cm × 1.2 cm polypoid lesion located in the gallbladder fundus with a feeding artery located in the stalk. On CEUS, the lesion showed early arterial phase enhancement (time to peak enhancement 18 s), persisting throughout the venous and delay phases. This enhancing pattern suggested that the lesion was a GB adenoma rather than a GB carcinoma. Cholecystectomy was performed, and pathology of the tissue revealed tubular adenoma of the GB.


Serum stimulation of phospholipase A2 and prostaglandin release in 3T3 cells is associated with platelet-derived growth-promoting activity.
Sera from mouse, rat, and calf sources stimulate cellular phospholipase A2 activity (PLase; phosphatide 2-acylhydrolase, EC 184.108.40.206) and prostaglandin synthesis in 3T3 Swiss mouse fibroblasts, releasing up to 33% of biosynthetically incorporated [3H]arachidonic acid as hydrolysis products in 1 hr. The PLase stimulated by mouse serum exhibits specificity for arachidonic acid residues on phospholipids. It is stimulated 2.5-fold by 1.8 mM Ca2+ in the presence of 5 microM divalent cation ionophore A23187, consistent with a Ca2+-dependent enzyme possessing a cytoplasmic Ca2+ binding site. The percentage maximal PLase- and growth-stimulating activities of the three sera exhibit similar concentration dependencies, with the homologous (mouse) serum exhibiting the highest specific activities. Confluent 3T3 cells deplete PLase-stimulating activity from medium containing calf serum at a rate similar to the depletion of cell growth-stimulus activity. The PLase-stimulating activity in rat and mouse sera is derived from the leukocyte fraction of blood, presumably from platelets. In rat leukocyte lysates the PLase-stimulating activity exhibits properties similar to those reported for platelet-derived cell growth factors from rat and human sources--i.e., stability to exposure to 100 degrees C for 2 min or to pH 2 or pH 11, and cationic properties. Purified preparations of human platelet-derived growth factor also exhibit 3T3 PLase-stimulating activity.


Decreased platelet function in Cavalier King Charles Spaniels with mitral valve regurgitation.
With aggregometry, increased platelet activity has been reported in Cavalier King Charles Spaniels (CKCS) without mitral regurgitation (MR). In contrast, dogs with MR have been found to have decreased platelet activity. The purpose of this study was to test an easy bedside test of platelet function (the Platelet Function Analyzer [PFA-100]) to see if it could detect an increase in platelet activity in CKCS without MR and a decrease in platelet activity in CKCS with MR. This study included 101 clinically healthy dogs > 1 year of age: 15 control dogs of different breeds and 86 CKCS. None of the dogs received medication or had a history of bleeding. The PFA-100 evaluates platelet function in anticoagulated whole blood under high shear stress. Results are given as closure times (CT): the time it takes before a platelet plug occludes a hole in a membrane coated by agonists. The CT with collagen and adenosine-diphosphate as agonists was similar in control dogs (median 62 seconds; interquartile interval 55-66 seconds) and CKCS with no or minimal MR (55; 52-64 seconds). The CT was higher in CKCS with mild MR (regurgitant jet occupying 15-50% of the left atrial area) (75; 60-84 seconds; P = .0007) and in CKCS with moderate to severe MR (jet > 50%) (87: 66-102 seconds; P < .0001). CKCS with mild, moderate, and severe, clinically inapparent MR have decreased platelet function. The previous finding of increased platelet reactivity in nonthrombocytopenic CKCS without MR could not be reproduced with the PFA-100 device.


Up-regulation of transferrin receptor gene expression by granulocyte colony-stimulating factor in human myeloid leukemia cells.
Granulocyte colony-stimulating factor (G-CSF) enhanced surface transferrin receptor (TfR) expression in two human myeloid leukemia cell lines, NKM-1 and NOMO-1, which possess G-CSF receptors. Radioligand-binding assay revealed that 10 ng/ml G-CSF significantly increased TfR to 186 +/- 20 and 276 +/- 38% of control for NKM-1 cells and NOMO-1 cells, respectively, in a 24-h culture. Scatchard analysis showed the increase of transferrin (Tf)-binding sites but no change in the receptor affinity. The enhanced TfR expression was not mediated either by the kinetic change of receptor cycling or by cellular iron content. Immunoprecipitation with anti-TfR antibody was used, and the increased biosynthesis of the receptor was demonstrated in G-CSF-stimulated cells. Northern blot analysis showed a 2- to 3-fold increase of TfR mRNA of NKM-1 cells cultured in medium containing Tf and G-CSF, whereas the mRNA declined without G-CSF. The effect of G-CSF on the TfR mRNA was observed within 2 h, which preceded the increase of surface TfR and the transition to the S phase of the cell cycle. G-CSF also potentiated TfR expression in freshly obtained myeloid leukemia cells. The present study shows up-regulation of TfR expression by G-CSF in myeloid leukemia cells and provides evidence that the regulation is mediated by controlling the steady-state level of the mRNA.


Functional Neuroimaging Evidence for Distinct Neurobiological Pathways in Attention-Deficit/Hyperactivity Disorder.
A challenge facing clinical neuroscientists is how best to synthesize diverse and sometimes inconsistent evidence for neuropsychological deficits and brain system dysfunction found in psychiatric disorders into models that guide etiological and treatment research. Multiple-pathway models suggest that psychiatric symptoms might arise from pathophysiology in different neural systems. This study tested dual-pathway model predictions for attention-deficit/hyperactivity disorder (ADHD) that reward and executive function cognitive deficits should be related to abnormalities in corresponding functionally specialized neural systems. Behavioral inhibition and preference for immediate rewards were assessed in N = 251 adolescent boys and girls ages 12 to 18 diagnosed with DSM-IV combined-subtype ADHD or non-ADHD control subjects. Following taxometric analyses of test performance, the resulting subgroups were compared on a functional magnetic resonance imaging monetary incentive delay task probing reward anticipation and go/no-go task of motor response inhibition. Three ADHD subgroups were identified consistent with different proposed pathways-ADHD with executive function/motor inhibition deficits, ADHD with both executive and reward deficits, and ADHD with relatively normal test performance. Each cognitive domain mapped to different ADHD brain dysfunction features as expected. However, no brain abnormalities were found common to all ADHD subgroups despite the fact they had nearly identical ADHD-related clinical characteristics. The results suggest that combined-subtype ADHD is a collection of discrete disorders for which a comparable behavioral end point arises through different neurobiological pathways. The findings raise caution about applying common cause, single-deficit conceptual models to individual ADHD patients and should prompt researchers to consider biologically defined, multifactorial etiological models for other psychiatric diagnoses.


Combined All-Extremity High-Intensity Interval Training Regulates Immunometabolic Responses through Toll-Like Receptor 4 Adaptors and A20 Downregulation in Obese Young Females.
Metainflammation and malfunctions of toll-like receptor 4 (TLR4) are related to obesity-induced immunometabolic morbidities. There are almost no studies relating exercise training to the TLR4 pathway and its adaptors and negative regulators. Thirty young women with obesity (exercise group and control group) were included in a 10-week all-extremity combined high-intensity interval training program. The immunomodulatory impacts of exercise on TLR4, its related adaptors (TIR domain-containing adaptor-inducing IFN-β[TRIF], myeloid differentiation factor 88 [MyD88],and tumor receptor-associated factor 6 [TRAF6]), transcriptional factors (nuclear factor [NF]-κB and interferon regulatory factor 3 [IRF3]), and negative regulator (A20) mRNA levels were assessed by real-time PCR. Also, the serum concentration of TLR4 final products (tumor necrosis factor α [TNFα] and interferon γ [IFNγ]) was measured by ELISA. Cardiorespiratory and body composition parameters were tested, as well. There was a significant improvement in body composition and cardiorespiratory fitness. This intervention downregulated TLR4 (from 2.25 ± 1.07 to 0.84 ± 1.01), MyD88 (from 4.53 ± 5.15 to 1.27 ± 0.88), NF-κB (from 1.61 ± 2.03 to 0.23 ± 0.39), IRF3 (from 1.22 ± 0.77 to 0.25 ± 0.36), and A20 (from 0.88 ± 0.59 to 0.22 ± 0.33) levels and reduced the TNFα concentrations (from 22.39 ± 11.43 to 6.26 ± 5.31) significantly in the exercise group, while no statistically significant change was found in TRIF and TRAF6 expression and IFNγ circulating levels. It is concluded that long-term exercise modifies the inflammatory pathways and modulates the immune function at the early stages of inflammation initiation in circulating immune cells. Accordingly, we suggest time-efficient exercise protocols as a possible therapy approach for the prevention of M1 polarization.


Gel-forming mucins appeared early in metazoan evolution.
Mucins are proteins that cover and protect epithelial cells and are characterized by domains rich in proline, threonine, and serine that are heavily glycosylated (PTS or mucin domains). Because of their sequence polymorphism, these domains cannot be used for evolutionary analysis. Instead, we have made use of the von Willebrand D (VWD) and SEA domains, typical for mucins. A number of animal genomes were examined for these domains to identify mucin homologues, and domains of the resulting proteins were used in phylogenetic studies. The frog Xenopus tropicalis stands out because the number of gel-forming mucins has markedly increased to at least 25 as compared with 5 for higher animals. Furthermore, the frog Muc2 homologues contain unique PTS domains where cysteines are abundant. This animal also has a unique family of secreted mucin-like proteins with alternating PTS and SEA domains, a type of protein also identified in the fishes. The evolution of the Muc4 mucin seems to have occurred by recruitment of a PTS domain to AMOP, NIDO, and VWD domains from a sushi domain-containing family of proteins present in lower animals, and Xenopus is the most deeply branching animal where a protein similar to the mammalian Muc4 was identified. All transmembrane mucins seem to have appeared in the vertebrate lineage, and the MUC1 mucin is restricted to mammals. In contrast, proteins with properties of the gel-forming mucins were identified also in the starlet sea anemone Nematostella vectensis, demonstrating an early origin of this group of mucins.


ATP-dependent minor groove recognition of TA base pairs is required for template melting by the E1 initiator protein.
Template melting is an essential step in the initiation of DNA replication, but the mechanism of template melting is unknown for any replicon. Here we demonstrate that melting of the bovine papillomavirus type 1 ori is a sequence-dependent process which relies on specific recognition of TA base pairs in the minor groove by the E1 initiator. We show that correct template melting is a prerequisite for the formation of a stable double hexamer with helicase activity and that ori mutants that fail to melt correctly are defective for ori unwinding and DNA replication in vivo. Our results also indicate that melting of the DNA is achieved by destabilization of the double helix along its length through multiple interactions with E1, each of which is responsible for melting of a few base pairs, resulting in the extensive melting that is required for initiation of DNA replication.


Transcriptome analysis during berry development provides insights into co-regulated and altered gene expression between a seeded wine grape variety and its seedless somatic variant.
Seedless grapes are greatly appreciated for fresh and dry fruit consumption. Parthenocarpy and stenospermocarpy have been described as the main phenomena responsible for seedlessness in Vitis vinifera. However, the key genes underpinning molecular and cellular processes that play a significant role in seed development are not well characterized. To identify important regulators and mechanisms that may be altered in the seedless phenotype, we performed a comprehensive transcriptional analysis to compare the transcriptomes of a popular seeded wine cultivar (wild-type) and its seedless somatic variant (mutant) at three key developmental stages. The transcriptomes revealed by Illumina mRNA-Seq technology had approximately 98% of grapevine annotated transcripts and about 80% of them were commonly expressed in the two lines. Differential gene expression analysis revealed a total of 1075 differentially expressed genes (DE) in the pairwise comparison of developmental stages, which included DE genes specific to the wild-type background, DE genes specific to the mutant background and DE genes commonly shared in both backgrounds. The analysis of differential expression patterns and functional category enrichment of wild-type and mutant DE genes highlighted significant coordination and enrichment of pollen and ovule developmental pathways. The expression of some selected DE genes was further confirmed by real-time RT-PCR analysis. This study represents the most comprehensive attempt to characterize the genetic bases of seed formation in grapevine. With a high throughput method, we have shown that a seeded wine grape and its seedless somatic variant are similar in several biological processes. Nevertheless, we could identify an inventory of genes with altered expression in the mutant compared to the wild-type, which may be responsible for the seedless phenotype. The genes located within known genomic regions regulating seed content may be used for the development of molecular tools to assist table grape breeding. Therefore the data reported here have provided a rich genomic resource for practical use and functional characterization of the genes that potentially underpin seedlessness in grapevine.


Ag-Mg antisite defect induced high thermoelectric performance of α-MgAgSb.
Engineering atomic-scale native point defects has become an attractive strategy to improve the performance of thermoelectric materials. Here, we theoretically predict that Ag-Mg antisite defects as shallow acceptors can be more stable than other intrinsic defects under Mg-poor‒Ag/Sb-rich conditions. Under more Mg-rich conditions, Ag vacancy dominates the intrinsic defects. The p-type conduction behavior of experimentally synthesized α-MgAgSb mainly comes from Ag vacancies and Ag antisites (Ag on Mg sites), which act as shallow acceptors. Ag-Mg antisite defects significantly increase the thermoelectric performance of α-MgAgSb by increasing the number of band valleys near the Fermi level. For Li-doped α-MgAgSb, under more Mg-rich conditions, Li will substitute on Ag sites rather than on Mg sites and may achieve high thermoelectric performance. A secondary valence band is revealed in α-MgAgSb with 14 conducting carrier pockets.


1D Genome Sequencing on the Oxford Nanopore MinION.
Today's short-read sequencing instruments can generate read lengths between 50 bp and 700 bp depending on the specific instrument. These high-throughput sequencing approaches have revolutionized genomic science, allowing hundreds of thousands of full genomes to be sequenced, and have become indispensable tools for many researchers. With greater insight has come the revelation that many genomes are much more complicated than originally thought and include many rearrangements and copy-number variations. Unfortunately, short-read sequencing technologies are not well suited for identifying many of these types of events. Long-read sequencing technologies can read contiguous fragments of DNA in excess of 10 kb and are much better suited for detecting large structural events. The newest long-read sequencing instrument is the MinION device from Oxford Nanopore. The rapid sequencing speed and low upfront instrument cost are features drawing interest in this device from the genomics community. This unit provides a representative protocol for carrying out human genome sequencing on the Oxford Nanopore MinION. © 2017 by John Wiley & Sons, Inc.


Onset of slow dynamics in difluorotetrachloroethane glassy crystal.
Complementary neutron spin-echo and x-ray experiments and molecular-dynamics simulations have been performed on difluorotetrachloroethane (CFCl2-CFCl2) glassy crystal. Static, single-molecule reorientational dynamics and collective dynamics properties are investigated. Our results confirm the strong analogy between molecular liquids and plastic crystals. The orientational disorder is characterized at different temperatures and a change in the nature of rotational dynamics is observed. A careful check of the rotational diffusion model is performed using self-angular correlation functions Cl with high l values and compared to results obtained on molecular liquids composed of A-B dumbbells. Below the crossover temperature at which slow dynamics emerge, we show that some scaling predictions of the mode coupling theory hold and that alpha-relaxation times and nonergodicity parameters are controlled by the nontrivial static correlations.


Modulation of regulatory T cells by intranasal allergen immunotherapy in an experimental rat model of airway allergy.
Allergic airway diseases such as asthma and allergic rhinitis are increasing in prevalence worldwide. The theory of an altered Th1/Th2 balance in allergic diathesis has recently been termed a "procrustean paradigm" as it failed to explain many preclinical findings. Regulatory T cells (Treg) have now been shown to be critical in T-cell homeostasis and in the maintenance of peripheral tolerance to allergens. Allergen specific immunotherapy (SIT) has been shown to induce regulatory T cells in allergic patients. Among various types of SIT, intranasal immunotherapy had not been studied in detail for the treatment of allergic airway diseases. So, there was a need to study the contribution of regulatory T cells and their mechanistic pathways following intranasal immunotherapy in-vivo. It had been previously shown that intranasal allergen immunotherapy using Alstonia scholaris pollen extract abrogates allergic airway inflammation with decline in IgE and Th2 cytokine levels. The present study for the first time offers a multi-targeted approach towards attenuation of airway allergy by the generation of CD4+CD25+Foxp3+T cells and other subsets of Treg cells like Tr1 cells, Th3 cells, CTLA4+Treg cells, and also modulation of various Treg cell surface molecules like GITR, OX40, CD39 and CD73 by intranasal immunotherapy in the same animal model. This animal experiment will thus help to chart out newer molecular targets for treating allergic asthma or rhinitis.


Progression or resolution of coxsackievirus B4-induced pancreatitis: a genomic analysis.
Group B coxsackieviruses are associated with chronic inflammatory diseases of the pancreas, heart, and central nervous system. Chronic pancreatitis, which can develop from acute pancreatitis, is considered a premalignant disorder because it is a major risk factor for pancreatic cancer. To explore the genetic events underlying the progression of acute to chronic disease, a comparative analysis of global gene expression during coxsackievirus B4-induced acute and chronic pancreatitis was undertaken. A key feature of acute pancreatitis that resolved was tissue regeneration, which was accompanied by increased expression of genes involved in cell growth, inhibition of apoptosis, and embryogenesis and by increased division of acinar cells. Acute pancreatitis that progressed to chronic pancreatitis was characterized by lack of tissue repair, and the expression map highlighted genes involved in apoptosis, acinoductular metaplasia, remodeling of the extracellular matrix, and fibrosis. Furthermore, immune responses appeared skewed toward development of alternatively activated (M2) macrophages and T helper 2 (Th2) cells during disease that resolved and toward classically activated (M1) macrophages and Th1 cells during disease that progressed. Our hypothesis is that growth and differentiation signals coupled with the M2/Th2 milieu favor acinar cell proliferation, while diminished growth signals and the M1/Th1 milieu favor apoptosis of acinar cells and remodeling/proliferation of the extracellular matrix, resulting in fibrosis.


Construction of a confidence set of markers for the location of a disease gene using affected sib pair data.
We have previously proposed a confidence set approach for finding tightly linked genomic regions under the setting of parametric linkage analysis. In this article, we extend the confidence set approach to nonparametric linkage analysis of affected sib pair (ASP) data based on their identity-by-descent (IBD) information. Two well-known statistics in nonparametric linkage analysis, the Two-IBD test (proportion of ASPs sharing two alleles IBD), and the Mean test (average number of alleles shared IBD in the ASPs), are used for constructing confidence sets. Some numerical analyses as well as a simulation study were carried out to demonstrate the utility of the methods. Our results show that the fundamental advantages of the confidence set approach in parametric linkage analysis are retained when the method is generalized to nonparametric analysis. Our study on the accuracy of confidence sets, in terms of choice of tests, underlying disease incidence data, and amount of data available, leads us to conclude, among other things, that the Mean test outperforms the Two-IBD test in most situations, with the reverse being true only for traits with small additive variance. Although we describe how to construct confidence sets based on only two familiar tests, one can construct confidence sets similarly using other allele sharing statistics.


Analysis of corneal topography after excimer laser photorefractive keratectomy.
Excimer laser photorefractive keratectomy (PRK) is an effective treatment for myopia. We investigated the magnitude of optical zone decentration and qualitative patterns of corneal topography after this treatment. We performed computer-assisted videokeratography on 44 myopic eyes (29 patients) that had undergone PRK before and 1, 6, and 12 months after the procedure. Associations of clinical outcomes with decentration and topographic patterns were assessed. The normalized scale of the topography 1 month postoperatively showed a mean decentration of 0.33 +/- 0.23 mm (range 0-0.9). Thirty-four eyes had decentration of less than 0.50 mm; 10 had an ablation zone decentered from 0.5 to 0.9 mm. Analysis of geometric mean visual acuities between eyes with less than 0.5 mm decentration and those with 0.5 to 0.9 mm decentration demonstrated minimal differences. No eye was decentered more than 1 mm. Four main ablation patterns were noted on subtraction analysis: homogeneous, semicircular, keyhole, and central island. Over time, the number of eyes with a homogeneous pattern increased. Eyes with a homogeneous ablation pattern had significantly better uncorrected visual acuity than those with other patterns. The mean visual acuity was 20/29.1 in the homogeneous group and 20/38.5 in the pooled irregular group 1 month postoperatively (p < 0.05). There was no significant difference among the four ablation patterns at 6 or 12 months after PRK. Topographic patterns were not significantly associated with best-corrected vision.


Managing inhaled anesthesia: challenges from a health-system pharmacist's perspective.
To discuss the challenges that health-system pharmacists face in managing the use and costs of inhaled anesthesia in the operating room (OR), the characteristics of inhaled anesthesia agents, systems for delivering inhaled anesthesia, and the role of inhaled anesthetics in fast-track recovery after surgery. Inhaled anesthetic agents are the most common drugs used in general anesthesia and are a substantial part of the anesthetic drug budget in health systems. Challenges for health-system pharmacists in managing costs associated with these agents include the lack of a dedicated OR pharmacy, limited access to the OR, unfamiliarity with some of the medications used in the OR, and difficulty quantifying inhaled anesthetic drug use. The three inhaled anesthesia agents currently used in the United States have proven to be safe and effective. These agents have differences in solubility in blood and tissues, which affect onset, absorption, and excretion. Isoflurane has the highest solubility in blood and tissues, which may result in slower recovery. Lower solubility allows for a faster recovery. The two newest agents on the market, sevoflurane and desflurane, both have low solubility, with desflurane having lower solubility than sevoflurane. Sevoflurane has the advantage of low pungency and is not associated with respiratory irritation. As a cost-savings initiative, the inhalation agents may be used with low flow rates, which minimizes the amount of inhaled anesthetic used. In addition, using the less soluble inhaled anesthetic agents as part of a fast-track approach will accelerate recovery by reducing time to emergence and recovery. This approach can potentially reduce costs to the institution. Understanding the differences in characteristics and delivery of the inhaled anesthesia agents will enable health-system pharmacists to collaborate with anesthesia care providers to better manage the use and costs of these agents.


Delayed hypersensitivity in patients treated by curative radiotherapy. Its relation to tumor response and short-term survival.
Delayed hypersensitivity, as determined by skin reactivity to 2-4 dinitrochlorobenzene (DNCB), was tested in 112 patients prior to their undergoing curative radiotherapy. Of the strong DNCB reactors, 84% had an excellent radiation tumor response, while only 48% of the negative reactors had an equivalent response. Almost twice as many strong DNCB reactors were alive and free of disease (NED) at 6 months (70% vs. 38% for nonreactors). Similarly, the positive DNCB reactors had a median survival of 18 months, compared to 10 months for the nonreactors.


[The importance of HPV vaccination in men].
The important goal of immunization programs in many countries is the reduction of the incidence of cervical cancer using either the quadrivalent (Silgard/Gardasil) or the bivalent (Cervarix) HPV (human papillomavirus) vaccine. Nevertheless, HPV infection is associated with the development of cancers of anus, vagina, vulva and penis, and cancers of the head and neck and genital warts, too. Large trials for both vaccines find efficacy against HPV-related infection and different HPV associated diseases.Infection with HPV and diseases caused by HPV are common in boys and men, too. Approximately 5.2 % of all cancers are HPV associated and the burden of HPV associated disease in men is now comparable to that in women in economically developed countries. Randomized control trials demonstrate robust antibody responses and high efficacy also in men. Several countries recommend gender-neutral vaccination.Detailed cost effective modeling has preceded these decisions showing that when the burden of disease in men is included in the models then, depending upon vaccine price, coverage of a vaccinated population, and other factors male vaccination can become cost effective. Vaccine price had a decisive impact on results. However, increasing coverage in girls is substantially more effective and cost-effective than expanding vaccination coverage to boys and should be considered a priority. Since 2012, vaccination of girls at the age of 13-14 years has been covered from the health insurance in the Czech Republic.


Increased expression levels of Syntaxin 1A and Synaptobrevin 2/Vesicle-Associated Membrane Protein-2 are associated with the progression of bladder cancer.
Gene expression is tightly regulated in time and space through a multitude of factors consisting of signaling molecules. Soluble N-ethylmaleimide-sensitive-factor attachment protein receptors (SNARE) are membrane proteins responsible for the intercellular trafficking of signals through endocytosis and exocytosis of vesicles. Altered expression of SNARE proteins in cellular communication is the major hallmark of cancer phenotypes as indicated in recent studies. SNAREs play an important role in maintaining cell growth and epithelial membrane permeability of the bladder and are not only involved in cancer progression but also metastatic cell invasion through SNARE-mediated trafficking. Synaptobrevin2/Vesicle associated membrane protein-2 (v-SNARE) and Syntaxin (t-SNARE) form a vesicular docking complex during endocytosis. Some earlier studies have shown a critical role of SNARE in colon, lungs, and breast cancer progression and metastasis. In this study, we analyzed the relative expression of the STX1A and VAMP2 (SYB2) for their possible association in the progression and metastasis of bladder cancer. The profiling of the genes showed a significant increase in STX1A and VAMP2 expression (p < 0.001) in high-grade tumor cells compared to normal and low-grade tumors. These findings suggest that elevated expression of STX1A and VAMP2 might have caused the abnormal progression and invasion of cancer cells leading to the transformation of cells into high-grade tumor in bladder cancer.


Temporal requirement of dystroglycan glycosylation during brain development and rescue of severe cortical dysplasia via gene delivery in the fetal stage.
Congenital muscular dystrophies (CMDs) are characterized by progressive weakness and degeneration of skeletal muscle. In several forms of CMD, abnormal glycosylation of α-dystroglycan (α-DG) results in conditions collectively known as dystroglycanopathies, which are associated with central nervous system involvement. We recently demonstrated that fukutin, the gene responsible for Fukuyama congenital muscular dystrophy, encodes the ribitol-phosphate transferase essential for dystroglycan function. Brain pathology in patients with dystroglycanopathy typically includes cobblestone lissencephaly, mental retardation, and refractory epilepsy; however, some patients exhibit average intelligence, with few or almost no structural defects. Currently, there is no effective treatment for dystroglycanopathy, and the mechanisms underlying the generation of this broad clinical spectrum remain unknown. Here, we analysed four distinct mouse models of dystroglycanopathy: two brain-selective fukutin conditional knockout strains (neuronal stem cell-selective Nestin-fukutin-cKO and forebrain-selective Emx1-fukutin-cKO), a FukutinHp strain with the founder retrotransposal insertion in the fukutin gene, and a spontaneous Large-mutant Largemyd strain. These models exhibit variations in the severity of brain pathology, replicating the clinical heterogeneity of dystroglycanopathy. Immunofluorescence analysis of the developing cortex suggested that residual glycosylation of α-DG at embryonic day 13.5 (E13.5), when cortical dysplasia is not yet apparent, may contribute to subsequent phenotypic heterogeneity. Surprisingly, delivery of fukutin or Large into the brains of mice at E12.5 prevented severe brain malformation in Emx1-fukutin-cKO and Largemyd/myd mice, respectively. These findings indicate that spatiotemporal persistence of functionally glycosylated α-DG may be crucial for brain development and modulation of glycosylation during the fetal stage could be a potential therapeutic strategy for dystroglycanopathy.


High-dose steroid-related osteonecrosis in a four-year-old child with acute lymphoblastic leukemia.
Osteonecrosis is an uncommon complication of acute lymphoblastic leukemia. One of the risk factor is high-dose corticosteroid therapy. The highest incidence of osteonecrosis is observed in children 9 to 18 years old at diagnosis and it is a rare condition below 5 years of age. We describe a 4 year-old child with acute lymphoblastic leukemia and complaints of progressive bone pain and walking difficulty who developed osteonecrosis and bone fracture after two remission induction chemotherapy.


Impact of splenectomy on circulating microparticles in patients with sickle cell anemia.
Circulating microparticles (MP) are being described as potential biomarkers for disease activity in a variety of conditions including sickle cell anemia (SCA). However, relatively little is known about the influence of spleen status on MP levels in patients with SCA. Using a prospective study design we characterize circulating MP in 144 patients with SCA in steady state by assessing their cellular origin and their relationships to spleen status defined by clinical and imaging findings. In addition, MP levels were studied according to demographic characteristics, clinical status, treatment modalities, and other hematological and biochemical parameters. Absolute plasma concentrations of MP were determined by flow cytometry. Patients with SCA displayed a 10-fold increase in levels of MP derived from red blood cell (RBC) and platelets (PLT) when compared to their healthy counterparts (p < 0.0001). Splenectomized patients with SCA have more pronounced levels of MPRBC and MPPLT, and remained elevated after several weeks of follow-up. Levels of MP were not significantly associated with spleen removal procedures, age, gender, clinical severity score, hydroxyurea therapy, hemoglobin F, and co-existence of glucose-6-phosphate dehydrogenase deficiency. Collectively, these results suggest that splenectomy affects circulating levels of MP regardless of the known SCA modifiers and correlates.


Chemotherapy Reverses Anti-PD-1 Resistance in One Patient With Advanced Non-small Lung Cell Cancer.
Background: Programmed cell death protein 1(PD-1) blockade has become a standard second-line treatment option for patients with advanced non-small cell lung cancer (NSCLC) without a driver gene mutation. Previous clinical studies showed that the objective response rate (ORR) of PD-1 blockade as second-line treatment for patients with NSCLC was ~20%, and the median progression-free survival (PFS) was ~4 months, with most patients eventually developing a resistance to PD-1 blockade. Although the ORR to chemotherapy after PD-1 blockade resistance was relatively high, the survival time of patients could not be significantly prolonged. Clinical oncologists are unclear about which treatment regimen should be selected after PD-1 blockade failure. Here, we report about a patient with advanced NSCLC and initial PD-1 blockade resistance who was observed to have a rapid partial response (PR) following one dose of chemotherapy and subsequent PD-1 blockade treatment. Case presentation: A 70-year-old woman with a history of left lower lobe lung surgery in March 2018 (pathological stage T1N2M0, EGFR wild-type) presented to our hospital. After six cycles of adjuvant chemotherapy, multiple nodules in both the lungs developed, and were suspected to be metastatic lesions. After another 2 months, the nodules in both the lungs enlarged. From November 2018 to March 2019, the patient received six cycles of pembrolizumab, and computed tomography (CT) confirmed a progressive disease status. She was then managed with 260 mg/m2 albumin paclitaxel once every 3 weeks. Subsequently, chemotherapy was discontinued after one cycle owing to grade three neuromuscular toxicity. Follow-up CT revealed a stable disease in May 2019. She then received another six cycles of pembrolizumab, which resulted in a PR. Conclusion: Chemotherapy may play a role in reversing PD-1 blockade resistance. If failure of PD-1 blockade occurs at first, re-administration of PD-1 blockade may be implemented if first followed by several cycles of chemotherapy. Because there are few reports on the use of chemotherapy to reverse PD-1 resistance, it is necessary to conduct clinical studies with larger patient cohorts to investigate whether chemotherapy can reverse PD-1 blockade resistance.


[A histological and immunohistological study of vascular and inflammatory changes in rosacea].
Rosacea has a number of pathophysiological components, chief of which are vascular abnormalities and inflammation. The morphology of the dilated vessels in rosacea may indicate an increase in the number and size of lymphatic vessels. We carried out a histological and an immunohistological study to quantify these abnormalities in rosacea and compared them with those seen in lupus erythematosus. We reviewed all cases of rosacea analysed over a 4-year period. Ultimately, we only included 86 cases in which the diagnosis could be confirmed by a dermatologist based upon histopathological correlation and follow-up. All biopsies were reviewed for histopathological features, and 25 of these were compared with 25 facial biopsies in documented cases of lupus erythematosus, using standard staining followed by immunohistochemical analysis with anti-CD3, CD4, CD8 and CD20 (lymphocytic) antibodies, anti-CD68 (histiocytic) antibodies, anti-CD31 (endothelial cell) antibodies and anti-D2-40 (podoplanin, a marker for lymphatic endothelial cells) antibodies. In 88% of cases of rosacea, large superficial dermal vessels of geometrical or bizarre configuration were noted, and turgescent cells and dermal edema were frequently seen. Over 75% of cases involved Demodex, including erythemato-telangiectatic subtypes. The rosacea included a mean 15 vessels/mm(2), eight of which expressed D2-40; six were greater than 30μm in diameter (mean: 103μm; maximum: 400μm), with only two of these being D2-40+. The lupus erythematosus biopsies exhibited a mean 15 vessels/mm(2), nine of which expressed D2-40; four measured over 30μm in diameter (mean: 59μm; maximum: 100μm), of which two were D2-40+. The vessels measuring over 100μm were only seen in rosacea, and notable actinic elastosis was associated in 80% of these cases. No Demodex was seen in the lupus cases. The lymphocytic infiltration consisted mainly of CD4+ T cells in both groups, but was chiefly sub-epidermal in lupus, occasionally masking the small vessels of the superficial dermis. Rosacea is characterised by large, dilated, anfractuous capillaries, which are both larger and more numerous than in lupus, although there is no difference in dermal vascular density between the two diseases. Contrary to what their form may suggest, these dilated vessels are not lymphatic. D2-40+ vessels (lymphatic), which are flatter, are found in both lupus and rosacea. The association of large telangiectasias with actinic elastosis may indicate a causative role of exposure to UV radiation. These vessels likely exhibit increased permeability, resulting in dermal edema. Inflammation is consistently present, even in the early forms, strongly suggesting a dual inflammatory and vascular mechanism.


Mucosal healing in the era of biologic agents in treatment of inflammatory bowel disease.
Mucosal healing has been a central issue in inflammatory bowel disease (IBD) for the last years, and has been proposed to be included as the new treatment goal in IBD. The molecular understanding of both the disruption and the healing of the intestinal epithelial cell lining and the mucosal barrier in IBD is complex and only partly understood. There is no general agreement on how to define healed mucosa, but there is a general acceptance that clinicians should use endoscopy and imaging technique in their assessments. Mucosal healing is an old concept that has been actualized in the present era of the highly effective biological agents. Randomized clinical studies with mucosal healing as end-point parameters have been reported, and early mucosal healing has been associated with low complication rates. We are waiting for documentation of whether treatment to healed mucosa can change the natural course of IBD. The concept of immunological remission has recently been introduced and can be the new treatment goal and one of several criteria for discontinuation of biological treatment in IBD. In conclusion, mucosal healing is a fairly novel concept and goal for biological treatment of IBD. There is a need for a standardization of its assessment and validation of the prognostic value.


Crystal structure of Bacillus anthracis virulence regulator AtxA and effects of phosphorylated histidines on multimerization and activity.
The Bacillus anthracis virulence regulator AtxA controls transcription of the anthrax toxin genes and capsule biosynthetic operon. AtxA activity is elevated during growth in media containing glucose and CO(2)/bicarbonate, and there is a positive correlation between the CO(2)/bicarbonate signal, AtxA activity and homomultimerization. AtxA activity is also affected by phosphorylation at specific histidines. We show that AtxA crystallizes as a dimer. Distinct folds associated with predicted DNA-binding domains (HTH1 and HTH2) and phosphoenolpyruvate: carbohydrate phosphotransferase system-regulated domains (PRD1 and PRD2) are apparent. We tested AtxA variants containing single and double phosphomimetic (His→Asp) and phosphoablative (His→Ala) amino acid changes for activity in B. anthracis cultures and for protein-protein interactions in cell lysates. Reduced activity of AtxA H199A, lack of multimerization and activity of AtxAH379D variants, and predicted structural changes associated with phosphorylation support a model for control of AtxA function. We propose that (i) in the AtxA dimer, phosphorylation of H199 in PRD1 affects HTH2 positioning, influencing DNA-binding; and (ii) phosphorylation of H379 in PRD2 disrupts dimer formation. The AtxA structure is the first reported high-resolution full-length structure of a PRD-containing regulator, and can serve as a model for proteins of this family, especially those that link virulence to bacterial metabolism.


A familial syndrome of progressive cone dystrophy, degenerative liver disease, endocrine dysfunction and hearing defect. I. Ophthalmological findings.
Seven patients, 6 females and one male, with progressive cone dystrophy are reported. One patient developed amaurosis in one eye and fere amaurosis in the other. The least affected patient (13 years of age) had fairly good central cone vision, but a rod response only outside the central area. Attenuated retinal vessels, disc pallor and general atrophic appearance without pigmentation were typical findings. Six of the patients originated from 2 sibships. Increasing impairment of vision during pregnancy was seen in two patients. Pathological glucose tolerance, diabetes, liver disease, endocrinological disturbances, and hearing defects were recorded. Thus, this cone dystrophy appears to be part of a disease affecting several organs. The familial occurrence suggests that this disorder is inherited.


Long-term evolution and growth patterns in a family with Bartter's syndrome.
The five-year evolution of three siblings of a sibship affected by a combined potassium and magnesium wasting abnormality associated with hypocalciuria is presented. Fractional distal chloride reabsorption was low. A number of therapeutic measures were attempted with partial or no success. Consequently, patients were kept on oral potassium and magnesium supplementation only. Special attention was paid to nutritional status. Although the three girls maintained a plasma potassium range of 2.6 to 3 meq/l, sexual and mental maturation were not delayed. Growth remained within the lower limits of normality. Considering the poor results obtained from treatment and the side-effects, we conclude that this long-term approach is sound, at least, in less severe cases of Bartter's syndrome.


[Delayed brain abscess as a complication of a CSF shunt].
Infection is a common complication of ventriculoperitoneal shunts, but the formation of a brain abscess related to shunt system is very rare. We present the case of a patient who developed a brain abscess around a ventricular catheter left in place after removing the valvular system. This 38 years old woman, underwent ventriculoperitoneal shunting for hydrocephalus secondary to an acoustic neurinoma. During a period of thirteen years she suffered two episodes of meningitis, and after the second one the CSF shunt was removed excepting for the ventricular catheter which appeared to be adherent to the choroid plexus. One year later, a brain abscess arised around that catheter, and both the abscess and the catheter were removed (Proteus mirabilis was the cultured germ). In spite of the risks related to removal of a proximal catheter adherent to the choroid plexus, the risk linked to the removal of a ventricular catheter must be counterbalanced with the risk of infection.


Differences between attendings' and fellows' perceptions of futile treatment in the intensive care unit at one academic health center: implications for training.
Knowing when patients are too ill to benefit from intensive care is essential for clinicians to recommend aggressive or palliative care as appropriate. To explore prognostic ability among critical care fellows, the authors compared fellows' and attendings' assessments of futile critical care and evaluated factors associated with assessments. Thirty-six attendings and 14 fellows in intensive care units at the University of California, Los Angeles, were surveyed daily for three months (December 2011-March 2012) to identify patients perceived as receiving futile treatment. Frequency of futile treatment assessments and reasons listed by attendings versus fellows were compared. Predictors of futile treatment assessments by provider type were assessed using multivariate probit models. Attendings made 6,897 assessments on 1,125 patients; fellows made 4,407 assessments on 773 patients. Fellows assessed 161 (20.8%) patients as receiving futile treatment, compared with attendings (123 [10.9%] patients, P<.001), and listed fewer reasons that treatment was futile (P<.001). Fellows were more likely to assess a patient as receiving futile treatment by the second day, whereas attendings took four days. Patients assessed as receiving futile treatment by fellows were less likely than patients so assessed by attendings to die in the hospital (51% versus 68%, P=.003) and within six months (62% versus 85%, P<.001). Fellows made earlier assessments and judged more patients to be receiving futile treatment than attendings, and their assessments were less predictive of mortality, suggesting that assessment of treatment appropriateness develops with experience.


Combined radial optic neurotomy, internal limiting membrane peeling, and intravitreal triamcinolone acetonide for central retinal vein occlusion.
To evaluate the efficacy of radial optic neurotomy, internal limiting membrane peeling, and intravitreal triamcinolone acetonide for central retinal vein occlusion. Eight consecutive eyes with central retinal vein occlusion with a duration of less than 6 months, cystoid macular edema, and best-corrected visual acuity (BCVA) of less than 20/200 were enrolled. BCVA, intraocular pressure, fluorescein angiography, and optical coherence tomography were evaluated. After 4.75 +/- 0.7 months, BCVA significantly improved, intraocular pressure was well controlled, and fluorescein angiography showed perfused state and reduction of the number of retinal hemorrhages in all eyes. Optical coherence tomography revealed significant reduction of macular thickness. Bleeding in the neurotomy site occurred in 3 cases. Radial optic neurotomy, internal limiting membrane peeling, and intravitreal triamcinolone acetonide may be useful in the management of central retinal vein occlusion with macular edema.


Synergetic spin crossover and fluorescence in one-dimensional hybrid complexes.
Hybrid materials integrated with a variety of physical properties, such as spin crossover (SCO) and fluorescence, may show synergetic effects that find applications in many fields. Herein we demonstrate a promising post-synthetic approach to achieve such materials by grafting fluorophores (1-pyrenecarboxaldehyde and Rhodamine B) on one-dimensional SCO Fe(II) structures. The resulting hybrid materials display expected one-step SCO behavior and fluorescent properties, in particular showing a coupling between the transition temperature of SCO and the temperature where the fluorescent intensity reverses. Consequently, synergetic effect between SCO and fluorescence is incorporated into materials despite different fluorophores. This study provides an effective strategy for the design and development of novel magnetic and optical materials.


Appearance of substance P-like immunoreactive neurons in the rat superior colliculus after neonatal eye enucleation.
The possibility that eye enucleation within one day after birth has an effect on the postnatal development of substance P (SP)-like-immunoreactive (SP-I) structures in the superior colliculus (SC) was investigated in the rat. Results were compared with those in animals enucleated at postnatal day 15. All the animals were allowed to survive until postnatal day 90, after which changes in SP-I neurons and fibers were identified immunohistochemically. In colchicine-treated rats, the most remarkable changes occurred in SP-I neurons following eye enucleation at birth; large numbers of SP-I neurons appeared in the ventral part of the stratum griseum superficiale (SGS), stratum opticum (SO) and stratum griseum intermediale (SGI) of the deafferentated SC. SP-I neurons did not appear in these layers, when deafferentation of the SC was carried out in rats at postnatal day 15. These findings suggest strongly that eye enucleation at birth affects the production of SP of neurons in the ventral part of the SGS, SO and SGI at the deafferentated SC. The appearance of SP-I neurons in the neonatal eye enucleation may be due, at least partially, to reorganization of another neuronal system in the SC.


Extended survival in advanced-stage multiple myeloma patients treated with gallium nitrate.
Adjunctive anti-bone resorption therapy has become an important part of multiple myeloma (MM) treatment. Currently, no definite evidence exists that this therapy increases survival. We examined a cohort of 13 of 167 patients (8%) treated with the M-2 protocol who received adjuvant gallium nitrate (GN) treatment for osteolysis, and then compared the outcome of these patients to all individuals treated with the M-2 regimen. The stage at diagnosis was IA in two patients, IIIA in 10 patients and IIIB in one. Median age at diagnosis was 51 years (range 35-73). Median (range) of entry data were: paraprotein level: 6000mg/dl (3058-8675); beta2M: 2.7mg/l, (1.2-9.6); LDH: 166U/l (142-237); hemoglobin: 10.2 g/dl (8.3-12.6); albumin: 3.7 g/dl (2.5-5.0); calcium: 10.0 mg/dl (8.3-14.5); creatinine: 1.2 mg/dl (0.7-2.7). Median survival in these patients was 87+ months from time of diagnosis compared with 48 months for all other patients treated on the M-2 protocol. We identified a subgroup of patients with remarkably prolonged survival who had received M-2 chemotherapy and GN. Survival in this group markedly exceeds expectations for patients with advanced stage disease and poor prognostic features. The administration of GN may have a positive impact on survival either by decreasing skeletal complications or through a direct action of GN on the complex cytokine network involved in the proliferation of the malignant myeloma cell.


RNase III cleavage is obligate for maturation but not for function of Escherichia coli pre-23S rRNA.
RNase III makes the initial cleavages that excise Escherichia coli precursor 16S and 23S rRNA from a single large primary transcript. In mutants deficient in RNase III, no species cleaved by RNase III are detected and the processing of 23S rRNA precursors to form mature 23S rRNA fails entirely. Instead, 50S ribosomes are formed with rRNAs up to several hundred nucleotides longer than mature 23S rRNA. Unexpectedly, these aberrant subunits function well enough to participate in protein synthesis and permit cell growth. Consistent with the inference that RNase III cleavages are absolutely required for 23S rRNA maturation, when 50S ribosomes from a strain deficient in RNase III were incubated with a ribosome-free extract from a RNase III+ strain, rRNA species processed by RNase III and species with normal mature 23S rRNA termini were produced.


Biotransformation of waste lignin products by the soil-inhabiting yeast Trichosporon pullulans.
In this study the biotransformation of lignin by-products of beechwood pulping with a soil-inhabiting yeast strain of Trichosporon pullulans was examined. The structural and molecular changes in the lignin during a cultivation process were determined by 13C NMR spectroscopy and gel permeation chromatography analysis, which confirmed the ability of the yeast strain tested to biodegrade lignin. Enzymatic analysis showed the presence of lignin peroxidase and Mn(II) peroxidase in the culture supernatant. The ligninolytic activity of both enzymes increased under carbon-depleted conditions. This observation is particularly important in the biodegradation of recalcitrant lignins in soil.


The full phase space dynamics of a magnetically levitated electromagnetic vibration harvester.
We consider the motion of an electromagnetic vibrational energy harvester (EMVEH) as function of the initial position and velocity and show that this displays a classical chaotic dynamical behavior. The EMVEH considered consists of three coaxial cylindrical permanent magnets and two coaxial coils. The polarities of the three magnets are chosen in such a way that the central magnet floats, with its lateral motion being prevented by enclosion in a hollow plastic tube. The motion of the floating magnet, caused by e.g. environmental vibrations, induces a current in the coils allowing electrical energy to be harvested. We analyze the behavior of the system using a numerical model employing experimentally verified expressions of the force between the magnets and the damping force between the floating magnet and the coils. We map out the phase space of the motion of the system with and without gravity, and show that this displays a fractal-like behavior and that certain driving frequencies and initial conditions allow a large power to be harvested, and that more stable states than two exists. Finally, we show that at leasts fifth order polynomial approximation is necessary to approximate the magnet-magnet force and correctly predict the system behavior.


Characterization of fast pyrolysis bio-oils produced from pretreated pine wood.
The pretreatment of biomass prior to the fast pyrolysis process has been shown to alter the structure and chemical composition of biomass feed stocks leading to a change in the mechanism of biomass thermal decomposition. Pretreatment of feed stocks prior to fast pyrolysis provides an opportunity to produce bio-oils with varied chemical composition and physical properties. This provides the potential to vary bio-oil chemical and physical properties for specific applications. To determine the influence of biomass pretreatments on bio-oil produced during fast pyrolysis, we applied six chemical pretreatments: dilute phosphoric acid, dilute sulfuric acid, sodium hydroxide, calcium hydroxide, ammonium hydroxide, and hydrogen peroxide. Bio-oils were produced from untreated and pretreated 10-year old pine wood feed stocks in an auger reactor at 450 degrees C. The bio-oils' physical properties of pH, water content, acid value, density, viscosity, and heating value were measured. Mean molecular weights and polydispersity were determined by gel permeation chromatography. Chemical characteristics of the bio-oils were determined by gas chromatography-mass spectrometry and Fourier transform infrared techniques. Results showed that the physical and chemical characteristics of the bio-oils produced from pretreated pine wood feed stocks were influenced by the biomass pretreatments applied. These physical and chemical changes are compared and discussed in detail in the paper.


Rv3351c, a Mycobacterium tuberculosis gene that affects bacterial growth and alveolar epithelial cell viability.
Despite the interactions known to occur between various lower respiratory tract pathogens and alveolar epithelial cells (AECs), few reports examine factors influencing the interplay between Mycobacterium tuberculosis bacilli and AECs during infection. Importantly, in vitro studies have demonstrated that the M. tuberculosis hbha and esxA gene products HBHA and ESAT6 directly or indirectly influence AEC survival. In this report, we identify Rv3351c as another M. tuberculosis gene that impacts the fate of both the pathogen and AEC host. Intracellular replication of an Rv3351c mutant in the human AEC type II pneumocyte cell line A549 was markedly reduced relative to the complemented mutant and parent strain. Deletion of Rv3351c diminished the release of lactate dehydrogenase and decreased uptake of trypan blue vital stain by host cells infected with M. tuberculosis bacilli, suggesting attenuated cytotoxic effects. Interestingly, an isogenic hbha mutant displayed reductions in AEC killing similar to those observed for the Rv3351c mutant. This opens the possibility that multiple M. tuberculosis gene products interact with AECs. We also observed that Rv3351c aids intracellular replication and survival of M. tuberculosis in macrophages. This places Rv3351c in the same standing as HBHA and ESAT6, which are important factors in AECs and macrophages. Defining the mechanism(s) by which Rv3351c functions to aid pathogen survival within the host may lead to new drug or vaccine targets.


Interleukin 2 regulates immune interferon (IFN gamma) production by normal and suppressor cell cultures.
A Lyt-2+ lymphocyte is responsible for immune interferon (IFN gamma) production in mitogen-stimulated mouse spleen cell cultures. A Lyt-1+ helper cell is required for the induction of IFN gamma. Interleukin 2 (IL 2) can specifically replace the Lyt 1 cell requirement. IL 2 will also abrogate suppressor cell inhibition of IFN gamma production. IFN gamma production appears to be regulated by a dynamic interaction between helper cells (source of IL 2), suppressor cells (absorption of IL 2?) and IFN gammna-producer cells.


Optimized regulation of gene expression using artificial transcription factors.
A major focus in the basic science of gene therapy is the study of factors involved in target-specific regulation of gene expression. Optimization of artificial or "designer" transcription factors capable of specific regulation of target genes is a prerequisite to developing practical applications in human subjects. In this paper, we present a systematic and combinatorial approach to optimize engineered transcription factors using designed zinc-finger proteins fused to transcriptional effector domains derived from the naturally occurring activators (VP16 or P65) or repressor (KRAB) proteins. We also demonstrate effective targeting of artificial transcription factors to regulate gene expression from three different constitutive viral promoters (SV40, CMV, RSV). Achieving a desired level of gene expression from a targeted region depended on several variables, including target site affinities for various DNA-binding domains, the nature of the activator domain, the particular cell type used, and the position of the target site with respect to the core promoter. Hence, several aspects of the artificial transcription factors should be simultaneously evaluated to ensure the optimum level of gene expression from a given target site in a given cell type. Our observations and our optimization approach have substantial implications for designing safe and effective artificial transcription factors for cell-based and therapeutic uses.


New evidence for the structure of myxinol.
1. Preliminary spectroscopic examination of a second component of hagfish bile salts suggested that it might be 3beta,7alpha,26(27)-trihydroxy-5alpha-cholestane. 2. Impure reduction products of the 3beta,26(27)-dihydroxycholestane-7,16-dione previously made from myxinol disulphate appeared also to have the 5alpha-configuration. 3. Infrared, nuclear-magnetic-resonance and mass-spectrographic as well as optical-rotatory-dispersion measurements on 3beta,26(27)-dihydroxycholestane-7,16-dione showed that it was a 5alpha-compound. 4. Myxinol is thus 3beta,7alpha,16alpha,26(27)-tetrahydroxy-5alpha-cholestane; new nuclear-magnetic-resonance measurements on myxinol tetra-acetate at higher resolution confirm this structure.


Stability of fluorouracil in plastic containers used for continuous infusion at home.
The stability of fluorouracil when prepared and stored in plastic containers used for continuous infusion in home therapy was studied. Fluorouracil was diluted with 5% dextrose injection. Samples were stored for 16 weeks at 5 degrees C and room temperature in polyvinyl chloride drug reservoirs and at 5 degrees C in elastomeric balloons used in a disposable drug pump. Periodically, samples were tested by a stability-indicating high-performance liquid chromatography (HPLC) assay. At 5 degrees C, there was no significant degradation. At room temperature, there was a progressive increase in fluorouracil concentration, presumably because of evaporation of water. A 10% change in the fluorouracil concentration over 55 days was predicted. Fluorouracil is stable in 5% dextrose injection for at least 16 weeks when stored at 5 degrees C in the two types of containers studied.


Body mass index, weight loss and energy intake of old Danish nursing home residents and home-care clients.
Low body mass index (BMI) and weight loss is common among old home-care clients and nursing home residents in many countries - often in spite of an apparent sufficient intake of energy. The aim of this study was to assess whether a similar problem exists in Denmark. Information regarding height, body weight, BMI, and energy intake was collected. Besides this, 24 residents were weighed regularly during 1 year. The study was carried out in five nursing homes and two home-care districts situated in the area of Copenhagen. A total of 180 residents and 200 clients 65+ years of age participated. Main outcome measures were prevalence of BMI < 20 kg/m2, BMI < 18.5 kg/m2, acute weight loss and median energy intake. Thirty-three and thirty per cent of the residents and clients had a BMI < 20 kg/m2 and, 22 and 12% had a BMI < 18.5 kg/m2. Of the residents, who were weighed regularly, 38% had a weight loss above 5%. The median intake of energy was equal to or above the estimated median energy requirement. Hence, the prevalence of low BMI and weight loss among old Danish nursing home residents and home-care clients is high and similar to that found in other countries. There is a need for increased focus on nutritional state and risk factors in these settings.


[Biophysical properties of membrane dressing made of bacterial cellulose].
In the paper, review of papers devoted to biophysical properties of membrane dressing made of bacterial cellulose was done. These properties were determined on the basis of studies on osmotic and diffusive transport through pure (non modified) bacterial cellulose membrane form called Bio-Fill. The measures of these properties are values of membrane transport parameters resulted from Kedem-Katchalsky's theory and interferograms of near-membrane regions made laser interferometric method.


Simultaneous measurement of nineteen binding constants of peptides to vancomycin using affinity capillary electrophoresis-mass spectrometry.
On-line affinity capillary electrophoresis-electrospray ionization-mass spectrometry (ACE-MS) was used for the simultaneous measurement of multiple binding constants of an all-D-tetrapeptide library to the model receptor, vancomycin. Determination of Kd values for the 19 peptides of the form Fmoc-DXYA is demonstrated. The data are compared with the results obtained for individual compounds using ACE-UV, and good correlation between the two detection methods is shown. Simultaneous determination of multiple Kd values by ACE-MS is achieved in one set of experiments, whereas only one Kd value can be obtained by ACE-UV during the same time. ACE-MS measures multiple binding constants in solution in a fast and reliable manner using femtomole amounts of samples.


Enhanced malignant phenotype of urethane-induced lung adenoma associated with sialoadenectomy in BALB/c mice.
Mouse lung adenoma were induced in adult female BALB/c mice by chronic i.p. urethane injection. Sialoadenectomy of the mice prior to carcinogen treatment did not alter the frequency or size of lung adenoma. However, sialoadenectomized mice exhibited a decrease in the proportion (18 versus 52%) of tumours which exhibited a low nuclear to cytoplasmic ratio and which grew along alveolar septa. Sialoadenectomy was also related to a complementary increase in the proportion (82 versus 48%) of tumours with large vesicular nuclei, prominent nucleoli and generally increased phenotypic features of malignancy. Replacement of epidermal growth factor (EGF) in sialoadenectomized mice restored the tumour-type ratio observed in non-sialoadenectomized mice. These data are discussed with respect to the possible roles of EGF in mouse lung alveologenic carcinoma formation and the cell type of origin for the tumours observed.