Assessment of reward responsiveness in the response bias probabilistic reward task in rats: implications for cross-species translational research.
Mood disorders, such as major depressive disorder, are characterized by abnormal reward responsiveness. The Response Bias Probabilistic Reward Task (hereafter referred to as probabilistic reward task (PRT)) quantifies reward responsiveness in human subjects, and an equivalent animal assessment is needed to facilitate preclinical translational research. Thus, the goals of the present studies were to develop, validate and characterize a rat analog of the PRT. Adult male Wistar and Long-Evans rats were trained in operant testing chambers to discriminate between two tone stimuli that varied in duration (0.5 and 2 s). During a subsequent test session consisting of 100 trials, the two tones were made ambiguous (0.9 and 1.6 s) and correct identification of one tone was reinforced with a food pellet three times more frequently than the other tone. In subsequent experiments, Wistar rats were administered either a low dose of the dopamine D2/D3 receptor agonist pramipexole (0.1 mg kg(-1), subcutaneous) or the psychostimulant amphetamine (0.5 mg kg(-1), intraperitoneal) before the test session. Similar to human subjects, both rat strains developed a response bias toward the more frequently reinforced stimulus, reflecting robust reward responsiveness. Mirroring prior findings in humans, a low dose of pramipexole blunted response bias. Moreover, in rats, amphetamine potentiated response bias. These results indicate that in rats, reward responsiveness can be quantified and bidirectionally modulated by pharmacological manipulations that alter striatal dopamine transmission. Thus, this new procedure in rats, which is conceptually and procedurally analogous to the one used in humans, provides a reverse translational platform to investigate abnormal reward responsiveness across species.


Lyme disease with concurrent ehrlichiosis.
Lyme disease constitutes a major health hazard with an increased incidence throughout the United States, in particular the eastern states. Human ehrlichiosis, also a tick-borne illness, has recently been identified. It is characterized by fever, headache, malaise, leukopenia, thrombocytopenia, and elevated liver enzyme titers, and has been reported to occur mainly in the South Central and South Atlantic states. As with Lyme disease, most patients have a history of tick exposure. These two diseases may be difficult to differentiate clinically. Physicians must consider the possibility of both infections when patients become ill with a systemic illness after tick exposure. Although certain demographic and clinical features are characteristic of these diseases, they can be misleading. Only serologic evidence can confirm the diagnosis. Two cases of concurrent Borrelia and Ehrlichia infections have been previously reported. The authors herein describe a third case that further illustrates the potential diagnostic dilemma posed by the concurrence of these two entities.


Effect of flow rate and insulin priming on the recovery of insulin from microbore infusion tubing.
A retrospective medical record review of 13 consecutive, hyperglycemic, extremely low birth weight (ELBW) infants treated with continuous insulin infusions revealed a 14- to 24-hour delay (mean, 19 hours) in blood glucose normalization despite stepwise increases in insulin infusion rates. This in vitro study examined the effects of flow rate and insulin priming on insulin recovery from polyvinyl chloride (PVC) tubing and polyethylene (PE)-lined PVC tubing infused with a standard insulin stock solution. Stock insulin solution (0.2 U/mL) was infused through microbore PVC or PE-lined tubing at flow rates of 0.05 and 0.2 mL/h. To determine if saturation of nonspecific binding sites would alter effluent insulin concentration, we compared insulin recovery from tubing previously flushed with the stock solution and tubing primed with 5 U/mL of insulin for 20 minutes. Effluent samples, which were collected at baseline and at six time points during a 24-hour period, were immediately frozen at -20 degreesC. Insulin concentration was measured by IMx immunoassay. Data were analyzed using general linear modeling with repeated measures. At 0.05 mL/h flow rate, insulin recovery from unprimed PVC tubing at 1, 2, 4, and 8 hours was 17%, 11%, 27%, and 55%, respectively, with 100% recovery at 24 hours. From insulin-primed tubing, insulin recovery was approximately 70% at 1, 2, and 4 hours, and close to 100% at 8 hours. At a faster flow rate of 0.2 mL/h, insulin recovery at 1, 2, 4, and 8 hours was 22%, 38%, 67%, and 75% vs 42%, 85%, 91% and 95% from unprimed and insulin-primed PVC tubing, respectively. Similar results were obtained from unprimed and insulin-primed PE-lined tubing at 0.2 mL/h flow rate. Priming of microbore tubing with 5 U/mL of insulin solution for 20 minutes to block nonspecific binding sites enhances delivery of a standard insulin stock at infusion rates typically used to treat hyperglycemic ELBW infants. We conclude that priming the tubing with a higher concentration of insulin before initiation of standard insulin infusion therapy should accelerate achievement of steady-state insulin delivery and correction of hyperglycemia in ELBW infants.


Extension of the Advanced REACH Tool (ART) to Include Welding Fume Exposure.
The Advanced REACH Tool (ART) is a mechanistic higher tier model to estimate inhalation exposure to chemicals using a Bayesian approach. Currently the ART model does not include exposure to welding fumes within its applicability domain; it has only been calibrated for vapours, mists, and dusts. To extend the scope to metal fumes it is necessary to review the model structure to ensure that it is appropriate, and to calibrate the updated model using available welding fume exposure measurements. This paper provides a discussion of the key modifying factors (MFs) that should be considered to extend the ART model to include welding fume exposure. Based on our literature review, welding process type, input power level, shield gas, and welding electrodes have important impact on fume formation rates (FFRs). In addition, the convective dispersion of the fume away from the weld and the interaction of the welder with the fume plume should be incorporated into the ART model. Other aspects of the ART, such as the local ventilation, do not require modification to accommodate welding fume exposure. The ART does not include the impact of wearing personal protective equipment and so this is not included in our evaluation. Proposals are made for extending the scope of the ART to include welding processes.


The Pathophysiological Role of Neutrophil Extracellular Traps in Inflammatory Diseases.
Neutrophil pathogen-killing mechanism termed neutrophil extracellular traps (NETs) has been recently identified. NETs consist of chromatin and histones along with serine proteases and myeloperoxidase and are induced by a great variety of infectious and non-infectious stimuli. NETosis is a kind of programmed neutrophil death characterized by chromatin decondensation and release of nuclear granular contents, mainly driven by peptidylarginine deiminase 4 citrullination of histones. Although classically related to the protection against infectious pathogens, nowadays NETs have been described as a player of several pathophysiological processes. Neutrophil dysregulation has been demonstrated in the pathogenesis of most representative vascular diseases, such as acute coronary syndrome, stroke and venous thrombosis. Indeed, NETs have been identified within atherosclerotic lesions and arterial thrombi in both human beings and animal models. Moreover, an imbalance in this mechanism has been proposed as a critical source of modified and/or externalized autoantigens in autoimmune and inflammatory diseases. Finally, an update on the role of NETs in the pathogenesis of cancer has been included. In the present review, based on papers released on PubMed and MEDLINE up to July 2017, we point to update the knowledge on NETs, from their structure to their roles in infectious diseases as well as in cardiovascular diseases, autoimmunity, metabolic disorders and cancer, with a look to future perspectives and therapeutic opportunities.


Determinants of the Efficacy of Natural Selection on Coding and Noncoding Variability in Two Passerine Species.
Population genetic theory predicts that selection should be more effective when the effective population size (Ne) is larger, and that the efficacy of selection should correlate positively with recombination rate. Here, we analyzed the genomes of ten great tits and ten zebra finches. Nucleotide diversity at 4-fold degenerate sites indicates that zebra finches have a 2.83-fold larger Ne. We obtained clear evidence that purifying selection is more effective in zebra finches. The proportion of substitutions at 0-fold degenerate sites fixed by positive selection (α) is high in both species (great tit 48%; zebra finch 64%) and is significantly higher in zebra finches. When α was estimated on GC-conservative changes (i.e., between A and T and between G and C), the estimates reduced in both species (great tit 22%; zebra finch 53%). A theoretical model presented herein suggests that failing to control for the effects of GC-biased gene conversion (gBGC) is potentially a contributor to the overestimation of α, and that this effect cannot be alleviated by first fitting a demographic model to neutral variants. We present the first estimates in birds for α in the untranslated regions, and found evidence for substantial adaptive changes. Finally, although purifying selection is stronger in high-recombination regions, we obtained mixed evidence for α increasing with recombination rate, especially after accounting for gBGC. These results highlight that it is important to consider the potential confounding effects of gBGC when quantifying selection and that our understanding of what determines the efficacy of selection is incomplete.


Symptomatic and neurophysiological responses of paclitaxel- or cisplatin-induced neuropathy to oral acetyl-L-carnitine.
Acetyl-L-carnitine (ALC) improves non-oncological neuropathies. We tested oral ALC (1 g tid) for 8 weeks in 25 patients with neuropathy grade 3 (common toxicity criteria--CTC) during paclitaxel or cisplatin therapy, or grade 2 persisting for at least three months after discontinuing the drugs. An independent neurologist assessed patients before and after ALC. All patients except one reported symptomatic relief, and only two described grade 1 nausea. The sensory neuropathy grade improved in 15 of 25 (60%), and motor neuropathy in 11 of 14 patients (79%). Total neuropathy score (TNS) that included neurophysiological measures improved in 23 (92%). Amelioration of sensory amplitude and conduction velocity (sural and peroneal nerves) was measured in 22 and 21 patients, respectively. Symptomatic improvement persisted in 12 of 13 evaluable patients at median 13 months after ALC. In view of its effect in improving established paclitaxel- and cisplatin-neuropathy, we recommend ALC testing in preventing progression or revert symptoms during neurotoxic chemotherapy.


Abuse and clinical value of diuretics in eating disorders therapeutic applications.
Diuretic abuse as a means of purging is common in patients with bulimia nervosa. We sought to illustrate the pathophysiologic effects of diuretics and purging on a patient with bulimia nervosa's fluid and electrolyte status and to clarify the role of diuretics in the management of volume status during refeeding. We reviewed the literature pertaining to diuretic abuse, purging, bulimia nervosa, and diuretic therapy. Purging behaviors lead to volume depletion and a state of heightened aldosterone production. Patients with bulimia nervosa commonly undergo rapid rehydration with intravenous fluid administration. In the setting of hyperaldostreronism, aggressive rehydration leads to avid salt retention and the development of marked amounts of edema. Providers should understand both the background renal pathophysiology of the patient with bulimia nervosa and the mechanisms of action of diuretics to correctly use diuretics as focused therapeutic agents for this patient population.


ladybird, a tandem of homeobox genes that maintain late wingless expression in terminal and dorsal epidermis of the Drosophila embryo.
ladybird early and ladybird late genes, tandemly located in the Drosophila 93E homeobox gene cluster, encode highly related homeodomain-containing transcription factors. Here we report the cloning of the complete cDNA sequences of both genes and a study of their expression and regulatory interactions with the segment polarity gene wingless in the epidermis. ladybird genes are co-expressed with wingless in epidermal cells close to the posterior parasegmental boundaries and in terminal regions of the body. In mutant embryos with altered wingless function, transcription of ladybird early and ladybird late is changed; it disappears completely from the epidermis in wingless-embryos, indicating wingless-dependence. After 6 hours of development, wingless expression is maintained by gooseberry in the ventral epidermis. However, in the dorsal epidermis and the terminal regions of the body, expression of wingless is independent of gooseberry but requires a wingless-ladybird regulatory feedback loop. Loss of ladybird function reduces the number of wingless-expressing cells in dorsal epidermis and leads to complete inactivation of wingless in the anal plate. Consequently, mutant ladybird embryos fail to develop anal plates and ubiquitous embryonic expression of either one or both ladybird genes leads to severe defects of the dorsal cuticle. Lack of late wingless expression and anal plate formation can be rescued with the use of a heat-shock-ladybird transgene.


Herbal medicine use during pregnancy in a group of Australian women.
There are limited data on the extent of women's use of herbal medicines during pregnancy, despite the fact that knowledge of the potential benefits or harms of many of these products is sparse, particularly with respect to their use in pregnancy. We aimed to measure the prevalence of herbal medicine use in a group of pregnant women attending a public tertiary maternity hospital in Melbourne, Australia. Secondary aims were to explore why women took the herbal medicine, where they received advice, what form the supplements took and if they perceived the supplements to be helpful. Consecutive pregnant women were approached in the antenatal clinic and the birth centre at around 36-38 weeks gestation. A questionnaire was developed and self-administered in English, as well as being translated into the four most common languages of women attending the hospital: Cantonese, Vietnamese, Turkish and Arabic. Back translation into English was undertaken by different professional translators to verify accuracy of both words and concepts. Data collected included demographic information, model of pregnancy care and herbal supplement use. Descriptive statistics were used initially, with stratified and regression analysis to compare sub-groups. Of 705 eligible women, 588 (83%) agreed to participate. Of these, 88 (15%) completed the questionnaire in a language other than English. Thirty-six percent of women took at least one herbal supplement during the current pregnancy. The most common supplements taken were raspberry leaf (14%), ginger (12%) and chamomile (11%). Women were more likely to take herbal supplements if they were older, tertiary educated, English speaking, non-smokers and primiparous. Use of herbal supplements in pregnancy is likely to be relatively high and it is important to ascertain what supplements (if any) women are taking. Pregnancy care providers should be aware of the common herbal supplements used by women, and of the evidence regarding potential benefits or harm.


Survey of surgical emergencies in a rural population in the Northern Areas of Pakistan.
To determine the incidence of specific surgical emergencies in a mountainous rural community in the Northern Areas of Pakistan and to assess use of existing health services, and outcomes related to acute surgical illness events. A cross-sectional population-based survey was conducted. Interviewers visited each of 118 villages in the study area (population 100000), selected a random sample from a total of 9900 households, and interviewed the oldest premenopausal female member (n = 836). Questions were focused on injury, acute abdomen, and/or maternal morbidity occurring in the past year. Cases were included as surgical emergencies when one or more index clinical features indicating a potential for surgical intervention were present. Mortality from a wider range of surgical emergencies was also elicited based on the respondent's lifetime knowledge of the household. The incidence rates were 1531/100000 persons per year for injuries; 1364/100000 for acute abdomen, and 16462/100000 for maternal morbidity. The rate of injuries was 2.7 times higher and that of acute abdomen twice as high in males as in females. The injury rate decreased with advancing age, being 13 times higher in children < 5 years than in adults > 40. By contrast, the rate for acute abdomen showed a rise with advancing age, being 8 times higher in the > 40 age group than in under-fives. Burns, falls and road accidents, in that order, were the commonest forms of injury accounting for 82% of 138 cases. Of 43 burn casualties, 46% were in the age group < 5 years; there was no gender bias. Of 71 casualties from falls and road accidents, 85% were aged 6-40 years; there was 6 : 1 male predominance. The maternal morbidity rate was highest in the age group 25-35 years and may be attributed to the high pregnancy rate in this age bracket. Of 408 patients with acute surgical illness, 85% were managed initially at home or close to home in a health centre, dispensary or civil hospital; 32% eventually sought specialist surgical care. The overall rate for minor and major surgical procedures was 411/100000 persons per year (lowest estimate), and appeared to be low. The rate of operative deliveries at 11.8/1000 deliveries (lowest estimate) was particularly low. The mortality rates were correspondingly high: 55/100000 persons per year for injuries and for acute abdomen (lowest estimates). The maternal mortality ratio was particularly high at 8.9/1000 deliveries (lowest estimate). Annual mortality rates derived from deaths recalled during the respondent's lifetime in the household (mean period = 26 years), tended to corroborate the results of the 1-year survey. The incidence rates for broad categories of serious acute surgical illness in the study population far exceeded the rates of acute surgical intervention. Mortality rates were correspondingly high. Such evidence points to a large unmet surgical need and ought to spur improvements in the health service.


Reduced Position Effect in Mature Transgenic Plants Conferred by the Chicken Lysozyme Matrix-Associated Region.
Matrix-associated regions may be useful for studying the role of chromatin architecture in transgene activity of transformed plants. The chicken lysozyme A element was shown to have specific affinity for tobacco nuclear matrices, and its influence on the variability of transgene expression in tobacco plants was studied. T-DNA constructs in which this element flanked either the [beta]-glucuronidase (GUS) reporter gene or both reporter and selection gene were introduced in tobacco. The variation in GUS gene activity was reduced significantly among mature first-generation transgenic plants carrying the A element. Average GUS activity became somewhat higher, but the maximum attainable level of gene expression was similar for all three constructs. Transient gene expression assays showed that the A element did not contain general enhancer functions; therefore, its presence seemed to prevent the lower levels of transgene expression. The strongest reduction in variability was found in plants transformed with the construct carrying the A elements at the borders of the T-DNA. In this population, expression levels became copy number dependent. The presence of two A elements in the T-DNA did not interfere with meiosis.


Spatial-temporal variability in diazotroph assemblages in Chesapeake Bay using an oligonucleotide nifH microarray.
The distribution of nitrogen-fixing microorganisms in the Chesapeake Bay was investigated using fingerprints from a nifH microarray comprised of 706 60-mer oligonucleotide nifH probes representing cultivated organisms and environmental clones from different nifH clusters. Diverse nifH targets, amplified from samples using degenerate nifH primers, were detected in water column and sediment samples collected in April and October, 2001-2002. Total nifH richness and diversity (Simpson's and Shannon indices) were highest at the most riverine, oligohaline North Bay station. In most samples, the highest diversity was in nifH Cluster 3, which includes many anaerobes, while Cluster 1 (alpha-, beta- gamma- Proteobacteria, Cyanobacteria) targets had the greatest microarray signal intensities. In a multidimensional scaling analysis, deep water communities from April and October were similar within each of the sampling sites, while the surface communities had more variability. Diazotroph communities in the water column in the North Bay were distinct from the Mid- and South Bay communities, and there was a gradual change in sediment diazotroph assemblages from the North to the South Bay. Diazotrophic assemblages from the majority of the water column samples from the Mid- and South Bay clustered with the sediment assemblage in Mid-Bay. Dissolved inorganic nitrogen, salinity, dissolved organic carbon and dissolved organic phosphorus had a significant relationship with the diazotrophic bacterioplankton community. Higher diversity in the freshwater end of the system may reflect variability in disturbance rates and environmental conditions such as forms and concentrations of organic matter, nutrients and oxygen.


Local structural differences in homologous proteins: specificities in different SCOP classes.
The constant increase in the number of solved protein structures is of great help in understanding the basic principles behind protein folding and evolution. 3-D structural knowledge is valuable in designing and developing methods for comparison, modelling and prediction of protein structures. These approaches for structure analysis can be directly implicated in studying protein function and for drug design. The backbone of a protein structure favours certain local conformations which include α-helices, β-strands and turns. Libraries of limited number of local conformations (Structural Alphabets) were developed in the past to obtain a useful categorization of backbone conformation. Protein Block (PB) is one such Structural Alphabet that gave a reasonable structure approximation of 0.42 Å. In this study, we use PB description of local structures to analyse conformations that are preferred sites for structural variations and insertions, among group of related folds. This knowledge can be utilized in improving tools for structure comparison that work by analysing local structure similarities. Conformational differences between homologous proteins are known to occur often in the regions comprising turns and loops. Interestingly, these differences are found to have specific preferences depending upon the structural classes of proteins. Such class-specific preferences are mainly seen in the all-β class with changes involving short helical conformations and hairpin turns. A test carried out on a benchmark dataset also indicates that the use of knowledge on the class specific variations can improve the performance of a PB based structure comparison approach. The preference for the indel sites also seem to be confined to a few backbone conformations involving β-turns and helix C-caps. These are mainly associated with short loops joining the regular secondary structures that mediate a reversal in the chain direction. Rare β-turns of type I' and II' are also identified as preferred sites for insertions.


Lasers for vascular lesions: standard guidelines of care.
Lasers are a good therapeutic tool for congenital and acquired vascular lesions. Technological advances in lasers have reduced the adverse effects and increased the efficacy. MACHINES: Among the various lasers used for treating vascular lesions, pulsed dye laser (PDL) has the best efficacy and safety data. The other machines that are widely available are Nd:YAG laser and intense pulse light (IPL). RATIONALE AND SCOPE OF GUIDELINE: Much variation exists in different machines and techniques, and therefore, establishing standard guidelines has limitations. The guidelines recommended here indicate minimum standards of care for lasers on vascular lesions based on current evidence. Laser may be administered by a dermatologist, who has received adequate background training in lasers during post-graduation or later at a center that provides education and training in lasers, or in focused workshops, which provide such trainings. He/she should have adequate knowledge of the lesions being treated, machines, parameters, cooling systems, and aftercare. The procedure may be performed in the physician's minor procedure room with adequate laser safety measures. PWS, hemangioma, facial telangiectasia, rosacea, spider angioma, pyogenic granuloma, venous lakes, leg veins. Absolute: Active local infection, photo-aggravated skin diseases, and medical conditions. Relative: Unstable vitiligo, psoriasis, keloid and keloidal tendencies, patient on isotretinoin, patient who is not cooperative or has unrealistic expectation. Patient selection should be done after detailed counseling with respect to the course of lesions, different treatment options, possible results, cost, need for multiple treatments, and possible postoperative complications. TREATMENT SESSIONS: The number of treatments per lesion varies from 2 to 12 or more at 6-8 week intervals. All lesions may not clear completely even after multiple sessions in many cases. Hence, a realistic expectation and proper counseling is very important. Laser parameters vary with area, type of lesion, skin color, depth of the lesion, and machine used. A test spot may be performed to determine individual specifications. Pain, edema, purpura, bleeding, scarring, postinflammatory hyperpigmentation/hypopigmentation, and atrophy changes.


Detecting circulating antibodies by controlled surface modification with specific target proteins: Application to malaria.
Sensitive detection of specific antibodies by biosensors has become of major importance for monitoring and controlling epidemics. Here we report a development of a biosensor able to specifically measure antibodies in a drop of unmodified blood serum. Within minutes, the detection system measures presence of antibodies against Plasmodium vivax, a causing agent for malaria. The biosensor consists of a layer of carbon nanotubes (CNTs) which were casted on a carbon working electrode area of a three-electrode system and oxidized. An amine layer was produced next by modifying the surface with EDAC/NHS followed by reaction with a diamine compound. Finally, the protein fragments derived from P. vivax containing well-known antigen sequences were casted on this layer and bound through electrostatic interactions, involving hydrogen and ionic bonding. All these chemical changes occurring at the carbon surface along the biosensor assembly were followed and confirmed by Fourier Transformed Infrared s pectrometry (FTIR) and Raman spectroscopy. The presence of antibodies in serum was detected by monitoring the electrical properties of the layer, making use of cyclic voltammetry (CV), electrochemical impedance spectroscopy (EIS) and square wave voltammetry (SWV), against a standard iron probe. Overall, the charge-transfer resistance decreased after antibody binding, because there was an additional amount of protein bound to the surface. This hindered the access of the iron redox probe to the conductive support at the electrode surface. Electrical changes could be measured at antibody concentration as low as ~6-50pg/L (concentrations in the range of 10-15M) and as high as ~70μg/L. Specific measurement with low background was even possible in undiluted serum. Hence, this novel biosensor allows assessing serum antibody levels in real time and in un-manipulated serum samples on-site where needed.


Genome-wide identification of transcriptional start sites in the haloarchaeon Haloferax volcanii based on differential RNA-Seq (dRNA-Seq).
Differential RNA-Seq (dRNA-Seq) is a recently developed method of performing primary transcriptome analyses that allows for the genome-wide mapping of transcriptional start sites (TSSs) and the identification of novel transcripts. Although the transcriptomes of diverse bacterial species have been characterized by dRNA-Seq, the transcriptome analysis of archaeal species is still rather limited. Therefore, we used dRNA-Seq to characterize the primary transcriptome of the model archaeon Haloferax volcanii. Three independent cultures of Hfx. volcanii grown under optimal conditions to the mid-exponential growth phase were used to determine the primary transcriptome and map the 5'-ends of the transcripts. In total, 4749 potential TSSs were detected. A position weight matrix (PWM) was derived for the promoter predictions, and the results showed that 64 % of the TSSs were preceded by stringent or relaxed basal promoters. Of the identified TSSs, 1851 belonged to protein-coding genes. Thus, fewer than half (46 %) of the 4040 protein-coding genes were expressed under optimal growth conditions. Seventy-two percent of all protein-coding transcripts were leaderless, which emphasized that this pathway is the major pathway for translation initiation in haloarchaea. A total of 2898 of the TSSs belonged to potential non-coding RNAs, which accounted for an unexpectedly high fraction (61 %) of all transcripts. Most of the non-coding TSSs had not been previously described (2792) and represented novel sequences (59 % of all TSSs). A large fraction of the potential novel non-coding transcripts were cis-antisense RNAs (1244 aTSSs). A strong negative correlation between the levels of antisense transcripts and cognate sense mRNAs was found, which suggested that the negative regulation of gene expression via antisense RNAs may play an important role in haloarchaea. The other types of novel non-coding transcripts corresponded to internal transcripts overlapping with mRNAs (1153 iTSSs) and intergenic small RNA (sRNA) candidates (395 TSSs). This study provides a comprehensive map of the primary transcriptome of Hfx. volcanii grown under optimal conditions. Fewer than half of all protein-coding genes have been transcribed under these conditions. Unexpectedly, more than half of the detected TSSs belonged to several classes of non-coding RNAs. Thus, RNA-based regulation appears to play a more important role in haloarchaea than previously anticipated.


Effects of temporal hydrostatic pressure on tissue-engineered bovine articular cartilage constructs.
The objective of this study was to determine the effects of temporal hydrostatic pressure (HP) on the properties of scaffoldless bovine articular cartilage constructs. The study was organized in three phases: First, a suitable control for HP application was identified. Second, 10 MPa static HP was applied at three different timepoints (6-10 days, 10-14 days, and 14-18 days) to identify a window in construct development when HP application would be most beneficial. Third, the temporal effects of 10-14-day static HP application, as determined in phase II, were assessed at 2, 4, and 8 weeks. Compressive and tensile mechanical properties, GAG and collagen content, histology for GAG and collagen, and immunohistochemistry for collagen types I and II were assessed. When a culture control identified in phase I was used in phase II, HP application from 10 to 14 days resulted in a significant 1.4-fold increase in aggregate modulus, accompanied by an increase in GAG content, while HP application at all timepoints enhanced tensile properties and collagen content. In phase III, HP had an immediate effect on GAG content, collagen content, and compressive stiffness, while there was a delayed increase in tensile stiffness. The enhanced tensile stiffness was still present at 8 weeks. For the first time, this study examined the immediate and long-term effects of HP on biomechanical properties, and demonstrated that HP has an optimal application time in construct development. These findings are exciting as HP stimulation allowed for the formation of robust tissue-engineered cartilage; for example, 10 MPa static HP resulted in an aggregate modulus of 273 +/- 123 kPa, a Young's modulus of 1.6 +/- 0.4 MPa, a GAG/wet weight of 6.1 +/- 1.4%, and a collagen/wet weight of 10.6 +/- 2.4% at 4 weeks.


Reducing the impact of carbapenem-resistant Enterobacteriaceae on vulnerable patient groups: what can be done?
Carbapenem-resistant Enterobacteriaceae (CRE) is a worldwide challenge and associated with a high mortality rate in critically ill patients. This review focused on rapid diagnosis, optimization of antimicrobial therapy, and implication of effective infection control precautions to reduce impact of CRE on vulnerable patients. Several new diagnostic assays have recently been described for the early diagnosis of CRE. Retrospective studies are supportive for colistin plus meropenem combination for the treatment of CRE infections; however, solid evidence is still lacking. Ceftazidime-avibactam may be an effective therapeutic agent for infections caused by carbapenem-hydrolyzing oxacillinase-48 and Klebsiella pneumoniae carbapenamase-producing Enterobacteriaceae, but not for New Delhi metallo-β-lactamase producers. Gastrointestinal screening may permit early identification of patients with CRE infections. There is not enough evidence to recommend selective digestive decontamination for CRE carriers. The information for rapid diagnosis of CRE is accumulating. There are new agents with high in-vitro activity against CRE, but clinical experience is limited to case reports. Active surveillance with a high rate of compliance to basic infection control precautions seems to be the best approach to reduce the impact of CRE on vulnerable patients.


Induction of immunoglobulin secretion in follicular non-Hodgkin's lymphomas: role of immunoregulatory T cells.
B cell neoplasms are clonal expansions of B lymphocytes thought to be frozen at various points along the normal B cell differentiation pathway. We studied cell suspensions from lymph nodes involved by follicular (nodular) non-Hodgkin's lymphoma to determine the capacity of the malignant B cells to secrete immunoglobulin (Ig). Neoplastic B cells from all 14 follicular lymphomas secreted monoclonal immunoglobulin in culture when appropriate signals were provided. In most cases, maximal Ig secretion occurred when autologous T cells were removed by E rosette depletion, replaced with allogeneic normal T cells, and the cultures were exposed to 12-O-tetradecanoylphorbol-13-acetate. Autologous T cells exerted a suppressor effect on Ig secretion in 8/8 cases studied, diminishing the response of the malignant B cells to allogeneic T cells. This suppressor effect did not correlate with the percentage of cells staining with anti-Leu-2a or with "helper-suppressor" (Leu-3a-Leu-2a) ratios of the lymph node T cells. Our findings demonstrate that the arrested differentiation of most follicular lymphomas is reversible and implicate a T cell-mediated host immunoregulatory mechanism affecting Ig secretion in vivo. An additional contribution of our results is the demonstration of a cell culture system for synthesis of sufficient monoclonal Ig for use as an immunogen in production of anti-idiotype antibodies.


Dysfunctional problem-based learning curricula: resolving the problem.
Problem-based learning (PBL) has become the most significant innovation in medical education of the past 40 years. In contrast to exam-centered, lecture-based conventional curricula, PBL is a comprehensive curricular strategy that fosters student-centred learning and the skills desired in physicians. The rapid spread of PBL has produced many variants. One of the most common is 'hybrid PBL' where conventional teaching methods are implemented alongside PBL. This paper contends that the mixing of these two opposing educational philosophies can undermine PBL and nullify its positive benefits. Schools using hybrid PBL and lacking medical education expertise may end up with a dysfunctional curriculum worse off than the traditional approach. For hybrid PBL schools with a dysfunctional curriculum, standard PBL is a cost-feasible option that confers the benefits of the PBL approach. This paper describes the signs of a dysfunctional PBL curriculum to aid hybrid PBL schools in recognising curricular breakdown. Next it discusses alternative curricular strategies and costs associated with PBL. It then details the four critical factors for successful conversion to standard PBL: dealing with staff resistance, understanding the role of lectures, adequate time for preparation and support from the administrative leadership. Hybrid PBL curricula without oversight by staff with medical education expertise can degenerate into dysfunctional curricula inferior even to the traditional approach from which PBL emerged. Such schools should inspect their curriculum periodically for signs of dysfunction to enable timely corrective action. A decision to convert fully to standard PBL is cost feasible but will require time, expertise and commitment which is only sustainable with supportive leadership.


Reduced nitrate level in individuals with hypertension and diabetes.
Nitric oxide (NO) turnover is vital for proper endothelial function to maintain a healthy vascular system. Various risk factors responsible for hypertension and diabetes may disrupt this homeostasis, leading to decreased bioavailability and/or bioactivity of NO, which potentiates endothelial dysfunction. Plasma NO is a useful indicator of NO homeostasis and vascular endothelial function. Since endothelial function plays a key role in the development and progression of diseases like diabetes and hypertension, we sought to investigate the NO profile in patients having diabetes and hypertension and determine the relationship of NO turnover with the disease. For this purpose, three groups were studied for the NO production. The first group consisted of 74 hypertensive patients, the second group consisted of 72 diabetic patients and the third group consisted of 60 healthy controls. Nitrate synthase activity was evaluated by measuring nitrate level using an automated sample injector connected to an automated NO detector - Ion liquid chromatograph. The plasma concentration of NO was found to be significantly lower in both essential hypertensive patients and diabetic patients without complications as compared to the healthy controls (P < 0.05). This data confirms that different factors like hyperglycemia and blood pressure are seen to have immense influence on NO production.


Applying Persuasive Design Techniques to Influence Data-Entry Behaviors in Primary Care: Repeated Measures Evaluation Using Statistical Process Control.
Persuasive design is an approach that seeks to change the behaviors of users. In primary care, clinician behaviors and attitudes are important precursors to structured data entry, and there is an impact on overall data quality. We hypothesized that persuasive design changes data-entry behaviors in clinicians and thus improves data quality. The objective of this study was to use persuasive design principles to change clinician data-entry behaviors in a primary care environment and to increase data quality of data held in a family health team's reporting system. We used the persuasive systems design framework to describe the persuasion context. Afterward, we designed and implemented new features into a summary screen that leveraged several persuasive design principles. We tested the influence of the new features by measuring its impact on 3 data quality measures (same-day entry, record completeness, and data validity). We also measured the impacts of the new features with a paired pre-post t test and generated XmR charts to contextualize the results. Survey responses were also collected from users. A total of 53 users used the updated system that incorporated the new features over the course of 8 weeks. Based on a pre-post analysis, the new summary screen successfully encouraged users to enter more of their data on the same day as their encounter. On average, the percentage of same-day entries rose by 10.3% for each user (P<.001). During the first month of the postimplementation period, users compensated by sacrificing aspects of data completeness before returning to normal in the second month. Improvements to record validity were marginal over the study period (P=.05). Statistical process control techniques allowed us to study the XmR charts to contextualize our results and understand trends throughout the study period. By conducting a detailed systems analysis and introducing new persuasive design elements into a data-entry system, we demonstrated that it was possible to change data-entry behavior and influence data quality in a reporting system. The results show that using persuasive design concepts may be effective in influencing data-entry behaviors in clinicians. There may be opportunities to continue improving this approach, and further work is required to perfect and test additional designs. Persuasive design is a viable approach to encourage clinician user change and could support better data capture in the field of medical informatics.


Bubble sets: revealing set relations with isocontours over existing visualizations.
While many data sets contain multiple relationships, depicting more than one data relationship within a single visualization is challenging. We introduce Bubble Sets as a visualization technique for data that has both a primary data relation with a semantically significant spatial organization and a significant set membership relation in which members of the same set are not necessarily adjacent in the primary layout. In order to maintain the spatial rights of the primary data relation, we avoid layout adjustment techniques that improve set cluster continuity and density. Instead, we use a continuous, possibly concave, isocontour to delineate set membership, without disrupting the primary layout. Optimizations minimize cluster overlap and provide for calculation of the isocontours at interactive speeds. Case studies show how this technique can be used to indicate multiple sets on a variety of common visualizations.


Delays contributing to the development and repair of obstetric fistula in northern Tanzania.
Delays in receiving obstetric care during labor contribute to high rates of maternal morbidity in sub-Saharan Africa. This exploratory study was conducted to identify important delays experienced during the development and subsequent repair of obstetric fistula in northern Tanzania. Sixty women presenting to a tertiary hospital with obstetric fistula completed structured surveys about the birth experience that led to fistula development and their experiences seeking surgical repair. A subset of 30 provided qualitative accounts. Clinical data were collected postsurgery. Data were analyzed according to a four-delay model, with iterative analysis allowing for triangulation of all sources. During the index pregnancy, women labored for a median of 48 h. Most women (53/60; 88.3%) delivered in a facility but labored for a median of 12.4 h before deciding to seek care (Delay 1). Women spent a median of 1.25 h traveling to a facility (Delay 2). After presenting to care, 15/51 (29.4%) waited at least an hour to see a medical provider, and 35/53 (66.0%) required transfer to another facility (Delay 3). Women lived with fistula for a median of 10 years (Delay 4). Qualitative data provided context and a deeper understanding of the factors contributing to each delay. Critical delays exist both outside and within the healthcare system that contribute to the development and timely repair of obstetric fistula. Healthcare system strengthening, particularly with regard to emergency obstetric care, is critical to reduce the burden of obstetric fistula in women in Tanzania.


Hyperoxia toxicity after cardiac arrest: What is the evidence?
This review gives an overview of current knowledge on hyperoxia pathophysiology and examines experimental and human evidence of hyperoxia effects after cardiac arrest. Oxygen plays a pivotal role in critical care management as a lifesaving therapy through the compensation of the imbalance between oxygen requirements and supply. However, growing evidence sustains the hypothesis of reactive oxygen species overproduction-mediated toxicity during hyperoxia, thus exacerbating organ failure by various oxidative cellular injuries. In the cardiac arrest context, evidence of hyperoxia effects on outcome is fairly conflicting. Although prospective data are lacking, retrospective studies and meta-analysis suggest that hyperoxia could be associated with an increased mortality. However, data originate from retrospective, heterogeneous and inconsistent studies presenting various biases that are detailed in this review. Therefore, after an original and detailed analysis of all experimental and clinical studies, we herein provide new ideas and concepts that could participate to improve knowledge on oxygen toxicity and help in developing further prospective controlled randomized trials on this topic. Up to now, the strategy recommended by international guidelines on cardiac arrest (i.e., targeting an oxyhemoglobin saturation of 94-98 %) should be applied in order to avoid deleterious hypoxia and potent hyperoxia.


PCNA and Ki-67 labelling indices in pre-irradiated and post-irradiated astrocytomas: a comparative immunohistochemical analysis for evaluation of proliferative activity.
To determine the tumour proliferative activity in a series of archival cerebral astrocytomas and compare proliferating cell nuclear antigen (PCNA) and Ki-67 labelling indices in the primary and recurring neoplasms following therapeutic radiation. Twenty eight cases of pre-irradiated and post-irradiated astrocytomas (ranging from WHO grades I to IV) were stained immunohistochemically using the avidin-biotin horseradish peroxidase technique. Two antibodies, PC10 and MIB-1, were used to establish the proliferating labelling indices, PC10 identifies PCNA and MIB-1 recognises the Ki-67 antigen. Both antibodies showed significantly higher labelling indices in the post-irradiated specimens. However, in general, the Ki-67 indices were lower than those for PCNA. MIB-1 immunoreactivity showed less variation and was more intense than that seen with PC10. The discrepancy between the labelling indices of the pre-irradiated and post-irradiated samples raises questions about the evolution of astrocytomas and the effects of therapeutic intervention. The data may represent genetic alterations, the natural tumour course, and/or the effect of radiation. Although both of the antibodies reflected the state of growth of neoplastic cells in astrocytomas, MIB-1 was more reliable. A simple immunohistochemical method using proliferation markers does have an important role in the future care of patients with astrocytoma.


Low-temperature plasma--a prospective microbicidal tool.
The effects of low-temperature plasma treatment on microorganisms typically related to skin diseases are studied qualitatively by the inhibition of growth and viability assays to evaluate the potential for classifying as a prospective antiseptic agent. A variety of microorganisms enveloping gram- negative and gram-positive bacteria as well as one genus of yeast and fungus each were exposed to plasma in vitro. In a comparative approach, two power supplies, both of which produce high voltage pulses yet at different temporal characteristics, are applied for the growth study. While operation with both devices led to growth inhibition of all microbes, the results indicate a superior antimicrobial efficacy for high voltage pulse lengths in the nanosecond scale. Fluorescence assays reveal the efficacy of nanosecond-pulse driven plasma in reducing germ viability. Furthermore, the technical background for patents related to low-temperature plasma technology in the field of plasma medicine is discussed.


Who should be sent for genetic testing in hereditary colorectal cancer syndromes?
Genetic testing is being adopted increasingly to identify individuals with germline mutations that predispose to hereditary colorectal cancer syndromes. Deciding who to test and for which syndrome is of concern to members of the GI oncology community, molecular geneticists, and genetic counselors. The purpose of this review is to help provide guidelines for testing, given that the results influence syndrome diagnosis and clinical management. Although family history may determine whether testing is appropriate and may direct testing to the most informative family member, evolving clinicopathologic features can identify individual patients who warrant testing. Thus, although the usual absence of clinical premonitory signs in hereditary nonpolyposis colorectal cancer (or Lynch syndrome) adds difficulty to its diagnosis, use of the Amsterdam Criteria and Bethesda Guidelines can prove helpful. In contrast, premonitory stigmata such as pigmentations in Peutz-Jeghers syndrome and the phenotypic features of familial adenomatous polyposis aid significantly in syndrome diagnosis. We conclude that the physician's role in advising DNA testing is no small matter, given that a hereditary cancer syndrome's sequelae may be far reaching. Genetic counselors may be extremely helpful to the practicing gastroenterologist, oncologist, or surgeon; when more specialized knowledge is called for, referral can be made to a medical geneticist and/or a medical genetics clinic.


Different Intestinal Microbial Profile in Over-Weight and Obese Subjects Consuming a Diet with Low Content of Fiber and Antioxidants.
Obesity has been related to an increased risk of multiple diseases in which oxidative stress and inflammation play a role. Gut microbiota has emerged as a mediator in this interaction, providing new mechanistic insights at the interface between fat metabolism dysregulation and obesity development. Our aim was to analyze the interrelationship among obesity, diet, oxidative stress, inflammation and the intestinal microbiota in 68 healthy adults (29.4% normal-weight). Diet was assessed through a food frequency questionnaire and converted into nutrients and dietary compounds using food composition tables. The intestinal microbiota was assessed by quantitative PCR, fecal short chain fatty acids by gas chromatography and serum biomarkers by standard protocols. Higher levels of malondialdehyde (MDA), C reactive protein (CRP), serum leptin, glucose, fat percentage and the intestinal Lactobacillus group were found in the obese people. Cluster analysis of body mass index, fat mass, glucose, LDL/HDL ratio, leptin, MDA and CRP classified the subjects into two groups. The levels of the intestinal Bacteroides-Prevotella-Porphyromonas group were lower in the cluster and linked to a higher pro-oxidant and pro-inflammatory status, whose individuals also had lower intake of fruits, dried fruits, and fish. These results could be useful for designing strategies targeted to obesity prevention.


Connective tissue growth factor regulates interneuron survival and information processing in the olfactory bulb.
Neurogenesis underlies plastic changes in defined neuronal circuits in the postnatal and adult brain. Here we identify connective tissue growth factor (CTGF) as a critical factor in the mouse olfactory bulb (OB) in determining the efficiency of incorporation of postnatally born inhibitory neurons, thus gating the output of glomeruli, the first relay station of olfactory processing in the brain. In the OB, CTGF expression was restricted to prenatally born external tufted cells. CTGF enhanced the proapoptotic activity of glial-derived TGF-β2, decreasing the survival of periglomerular inhibitory neurons. Changes in CTGF expression levels in the OB led to modifications in local neuronal circuitry and olfactory behaviors. We show that the odorant-specific recruitment of distinct glomeruli resulted in enhanced local CTGF expression levels in the activated glomeruli. Collectively our data reveal a molecular mechanism controlling the survival of defined postnatally born neurons, thus adapting neuronal integration to the sensory experiences.


Oral Application of Recombinant Bacillus subtilis Spores to Dogs Results in a Humoral Response against Specific Echinococcus granulosus Paramyosin and Tropomyosin Antigens.
Bacillus subtilis is known as an endospore- and biofilm-forming bacterium with probiotic properties. We have recently developed a method for displaying heterologous proteins on the surface of B. subtilis biofilms by introducing the coding sequences of the protein of interest into the bacterial genome to generate a fusion protein linked to the C terminus of the biofilm matrix protein TasA. Although B. subtilis is a regular component of the gut microflora, we constructed a series of recombinant B. subtilis strains that were tested for their ability to be used to immunize dogs following oral application of the spores. Specifically, we tested recombinant spores of B. subtilis carrying either the fluorescent protein mCherry or else selected antigenic peptides (tropomyosin and paramyosin) from Echinococcus granulosus, a zoonotic intestinal tapeworm of dogs and other carnivores. The application of the recombinant B. subtilis spores led to the colonization of the gut with recombinant B. subtilis but did not cause any adverse effect on the health of the animals. As measured by enzyme-linked immunosorbent assay and immunoblotting, the dogs were able to develop a humoral immune response against mCherry as well as against E. granulosus antigenic peptides. Interestingly, the sera of dogs obtained after immunization with recombinant spores of E. granulosus peptides were able to recognize E. granulosus protoscoleces, which represent the infective form of the head of the tapeworms. These results represent an essential step toward the establishment of B. subtilis as an enteric vaccine agent.


A curvilinear effect of height on reproductive success in human males.
Human male height is associated with mate choice and intra-sexual competition, and therefore potentially with reproductive success. A literature review (n = 18) on the relationship between male height and reproductive success revealed a variety of relationships ranging from negative to curvilinear to positive. Some of the variation in results may stem from methodological issues, such as low power, including men in the sample who have not yet ended their reproductive career, or not controlling for important potential confounders (e.g. education and income). We investigated the associations between height, education, income and the number of surviving children in a large longitudinal sample of men (n = 3,578; Wisconsin Longitudinal Study), who likely had ended their reproductive careers (e.g. > 64 years). There was a curvilinear association between height and number of children, with men of average height attaining the highest reproductive success. This curvilinear relationship remained after controlling for education and income, which were associated with both reproductive success and height. Average height men also married at a younger age than shorter and taller men, and the effect of height diminished after controlling for this association. Thus, average height men partly achieved higher reproductive success by marrying at a younger age. On the basis of our literature review and our data, we conclude that men of average height most likely have higher reproductive success than either short or tall men. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1007/s00265-011-1283-2) contains supplementary material, which is available to authorized users.


Spot urine uric acid to creatinine ratio used in the estimation of uric acid excretion in primary gout.
Uric acid overexcretion in patients with gout is frequently assessed by the measurement of 24 hour urinary uric acid excretion, which is cumbersome with ambulatory patients, and requires accurate timing and complete collection of the specimen. We assessed whether uric acid to creatinine ratio (Uua/Ucr) in spot urine is useful for the estimation of uric acid overexcretion in patients with gout. One hundred thirty male patients with gout and 33 non-gout male control subjects were studied. Early morning urine and/or a portion of 24 h collected urine (24 h urine) were used as spot urine samples. Uric acid overexcreters were defined as those with a 24 h urinary uric acid excretion > or = 1000 mg/day, while uric acid underexcreters were defined as those with uric acid clearance < 6 ml/min. There was a significant relationship between 24 h urinary uric acid excretion and early morning urine Uua/Ucr in patients with gout, while no such relationship was observed in controls. No significant difference in Uua/Ucr was observed between patients with gout and controls, or in Uua/Ucr between gout uric acid overexcreters and underexcreters in early morning urine. A significant difference in this value was observed between the 2 groups in the 24 h urine specimens. Although the diagnostic accuracy of gout uric acid overexcretion was 87.2% using early morning urine and 89.6% using 24 h urine, the sensitivity of gout uric acid overexcretion was only 25.0% when using early morning urine and 25.0% when using 24 h urine, when the cutoff value of Uua/Ucr was 0.63 and 0.64, respectively. Uua/Ucr using spot urine, especially early morning urine, is not an accurate indicator of uric acid overexcretion in patients with gout.


Experimentally validated mathematical model of analyte uptake by permeation passive samplers.
A mathematical model describing the sampling process in a permeation-based passive sampler was developed and evaluated numerically. The model was applied to the Waterloo Membrane Sampler (WMS), which employs a polydimethylsiloxane (PDMS) membrane as a permeation barrier, and an adsorbent as a receiving phase. Samplers of this kind are used for sampling volatile organic compounds (VOC) from air and soil gas. The model predicts the spatio-temporal variation of sorbed and free analyte concentrations within the sampler components (membrane, sorbent bed and dead volume), from which the uptake rate throughout the sampling process can be determined. A gradual decline in the uptake rate during the sampling process is predicted, which is more pronounced when sampling higher concentrations. Decline of the uptake rate can be attributed to diminishing analyte concentration gradient within the membrane, which results from resistance to mass transfer and the development of analyte concentration gradients within the sorbent bed. The effects of changing the sampler component dimensions on the rate of this decline in the uptake rate can be predicted from the model. Performance of the model was evaluated experimentally for sampling of toluene vapors under controlled conditions. The model predictions proved close to the experimental values. The model provides a valuable tool to predict changes in the uptake rate during sampling, to assign suitable exposure times at different analyte concentration levels, and to optimize the dimensions of the sampler in a manner that minimizes these changes during the sampling period.


Tryptophan hydroxylase-1 regulates immune tolerance and inflammation.
Nutrient deprivation based on the loss of essential amino acids by catabolic enzymes in the microenvironment is a critical means to control inflammatory responses and immune tolerance. Here we report the novel finding that Tph-1 (tryptophan hydroxylase-1), a synthase which catalyses the conversion of tryptophan to serotonin and exhausts tryptophan, is a potent regulator of immunity. In models of skin allograft tolerance, tumor growth, and experimental autoimmune encephalomyelitis, Tph-1 deficiency breaks allograft tolerance, induces tumor remission, and intensifies neuroinflammation, respectively. All of these effects of Tph-1 deficiency are independent of its downstream product serotonin. Because mast cells (MCs) appear to be the major source of Tph-1 and restoration of Tph-1 in the MC compartment in vivo compensates for the defect, these experiments introduce a fundamentally new mechanism of MC-mediated immune suppression that broadly impacts multiple arms of immunity.


Digital correlation of ion and optical microscopic images: application to the study of thyroglobulin chemical modification.
A method has been developed in order to digitally correlate ion and optical microscopic images of the same sample areas. Serial cross-sections of human thyroid tissue were analyzed by secondary ion mass microscopy and by light microscopy. The resulting chemical and immunochemical map images were superimposed and correlated by means of a two-pass registration algorithm which allows to correct for geometrical distortions introduced by the ion microscope. Results are presented for the study of thyroglobulin chemical modification in pathological thyroid tissue that demonstrates heterogeneous molecular activity.


Characteristics of stride behavior during treadmill walking and stationary stepping.
Much has been learned about the characteristics of gait in overground and treadmill walking. However, there are many contexts in which overground or treadmill walking might not be possible, such as in home-based physical therapy. In those cases, a surrogate task to index gait behavior would be a valuable tool. Thus, the purpose of this study was to evaluate the stride behavior characteristics of stationary stepping compared with treadmill walking. Healthy young adults (N = 10) preformed two 15-minute tasks: (1) treadmill walking and (2) stationary stepping. Several stride behavior characteristics were recorded, including the number of strides taken, minimum and maximum knee angle, stride interval mean, stride interval standard deviation, and detrended fluctuation analysis (DFA) alpha of the stride interval time series. The results showed that stride behavior was similar between tasks when examined at the group level. However, when individual level analyses were used to examine the reliability of each metric between tasks, poor reliability was observed in most metrics, indicating that stationary stepping may not be an appropriate surrogate task for overground or treadmill walking. These results are discussed in the context of a gait dynamics framework, with attention to task constraints that may have influenced the findings.


Selective trihydroxyazepane NagZ inhibitors increase sensitivity of Pseudomonas aeruginosa to β-lactams.
AmpC β-lactamase confers resistance to β-lactam antibiotics in many Gram negative bacteria. Inducible expression of AmpC requires an N-acetylglucosaminidase termed NagZ. Here we describe the synthesis and characterization of hydroxyazepane inhibitors of NagZ. We find that these inhibitors enhance the susceptibility of clinically relevant Pseudomonas aeruginosa to β-lactams.


Coin rotation task (CRT): a new test of motor dexterity.
Coin-rotation task (CRT), a measure of rapid, coordinated finger movements, was devised as a convenient, easily administered bedside test of motor dexterity; however, very little psychometric data exist regarding this task. The current project was undertaken to (a) provide preliminary normative data, (b) examine the convergent and discriminant validity of the task when compared with other standardized motor measures, and (c) examine the diagnostic accuracy of the CRT. The sample of 86 male participants included 60 controls and 26 patients with unilateral lesions of the left (n = 13) and right (n = 13) hemispheres. The CRT was not significantly correlated with age or education; non-adjusted left- and right-hand normative data are provided. The CRT demonstrated good convergent and divergent validity when compared with other standardized motor measures. The CRT was successful in differentiating control and brain damaged groups with mild motor impairment, and demonstrated an overall classification rate of 84.9%. Levels of sensitivity and specificity of the CRT were comparable with or better than other standardized tests of manual dexterity. The CRT offers a valid, quick, and convenient bedside measure of subtle motor impairment.


A Self-Assembled Organic/Metal Junction for Water Photo-Oxidation.
We report the in situ self-assembly of TTF, TTF•+, and BF4- or PF6- into p-type semiconductors on the surface of Pt microparticles dispersed in water/acetonitrile mixtures. The visible light photoactivation of these self-assemblies leads to water oxidation forming O2 and H+, with an efficiency of 100% with respect to the initial concentration of TTF•+. TTF•+ is then completely reduced to TTF upon photoreduction with water. The Pt microparticles act as floating microelectrodes whose Fermi level is imposed by the different redox species in solution; here predominantly TTF, TTF•+, and HTTF+, which furthermore showed no signs of decomposition in solution.


The conserved AAUAAA hexamer of the poly(A) signal can act alone to trigger a stable decrease in RNA polymerase II transcription velocity.
In vivo the poly(A) signal not only directs 3'-end processing but also controls the rate and extent of transcription. Thus, upon crossing the poly(A) signal RNA polymerase II first pauses and then terminates. We show that the G/U-rich region of the poly(A) signal, although required for termination in vivo, is not required for poly(A)-dependent pausing either in vivo or in vitro. Consistent with this, neither CstF, which recognizes the G/U-rich element, nor the polymerase CTD, which binds CstF, is required for pausing. The only part of the poly(A) signal required to direct the polymerase to pause is the AAUAAA hexamer. The effect of the hexamer on the polymerase is long lasting--in many situations polymerases over 1 kb downstream of the hexamer continue to exhibit delayed progress down the template in vivo. The hexamer is the first part of the poly(A) signal to emerge from the polymerase and may play a role independent of the rest of the poly(A) signal in paving the way for subsequent events such as 3'-end processing and termination of transcription.


Gaze aversion as a cognitive load management strategy in autism spectrum disorder and Williams syndrome.
During face-to-face questioning, typically developing children and adults use gaze aversion (GA), away from their questioner, when thinking. GA increases with question difficulty and improves the accuracy of responses. This is the first study to investigate whether individuals with autism spectrum disorder (ASD; associated with reduced sociability and atypical face gaze) and Williams syndrome (WS; associated with hypersociability and atypical face gaze) use GA to manage cognitive load during face-to-face interactions. Two studies were conducted exploring the typicality of GA during face-to-face questioning in (a) ASD and (b) WS. In Study 1, children with ASD increased their GA as question difficulty increased. In addition, they used most GA when thinking about their responses to questions, mirroring evidence from typically developing children. An important atypicality for participants with ASD was a significantly higher level of GA when listening to interlocutors. In Study 2, participants with WS showed typical patterns of GA in relation to question difficulty and across different points of the interaction. Two different neuro-developmental disorders, both characterized by significant problems with executive control of attention and atypicalities of social interactions, exhibited generally typical patterns of GA. All groups used most GA while thinking about questions, and increased their GA as questions got harder. In addition, children with ASD showed elevated levels of GA while listening to questions, but not while thinking about or making their responses, suggesting that they sometimes fail to see the relevance of attending to visual cues rather than actively avoiding them. Results have important implications for how professionals interpret GA in these populations and for social skills training.


Approaches to Develop Alternative Testing Strategies to Inform Human Health Risk Assessment of Nanomaterials.
The development of alternative testing strategies (ATS) for hazard assessment of new and emerging materials is high on the agenda of scientists, funders, and regulators. The relatively large number of nanomaterials on the market and under development means that an increasing emphasis will be placed on the use of reliable, predictive ATS when assessing their safety. We have provided recommendations as to how ATS development for assessment of nanomaterial hazard may be accelerated. Predefined search terms were used to identify the quantity and distribution of peer-reviewed publications for nanomaterial hazard assessment following inhalation, ingestion, or dermal absorption. A summary of knowledge gaps relating to nanomaterial hazard is provided to identify future research priorities and areas in which a rich data set might exist to allow ATS identification. Consultation with stakeholders (e.g., academia, industry, regulators) was critical to ensure that current expert opinion was reflected. The gap analysis revealed an abundance of studies that assessed the local and systemic impacts of inhaled particles, and so ATS are available for immediate use. Development of ATS for assessment of the dermal toxicity of chemicals is already relatively advanced, and these models should be applied to nanomaterials as relatively few studies have assessed the dermal toxicity of nanomaterials to date. Limited studies have investigated the local and systemic impacts of ingested nanomaterials. If the recommendations for research prioritization proposed are adopted, it is envisioned that a comprehensive battery of ATS can be developed to support the risk assessment process for nanomaterials. Some alternative models are available for immediate implementation, while others require more developmental work to become widely adopted. Case studies are included that can be used to inform the selection of alternative models and end points when assessing the pathogenicity of fibers and mode of action of nanomaterial toxicity.


An in vitro study of the probiotic potential of a bile-salt-hydrolyzing Lactobacillus fermentum strain, and determination of its cholesterol-lowering properties.
This study evaluated the use of a bile-salt-hydrolyzing Lactobacillus fermentum strain as a probiotic with potential hypocholesterolemic properties. The effect of L. fermentum on representative microbial populations and overall metabolic activity of the human intestinal microbiota was investigated using a three-stage continuous culture system. Also, the use of galactooligosaccharides as a prebiotic to enhance growth and/or activity of the Lactobacillus strain was evaluated. Administration of L. fermentum resulted in a decrease in the overall bifidobacterial population (ca. 1 log unit). In the in vitro system, no significant changes were observed in the total bacterial, Lactobacillus, Bacteroides, and clostridial populations through L. fermentum supplementation. Acetate production decreased by 9 to 27%, while the propionate and butyrate concentrations increased considerably (50 to 90% and 52 to 157%, respectively). A general, although lesser, increase in the production of lactate was observed with the administration of the L. fermentum strain. Supplementation of the prebiotic to the culture medium did not cause statistically significant changes in either the numbers or the activity of the microbiota, although an increase in the butyrate production was seen (29 to 39%). Results from this in vitro study suggest that L. fermentum KC5b is a candidate probiotic which may affect cholesterol metabolism. The short-chain fatty acid concentrations, specifically the molar proportion of propionate and/or bile salt deconjugation, are probably the major mechanism involved in the purported cholesterol-lowering properties of this strain.


Drug release from matrix tablets: physiological parameters and the effect of food.
As dissolution plays an important and vital role in the drug-delivery process of oral solid dosage forms, it is, therefore, essential to critically evaluate the parameters that can affect this process. The consumption of food as well as the physiological environment and properties of the gastrointestinal tract, such as its volume and composition of fluid, the fluid hydrodynamics, properties of the intestinal membrane, drug dose and solubility, pKa, diffusion coefficient, permeability and particle size, all affect drug dissolution and absorption rate. There are several dissolution approaches that have been developed to address the conditions as experienced in the in vivo environment, as the traditional dissolution being a quality control method is not biorelevant and as such do not always produce meaningful data. This review also describes the development of a systematic way that differentiates between robust and non-robust formulations by varying the effects of agitation and ionic strength through the use of the automated United States Pharmacopeia type III Bio-Dis apparatus. With the improved understanding of the physiological parameters that can affect the oral bioperformance of dosage forms, strides have, therefore, been made in making dissolution testing methods more biologically based with the view of obtaining more in vitro-in vivo correlations.


Anticipatory brainstem activity predicts neural processing of pain in humans.
Previous neuroimaging studies have shown brain activity during not only the application of noxious stimuli, but also prior to stimulation. The functional significance of the anticipatory response, however, has yet to be explored. Two theoretical responses involve either a decrease or an increase in sensitivity of the nociceptive system. In a functional magnetic resonance imaging (fMRI) study, brainstem responses during anticipation and processing of thermal noxious stimuli were investigated. Twelve healthy subjects were warned prior to and then received noxious stimulation to their left hand. Behavioral data showed a positive correlation between the intensity of anticipation and pain. FMRI data revealed brainstem activation in the PAG during the anticipation period. When correlated with individual anticipation ratings, activation during anticipation included significant clusters within the entorhinal cortex and ventral tegmental area (VTA). During receipt, activation within the brainstem included the PAG, VTA, rostral ventromedial medulla (RVM), and the parabrachial nucleus (PB), all elements of descending pain pathways. Using a backward model approach, we explored the functional significance of the anticipatory neural response for subsequent pain processing. Results of this regression analysis revealed that insula activity during receipt was predicted by activity in both the entorhinal cortex and VTA during anticipation. We suggest that activation in both regions before and during pain may underlie anticipation and subsequent pain modulatory responses, possibly involving the appraisal and control of attention necessary for pain modulation. Together, the results suggest a possible role of brainstem areas in anticipatory mechanisms involved in the maintenance of chronic pain.


Peritoneal recurrence of gastric adenocarcinoma after curative resection.
Death from gastric carcinoma after curative resection is mostly due to recurrence. The most common recurrence is peritoneal dissemination, which represents 33 to 50% of total recurrence after curative gastrectomy. Since survival after peritoneal dissemination is very poor, selection of high-risk patients for further management after resection may contribute to better survival. Based on a prospective database, a total of 1,092 patients with gastric carcinoma who underwent curative resection were included in this study. The incidence of peritoneal recurrence, disease-free interval and survival after peritoneal recurrence were determined. The clinicopathological factors including sex, age, tumor location, size, gross appearance, histological findings, depth of tumor invasion, lymph node status, tumor cell infiltration pattern were analyzed to see if there is any relationship with peritoneal dissemination. The incidence of peritoneal recurrence after curative resection was 13.46%. The mean disease-free interval was 14.2 months and the survival after peritoneal recurrence was 4.9 months in average. Almost all the clinicopathological factors were related with peritoneal recurrence, but only depth of tumor invasion, gross appearance and tumor infiltration pattern are closely associated with peritoneal tumor dissemination after multivariate analyses. Since the prognosis after peritoneal recurrence is so poor, selection of high-risk patients for further management after gastrectomy may be based on these three factors.


The reproductive biology and larvae of the first tadpole-bearing frog, Limnonectes larvaepartus.
Most of the reproductive modes of frogs include an exotrophic tadpole, but a number of taxa have some form of endotrophic development that lacks a feeding tadpole stage. The dicroglossid frog genus Limnonectes ranges from China south into Indonesia. The breeding biologies of the approximately 60 described species display an unusual diversity that range from exotrophic tadpoles to endotrophic development in terrestrial nests. There have been mentions of oviductal production of typical, exotrophic tadpoles in an undescribed species of Limnonectes from Sulawesi, Indonesia. Here we examine newly collected specimens of this species, now described as L. larvaepartus and present the first substantial report on this unique breeding mode. Typical exotrophic tadpoles that are retained to an advanced developmental stage in the oviducts of a female frog are birthed into slow-flowing streams or small, non-flowing pools adjacent to the streams.


Flotation-Restricted Environmental Stimulation Technique: A Proposed Therapy for Improving Performance and Recovery in Special Forces Operators - A Narrative Review.
The flotation-restriction environmental stimulation technique (FR) may have utility as a recovery tool for improving performance in elite competitive athletes and Special Operations Forces Operators (SOs). Studies suggest that FR may ameliorate various neurophysiological disorders and improve performance in recreational and elite athletic populations. We sought to understand whether there is evidence to support the use of FR to enhance physiological and psychological performance parameters in the SO population and to provide postulations as to the mechanisms of action of FR therapy. We performed an online literary search of publications dating from 1982 to 2021 and identified 34 sources addressing the aims, depending on population and condition or conditions, being treated. The reported physiological and psychological benefits of FR range from immediate to lasting 4 months. Overall, eight to twelve FR treatment sessions of from 40 to 90 minutes each may provide variable long-term benefits. The associated synergistic benefits of FR may be attributed to its thermal, chemical, and mechanical effects but deserve further exploration. Based on the current evidence, FR may serve as an effective performance-recovery therapy for improving pain, sleep, and performance measures (e.g., marksmanship and physical performance) in trained, untrained, and healthy adults. Future research focusing on FR as a unimodal recovery intervention is warranted in a specialized group of SOs.


Comparing different methods of assessing body composition in end-stage renal failure.
Accurate measurement of nutritional status in patients with end-stage renal disease is important because of its clear association with prognosis. Total body water (TBW) has additionally been recently recognized as an independent prognostic value because of its relationship with hypertension and cardiac morbidity. The current study was designed to assess the utility of surrogate markers of nutritional state and TBW in patients with end-stage renal disease. Fifty-four patients with renal disease were studied. TBW obtained using the deuterium dilution technique was compared with estimates derived from anthropometric measures of TBW, including 58% body weight, Watson equations, and bioelectrical impedance analysis (BIA). Anthropometrically derived fat-free mass (FFM) was compared with BIA-derived estimates. Total body nitrogen (TBN) measurements were correlated with TBW estimates and BIA-derived resistance. TBW was significantly underestimated by the Watson equation (mean difference, -1.751 L, P = 0.01) and the 58% body weight approximation significantly overestimated it (mean difference, 1.792 L, P = 0.04). The Kushner BIA estimation of TBW did not significantly differ from that of the gold standard determined from D2O dilution (mean difference, -1.221 L, P = 0.12) and was also the method that showed the best agreement with the D2O estimate. However, the limits of agreement were large. Accurate prediction equations for FFM (FFM = -21.768 + 0.001 x ht2 + 6630.669 x 1/R + 0.312 x wt, R2 = 0.95) and TBN (TBN = -668.324 - 3.963 x age + 10.133 x wt + 0. 045 x ht2 + 32141.457 x 1/R, R2 = 0.91) were derived from BIA obtained resistance. The estimation of TBW varies significantly depending on the method of calculation. BIA is the most accurate surrogate marker for the measurement of both TBW and other parameters of body composition.


Global control in Pseudomonas fluorescens mediating antibiotic synthesis and suppression of black root rot of tobacco.
Pseudomonas fluorescens CHA0 colonizes plant roots, produces several secondary metabolites in stationary growth phase, and suppresses a number of plant diseases, including Thielaviopsis basicola-induced black root rot of tobacco. We discovered that mutations in a P. fluorescens gene named gacA (for global antibiotic and cyanide control) pleiotropically block the production of the secondary metabolites 2,4-diacetylphloroglucinol (Phl), HCN, and pyoluteorin. The gacA mutants of strain CHA0 have a drastically reduced ability to suppress black root rot under gnotobiotic conditions, supporting the previous observations that the antibiotic Phl and HCN individually contribute to the suppression of black root rot. The gacA gene is directly followed by a uvrC gene. Double gacA-uvrC mutations render P. fluorescens sensitive to UV irradiation. The gacA-uvrC cluster is homologous to the orf-2 (= uvrY)-uvrC operon of Escherichia coli. The gacA gene specifies a trans-active 24-kDa protein. Sequence data indicate that the GacA protein is a response regulator in the FixJ/DegU family of two-component regulatory systems. Expression of the gacA gene itself was increased in stationary phase. We propose that GacA, perhaps activated by conditions of restricted growth, functions as a global regulator of secondary metabolism in P. fluorescens.


Effect of mental stress on the tone of a medium-sized muscular artery.
We studied the effects of mental stress on the tone of the radial artery, a medium-sized muscular artery. Using an A-mode echo-tracking device coupled to a photoplethysmograph, we took continuous measurements of the radial artery diameter, heart rate and finger arterial pressure in six healthy young volunteers. Under mental stress, the heart rate increased from 63 +/- 4 to 74 +/- 4 beats/min (mean +/- SEM, P < 0.01) and systolic pressure from 123 +/- 5 to 140 +/- 6 mmHg (P < 0.05). No consistent modification in the arterial diameter was observed at this time. The activation of sympathetic nerve activity induced by mental stress does not cause a significant contraction in the radial artery.


Patients' instability, emergency response, and outcomes in the radiology department.
Inpatients may be at risk of cardiopulmonary instability during radiologic testing. Calling the medical emergency team is one rescue intervention that brings a team of critical care providers to the unstable patient. Little is known, however, about patients' instability and activations of the medical emergency team in the radiology department (RD-MET). To describe the cause of activation of the RD-MET for hospitalized patients, temporal attributes of RD-MET involvement, characteristics of RD-MET patients, and characteristics associated with good and poor outcomes after RD-MET activation. Retrospective pilot study of RD-MET calls for 64 inpatients in a tertiary care hospital during 2009. Reasons for RD-MET activation were 39% neurological, 38% cardiac, and 22% respiratory, and nearly half (42%) occurred during a computed tomography scan. Most RD-MET calls were made between 10 am and noon. RD-MET patients had a mean age of 61 (SD, 19) years; 52% were female, and 89% were white. Admitting diagnoses were most commonly neurological (20%), cardiovascular (16%), and abdominal (16%). The most common comorbid conditions were chronic obstructive pulmonary disease (23%) and diabetes (20%). Half of RD-MET inpatients were from a general care unit, and 56% required preexisting oxygen support. After RD-MET involvement, 61% of patients required a higher level of care; 3% died during the MET intervention, and 19% died later in hospitalization. Patients with preexisting comorbid conditions were more likely to have poor outcomes after the RD-MET intervention (P = .001). RD-MET patients with comorbid conditions, from a general care unit, and at risk for neurological deterioration arrive in the radiology department with potentially underestimated support needs. Greater support in specific time frames and locations may be warranted to improve outcomes.


Cross-resistance to tumour promoters in human cancer cell lines resistant to adriamycin or cisplatin.
The growth inhibitory effect of tumour promoters on human leukaemia and lung cancer cell lines was examined using the [3-(4,5 dimethylthiazol)-2, 5-diphenyl-tetrazolium bromide (MTT) assay. The four cell lines used were the K562 human leukaemia cell line, its adriamycin (ADM)-resistant subline (K562/ADM), which shows the mdr phenotype, PC-9 (a human lung adenocarcinoma cell line) and its cisplatin (CDDP)-resistant subline (PC-9/CDDP), which does not show the mdr phenotype. Phorbol 12-tetradecanoate-13-acetate (TPA) and the TPA-type tumour promoters, aplysiatoxin and debromoaplysiatoxin, inhibited the growth of the two parental cell lines, K562 and PC-9. The non-TPA-type tumour promoter, okadaic acid, also inhibited the growth of the two parental cell lines in a dose-dependent manner. TPA-type and okadaic acid inhibited the growth of K562/ADM more weakly than that of K562, and showed no growth inhibition in PC-9/CDDP. Anhydrodebromoaplysiatoxin, an inactive derivative of the TPA-type tumour promoter, could suppress the growth of K562 and K562/ADM only at high concentration (more than 50 pM) and it showed similar growth inhibitory effects on the two cell lines. Okadaic acid tetramethyl ether, the inactive form of the non-TPA-type tumour promoter did not inhibit the growth of any of the cell lines. The growth inhibitory effect of these compounds was well correlated with their tumour-promoting activity. A study of the accumulation of okadaic acid revealed that the amount of 3H-okadaic acid in K562/ADM and PC-9/CDDP was similar to that in their parental cells indicating that cross-resistance to this tumour promoter in the drug-resistant cell lines is not due to a difference in the amount of drug accumulated in sensitive and resistant cells. These results suggest the presence of another common mechanism for resistance to ADM and CDDP as well as to TPA- or non-TPA-type tumour promoters.


Dosage changes of MED13L further delineate its role in congenital heart defects and intellectual disability.
A chromosomal balanced translocation disrupting the MED13L (Mediator complex subunit13-like) gene, encoding a subunit of the Mediator complex, was previously associated with transposition of the great arteries (TGA) and intellectual disability (ID), and led to the identification of missense mutations in three patients with isolated TGA. Recently, a homozygous missense mutation in MED13L was found in two siblings with non-syndromic ID from a consanguineous family. Here, we describe for the first time, three patients with copy number changes affecting MED13L and delineate a recognizable MED13L haploinsufficiency syndrome. Using high resolution molecular karyotyping, we identified two intragenic de novo frameshift deletions, likely resulting in haploinsufficiency, in two patients with a similar phenotype of hypotonia, moderate ID, conotruncal heart defect and facial anomalies. In both, Sanger sequencing of MED13L did not reveal any pathogenic mutation and exome sequencing in one patient showed no evidence for a non-allelic second hit. A further patient with hypotonia, learning difficulties and perimembranous VSD showed a 1 Mb de novo triplication in 12q24.2, including MED13L and MAP1LC3B2. Our findings show that MED13L haploinsufficiency in contrast to the previously observed missense mutations cause a distinct syndromic phenotype. Additionally, a MED13L copy number gain results in a milder phenotype. The clinical features suggesting a neurocristopathy may be explained by animal model studies indicating involvement of the Mediator complex subunit 13 in neural crest induction.


Small Nucleolar RNA Score: An Assay to Detect Formalin-Overfixed Tissue.
Although there are millions of formalin-fixed paraffin-embedded (FFPE) tissue blocks potentially available for scientific research, many are of questionable quality, partly due to unknown fixation conditions. We analyzed FFPE tissue biospecimens as part of the NCI Biospecimen Preanalytical Variables (BPV) program to identify microRNA (miRNA) markers for fixation time. miRNA was extracted from kidney and ovary tumor FFPE blocks (19 patients, cold ischemia ≤2 hours) with 6, 12, 24, and 72 hours fixation times, then analyzed using the WaferGen SmartChip platform (miRNA chip with 1036 miRNA targets). For fixation time, principal component analysis of miRNA chip expression data separated 72 hours fixed samples from 6 to 24 hours fixed samples. A set of small nuclear RNA (snRNA) targets was identified that best determines fixation time and was validated using a second independent cohort of seven different tissue types. A customized assay was then developed, based on a set of 24 miRNA and snRNA targets, and a simple "snoRNA score" defined. This score detects FFPE tissue samples with fixation for 72 hours or more, with 79% sensitivity and 80% specificity. It can therefore be used to assess the fitness-for-purpose of FFPE samples for DNA or RNA-based research or clinical assays, which are known to be of limited robustness to formalin overfixation.


Multiple hemopoietic lineages are found after stimulation of mouse bone marrow precursor cells with interleukin 3.
When the murine T-lymphocyte clone L2 is stimulated with concanavalin A, it secretes at least two distinct factors that affect hemopoietic precursor cells, interleukin 3 (IL3) and granulocyte/macrophage colony-stimulating factor (GM-CSF). IL3 accounts for approximately 10% of the colony-stimulating activity in L2-cell-conditioned medium. The IL3 secreted by L2 cells is similar antigenically to the IL3 secreted by WEHI-3 cells. Like the IL3 from WEHI-3 cells, IL3 secreted by L2 cells does not bind to DEAE Sephacel and can be separated from the L2-cell GM-CSF, which does bind to DEAE. By assessment of the functional, morphologic, surface phenotypic, and cytochemical characteristics of bone marrow cells 6 days after stimulation with IL3 in liquid culture, four hemopoietic lineages were found, including macrophage, neutrophilic granulocyte, megakaryocyte, and basophil/mast cell. In addition, when bone marrow cells were stimulated with IL3 in semisolid medium, several types of colonies were found, including mixed colonies containing macrophage, megakaryocyte, and granulocyte lineages.


Sarcomatoid Type Primary Pericardial Mesothelioma with a Long-term Survival after the Onset of Cardiac Tamponade.
Primary pericardial malignant mesothelioma is a very rare clinical entity and its prognosis is very poor. We herein report a 67-year-old man who presented with pericardial mesothelioma that was diagnosed 21 months after the onset of cardiac tamponade as the initial manifestation. Despite undergoing pericardiocentesis and surgical pericardial fenestration at the onset of cardiac tamponade, we were unable to make a conclusive diagnosis of mesothelioma based on the cytological and histological findings. This unusual case had a relatively long progression-free period without treatment before the appearance of pleural tumors that showed the histological features of malignant sarcomatoid mesothelioma.


Bacterial superantigens promote acute nasopharyngeal infection by Streptococcus pyogenes in a human MHC Class II-dependent manner.
Establishing the genetic determinants of niche adaptation by microbial pathogens to specific hosts is important for the management and control of infectious disease. Streptococcus pyogenes is a globally prominent human-specific bacterial pathogen that secretes superantigens (SAgs) as 'trademark' virulence factors. SAgs function to force the activation of T lymphocytes through direct binding to lateral surfaces of T cell receptors and class II major histocompatibility complex (MHC-II) molecules. S. pyogenes invariably encodes multiple SAgs, often within putative mobile genetic elements, and although SAgs are documented virulence factors for diseases such as scarlet fever and the streptococcal toxic shock syndrome (STSS), how these exotoxins contribute to the fitness and evolution of S. pyogenes is unknown. Here we show that acute infection in the nasopharynx is dependent upon both bacterial SAgs and host MHC-II molecules. S. pyogenes was rapidly cleared from the nasal cavity of wild-type C57BL/6 (B6) mice, whereas infection was enhanced up to ∼10,000-fold in B6 mice that express human MHC-II. This phenotype required the SpeA superantigen, and vaccination with an MHC -II binding mutant toxoid of SpeA dramatically inhibited infection. Our findings indicate that streptococcal SAgs are critical for the establishment of nasopharyngeal infection, thus providing an explanation as to why S. pyogenes produces these potent toxins. This work also highlights that SAg redundancy exists to avoid host anti-SAg humoral immune responses and to potentially overcome host MHC-II polymorphisms.


Malaria impact of large dams in sub-Saharan Africa: maps, estimates and predictions.
While there is growing recognition of the malaria impacts of large dams in sub-Saharan Africa, the cumulative malaria impact of reservoirs associated with current and future dam developments has not been quantified. The objective of this study was to estimate the current and predict the future impact of large dams on malaria in different eco-epidemiological settings across sub-Saharan Africa. The locations of 1268 existing and 78 planned large dams in sub-Saharan Africa were mapped against the malaria stability index (stable, unstable and no malaria). The Plasmodium falciparum infection rate (PfIR) was determined for populations at different distances (<1, 1-2, 2-5, 5-9 km) from the associated reservoirs using the Malaria Atlas Project (MAP) and WorldPop databases. Results derived from MAP were verified by comparison with the results of detailed epidemiological studies conducted at 11 dams. Of the 1268 existing dams, 723 are located in malarious areas. Currently, about 15 million people live in close proximity (<5 km) to the reservoirs associated with these dams. A total of 1.1 million malaria cases annually are associated with them: 919,000 cases due to the presence of 416 dams in areas of unstable transmission and 204,000 cases due to the presence of 307 dams in areas of stable transmission. Of the 78 planned dams, 60 will be located in malarious areas and these will create an additional 56,000 cases annually. The variation in annual PfIR in communities as a function of distance from reservoirs was statistically significant in areas of unstable transmission but not in areas of stable transmission. In sub-Saharan Africa, dams contribute significantly to malaria risk particularly in areas of unstable transmission. Additional malaria control measures are thus required to reduce the impact of dams on malaria.


Delay of disease development in transgenic plants that express the tobacco mosaic virus coat protein gene.
A chimeric gene containing a cloned cDNA of the coat protein (CP) gene of tobacco mosaic virus (TMV) was introduced into tobacco cells on a Ti plasmid of Agrobacterium tumefaciens from which tumor inducing genes had been removed. Plants regenerated from transformed cells expressed TMV mRNA and CP as a nuclear trait. Seedlings from self-fertilized transgenic plants were inoculated with TMV and observed for development of disease symptoms. The seedlings that expressed the CP gene were delayed in symptom development and 10 to 60 percent of the transgenic plants failed to develop symptoms for the duration of the experiments. Increasing the concentration of TMV in the inoculum shortened the delay in appearance of symptoms. The results of these experiments indicate that plants can be genetically transformed for resistance to virus disease development.