Essential role of the E3 ubiquitin ligase Cbl-b in T cell anergy induction.
Antigen-specific immunotolerance limits the expansion of self-reactive T cells involved in autoimmune diseases. Here, we show that the E3 ubiquitin ligase Cbl-b is upregulated in T cells after tolerizing signals. Loss of Cbl-b in mice results in impaired induction of T cell tolerance both in vitro and in vivo. Importantly, rechallenge of Cbl-b mutant mice with the tolerizing antigen results in massive lethality. Moreover, ablation of Cbl-b resulted in exacerbated autoimmunity. Mechanistically, loss of Cbl-b rescues reduced calcium mobilization of anergic T cells, which was attributed to Cbl-b-mediated regulation of PLCgamma-1 phosphorylation. Our results show a critical role for Cbl-b in the regulation of peripheral tolerance and anergy of T cells.


Detection of Tip-Burn Stress on Lettuce Grown in an Indoor Environment Using Deep Learning Algorithms.
Lettuce grown in indoor farms under fully artificial light is susceptible to a physiological disorder known as tip-burn. A vital factor that controls plant growth in indoor farms is the ability to adjust the growing environment to promote faster crop growth. However, this rapid growth process exacerbates the tip-burn problem, especially for lettuce. This paper presents an automated detection of tip-burn lettuce grown indoors using a deep-learning algorithm based on a one-stage object detector. The tip-burn lettuce images were captured under various light and indoor background conditions (under white, red, and blue LEDs). After augmentation, a total of 2333 images were generated and used for training using three different one-stage detectors, namely, CenterNet, YOLOv4, and YOLOv5. In the training dataset, all the models exhibited a mean average precision (mAP) greater than 80% except for YOLOv4. The most accurate model for detecting tip-burns was YOLOv5, which had the highest mAP of 82.8%. The performance of the trained models was also evaluated on the images taken under different indoor farm light settings, including white, red, and blue LEDs. Again, YOLOv5 was significantly better than CenterNet and YOLOv4. Therefore, detecting tip-burn on lettuce grown in indoor farms under different lighting conditions can be recognized by using deep-learning algorithms with a reliable overall accuracy. Early detection of tip-burn can help growers readjust the lighting and controlled environment parameters to increase the freshness of lettuce grown in plant factories.


Structure of IL-22 bound to its high-affinity IL-22R1 chain.
IL-22 is an IL-10 family cytokine that initiates innate immune responses against bacterial pathogens and contributes to immune disease. IL-22 biological activity is initiated by binding to a cell-surface complex composed of IL-22R1 and IL-10R2 receptor chains and further regulated by interactions with a soluble binding protein, IL-22BP, which shares sequence similarity with an extracellular region of IL-22R1 (sIL-22R1). IL-22R1 also pairs with the IL-20R2 chain to induce IL-20 and IL-24 signaling. To define the molecular basis of these diverse interactions, we have determined the structure of the IL-22/sIL-22R1 complex. The structure, combined with homology modeling and surface plasmon resonance studies, defines the molecular basis for the distinct affinities and specificities of IL-22 and IL-10 receptor chains that regulate cellular targeting and signal transduction to elicit effective immune responses.


Toxicity reference values for methylmercury effects on avian reproduction: Critical review and analysis.
Effects of mercury (Hg) on birds have been studied extensively and with increasing frequency in recent years. The authors conducted a comprehensive review of methylmercury (MeHg) effects on bird reproduction, evaluating laboratory and field studies in which observed effects could be attributed primarily to Hg. The review focuses on exposures via diet and maternal transfer in which observed effects (or lack thereof) were reported relative to Hg concentrations in diet, eggs, or adult blood. Applicable data were identified for 23 species. From this data set, the authors identified ranges of toxicity reference values suitable for risk-assessment applications. Typical ranges of Hg effect thresholds are approximately 0.2 mg/kg to >1.4 mg/kg in diet, 0.05 mg/kg/d to 0.5 mg/kg/d on a dose basis, 0.6 mg/kg to 2.7 mg/kg in eggs, and 2.1 mg/kg to >6.7 mg/kg in parental blood (all concentrations on a wet wt basis). For Hg in avian blood, the review represents the first broad compilation of relevant toxicity data. For dietary exposures, the current data support TRVs that are greater than older, commonly used TRVs. The older diet-based TRVs incorporate conservative assumptions and uncertainty factors that are no longer justified, although they generally were appropriate when originally derived, because of past data limitations. The egg-based TRVs identified from the review are more similar to other previously derived TRVs but have been updated to incorporate new information from recent studies. While important research needs remain, a key recommendation is that species not yet tested for MeHg toxicity should be evaluated using toxicity data from tested species with similar body weights. Environ Toxicol Chem 2017;36:294-319. © 2016 SETAC.


[Evolution of ideas on microscopic colitis].
The literature review gives the present-day views of the definition, etiology, pathogenesis, diagnosis, and treatment of microscopic colitis (MC). In the present view, MC is an inflammatory bowel disease of unknown etiology, which is characterized by chronic watery diarrhea, no macroscopic signs of large bowel involvement in the presence of specific pathomorphological changes. There are two major forms of MC, which are similar in its clinical picture, yet, heterogeneous in histological criteria: collagenous colitis (CC) and lymphocytic colitis (LC). As of now, the prevalence of MC is about 100 cases per 100,000 population, which is similar with that in other inflammatory bowel diseases, such as ulcerative colitis and Crohn's disease. MC generally prevails in women aged over 50 years. The etiology and pathogenesis of MC have not fully investigated. Watery diarrhea is as a predominant pathognomonic symptom in all the patients with MC. The major histological criterion for the diagnosis of CC is subepithelial collagen lining thickening (more than 10 pm) and that for LC is higher intraepithelial lymphocyte counts (more than 20 intraepithelial lymphocytes/100 epitheliocytes). The topical glucocorticosteroid budesonide is currently the only agent, the efficacy of which has been proven in both inducing and maintaining remission in patients with MC in many clinical trials.


Synthesis and properties of near infrared-absorbing magnetic-optical nanopins.
Novel near infrared-absorbing iron oxide-gold core-shell nanoparticles in pin shapes were synthesized. The nanopins are superparamagnetic, with 35-fold better surface enhanced Raman scattering activities than the conventional core-shell nanospheres and 50-fold greater photothermal properties than solid gold nanorods. The nanoparticles will have important impact on medical imaging, molecular diagnostics and disease treatment.


Prevalence and correlates of vaccine attitudes and behaviors in a cohort of low-income mothers.
The US is facing a rise in vaccine hesitancy, delay, and refusal, though little is known about these outcomes in socio-economically disadvantaged populations. This study examines the prevalence and correlates of vaccine attitudes and behaviors in a diverse cohort of low-income mothers receiving home visiting services. Survey data were collected from 813 recipients of evidence-based home visiting services in Wisconsin from 2013 to 2018. Analyses were performed to describe outcome measures of vaccine attitudes and self-reported completion, and multivariate regressions were used to test associations between vaccine-related outcomes and hypothesized correlates. Most women (94%) reported their children were up to date on vaccines; 14.3% reported having ever delayed vaccination. A small minority disagreed that vaccines are important (5.0%), effective (5.4%), and safe (6.2%), though a larger proportion responded ambivalently (10.9%-21.9%). Participants with greater trust in health care providers reported more positive overall vaccine attitudes (B = 0.24; 95% CI = 0.17, 0.31), a lower likelihood of vaccine delay (OR = 0.57; 95% CI = 0.46, 0.73), and a greater likelihood of being up to date on vaccines (OR = 1.79, 95% CI = 1.30, 2.44). Women with greater trust in a home visitor also rated vaccines more positively (B = 0.09; 95% CI = 0.02, 0.15), and women who reported better mental health were more likely to report their children were up to date (OR = 1.05; 95% CI = 1.02, 1.09). Compared to non-Hispanic whites, American Indians and non-Hispanic blacks had poorer vaccine-related outcomes. More research on vaccine attitudes and behaviors among higher-risk populations is needed to develop tailored strategies aimed at addressing vaccine hesitancy and underimmunization.


[The coverage of science in television news programs in Brazil and Colombia: a comparative study of media constructs].
This study analyzes and compares the science and technology coverage in Brazil's main television news program (Jornal Nacional) and its Colombian counterpart (Noticias Caracol). Using content analysis, we investigated a corpus of news stories broadcast from April 2009 to March 2010. We found that Jornal Nacional presented over twice as many reports on science and technology as Noticias Caracol, and that its levels of reporting remained fairly stable throughout the year. The Brazilian reports were also longer, were featured more prominently, and used more visual resources. Even so, some similarities were found: news about health and medicine was most frequent; the reports focused primarily on announcing new research; scientists were the main sources cited; and national research was prioritized.


Dissecting the determinants of malaria chronicity: why within-host models struggle to reproduce infection dynamics.
The duration of infection is fundamental to the epidemiological behaviour of any infectious disease, but remains one of the most poorly understood aspects of malaria. In endemic areas, the malaria parasite Plasmodium falciparum can cause both acute, severe infections and asymptomatic, chronic infections through its interaction with the host immune system. Frequent superinfection and massive parasite genetic diversity make it extremely difficult to accurately measure the distribution of infection lengths, complicating the estimation of basic epidemiological parameters and the prediction of the impact of interventions. Mathematical models have qualitatively reproduced parasite dynamics early during infection, but reproducing long-lived chronic infections remains much more challenging. Here, we construct a model of infection dynamics to examine the consequences of common biological assumptions for the generation of chronicity and the impact of co-infection. We find that although a combination of host and parasite heterogeneities are capable of generating chronic infections, they do so only under restricted parameter choices. Furthermore, under biologically plausible assumptions, co-infection of parasite genotypes can alter the course of infection of both the resident and co-infecting strain in complex non-intuitive ways. We outline the most important puzzles for within-host models of malaria arising from our analysis, and their implications for malaria epidemiology and control.


Risk factors affecting children of mothers with borderline personality disorder: Comment on Stepp, Whalen, Pilkonis, Hipwell, and Levine (2011).
Comments on an article Children of mothers with borderline personality disorder: Identifying parenting behaviors as potential targets for intervention by Stepp, Whalen, Pilkonis, Hipwell, and Levine (see record 2011-05873-001). Despite both a limited empirical literature and the muddy problem of borderline personality disorder (BPD) classification, the authors have significantly clarified the risk factors relevant to children of mothers with BPD and have outlined important and reasonable skill-based interventions likely to mitigate these risks. Not only have they sensibly connected the dots among the extant studies (which included very different samples, ages of children, criterion behaviors, etc.), they have identified the few key points of agreement across studies and translated these into a blueprint for early intervention. This commentary (a) further elaborates on some of the parameters of BPD that make risk assessment for children of mothers with BPD complicated, including both diagnostic heterogeneity and myriad other problems that reduce the specificity of BPD as a classification tool for these purposes and (b) identifies direct, indirect, and general mechanisms of transmission of difficulties from a mother (or parent or caregiver) with BPD to her child.


Microbiological quality of frozen precooked and peeled shrimp from South-East Asia and from the North Sea.
The microbiological quality of frozen precooked and peeled shrimp is largely dependent upon the method of processing. The investigations were carried out in order to study the difference between the microbiological quality of shrimp from South-East Asia and from the North Sea and to determine whether both can meet draft FAO/WHO and ICMSF-specifications; 100 samples of frozen precooked and peeled shrimp from South-East Asia and 50 samples of shrimp from the North Sea were examined. In 45% of the shrimp samples from South-East Asia and in 78% of the samples of North Sea shrimp the number of aerobic mesophiles was over 10(6) per g. The number of Staph. aureus was higher in the samples of shrimp from South-East Asia than in those from the North Sea: 34% of the samples from South-East Asia had over 2 x 10(3) Staph. aureus per g., whereas only 6% of the samples of North Sea shrimp exceeded this number. Gram-positive mesophilic cocci were predominant in shrimp from South-East Asia. Gram-negative psychrophilic bacteria were predominant in North Sea shrimp. This difference in bacterial flora is caused by a difference in processing. The number of Enterobacteriaceae was usually low (less than 2 x 10(2) per g) in both kinds of shrimp. In three lots of South-East Asian shrimp Salmonella was detected. Vibrio parahaemolyticus was not found. On the basis of draft FAO/WHO endproduct specifications all lots of both South-East Asian and North Sea shrimp would have been rejected. On the basis of ICMSF specifications the rejection amounted to 90 and 80% of the lots, for the two types of shrimp respectively. Preservatives were not detected in shrimp from South-East Asia, whereas benzoic acid was present in all samples of North Sea shrimp. It was concluded that the ammonia content is a valuable objective quality index only in case of North Sea shrimp.


Plumisclerin A, a diterpene with a new skeleton from the soft coral Plumigorgia terminosclera.
A novel compound, named plumisclerin A (1), was isolated from samples of the soft coral Plumigorgia terminosclera collected at Mayotte Island. The compound possesses the novel plumisclerane carbon skeleton, including a tricyclo[4,3,1,0(1,5)]decane ring. Its structure and relative stereochemistry were elucidated by extensive spectroscopic analysis, including HREIMS, COSY, HSQC, HMBC, TOCSY, and NOESY experiments. In addition, the novel compound displayed in vitro cytotoxicity against selected cancer cell lines.


Bridging the Gap Between Consumers' Medication Questions and Trusted Answers.
This paper addresses the task of answering consumer health questions about medications. To better understand the challenge and needs in terms of methods and resources, we first introduce a gold standard corpus for Medication Question Answering created using real consumer questions. The gold standard (https://github.com/abachaa/Medication_QA_MedInfo2019) consists of six hundred and seventy-four question-answer pairs with annotations of the question focus and type and the answer source. We first present the manual annotation and answering process. In the second part of this paper, we test the performance of recurrent and convolutional neural networks in question type identification and focus recognition. Finally, we discuss the research insights from both the dataset creation process and our experiments. This study provides new resources and experiments on answering consumers' medication questions and discusses the limitations and directions for future research efforts.


Relationship between the architecture of zinc coordination and zinc binding affinity in proteins--insights into zinc regulation.
Zinc proteins are an integral component of the proteome of all domains of life. Zn(II), one of the most widespread transition elements, serves multiple functions in proteins, such as a catalytic co-factor, a structural center and a signaling component. The mechanism by which proteins associate with and dissociate from Zn(II) and the factors that modulate their affinity and stability remain incompletely understood. In this article, we aim to address how zinc binding sites present in proteins differ in their architecture and how their structural arrangement is associated with protein function, thermodynamic and kinetic stability, reactivity, as well as zinc-dependent regulation. Here, we emphasize that the concentration-dependent functionality of the interprotein zinc binding site may serve as another factor regulating the relationship between cellular Zn(II) availability and protein function.


Synergism of fused bicyclic 2-aminothiazolyl compounds with polymyxin B against Klebsiella pneumoniae.
A series of fused bicyclic 2-aminothiazolyl compounds were synthesized and evaluated for their synergistic effects with polymyxin B (PB) against Klebsiella pneumoniae (SIPI-KPN-1712). Some of the synthesized compounds exhibited synergistic activity. When 4 μg ml-1 compound B1 was combined with PB, it showed potent antibacterial activity, achieving 64-fold reduction of the MIC of PB. Furthermore, compound B1 showed prominent synergistic efficacy in both concentration gradient and time-kill curves in vitro. In addition, B1 combined with PB also exhibited synergistic and partial synergistic effect against E. coli (ATCC25922 and its clinical isolates), Acinetobacter baumannii (ATCC19606 and its clinical isolates), and Pseudomonas aeruginosa (Pae-1399).


Study of the stepwise deprotonation reactions of glyphosate and the corresponding pKa values in aqueous solution.
Glyphosate (N-(phosphonomethyl)glycine) (Gph) is a herbicide that is broadly used in several countries. Its application to eliminate weeds may have the undesired effect of diminishing the metallic cations found in the soil (e.g., Ni(2+) and Zn(2+)), due to a complexation reaction that depends on the soil's pH. To better understand the molecular structures of glyphosate that are involved in such a complexation reaction, we have studied all possible glyphosate conformations in aqueous solution that may be involved in deprotonation reactions in the pH range from 2 to 11 using the polarizable continuum method (PCM). We have also compared direct (or absolute) methods to calculate pKa values, the cluster-continuum model and the proton-exchange scheme, using different thermodynamic cycles. The best result was achieved when using a proton-exchange scheme, which was able to properly reproduce three glyphosate experimental pKa values predicted for the glyphosate structures and conformations previously determined.


Physical therapies for prevention of urinary and faecal incontinence in adults.
Physical therapies, particularly pelvic floor muscle training, have been recommended for prevention of incontinence associated with childbearing, and prostate surgery. To assess the effectiveness of physical therapies in preventing incontinence in adults. The Cochrane Incontinence Group trials register was searched to September 2001. Trials were also sought from the Reference Lists of relevant articles and from experts in the field. Randomised and quasi-randomised trials in adults without incontinence symptoms that compared a physical therapy with no treatment, or any other treatment to prevent incontinence. Two reviewers independently assessed eligibility, trial quality and extracted data. Data were processed as described in the Cochrane Collaboration Handbook. Two trials in men (155 men) and 13 trials in women (4661 women) were included. As most trials recruited regardless of continence status, and there was much heterogeneity, only a qualitative synthesis was undertaken. Three of seven trials in childbearing women reported less urinary incontinence after pelvic floor muscle training compared to control treatment three months postpartum. Two trials selected women at higher risk of postnatal incontinence. The third used an intensive training programme. Four trials did not find any difference between the groups at the primary endpoint. Two trials compared pre-prostate surgery pelvic floor muscle training with control treatment, and no difference in the occurrence of postoperative urinary incontinence was reported between the groups. There is insufficient evidence to determine whether physical therapies can prevent incontinence in childbearing women, or men following prostate surgery. Further, better quality research is needed.


Phase 1 healthy volunteer willingness to participate and enrollment preferences.
Healthy volunteers in phase 1 clinical trials contribute to the development of safe drugs and other biologics and accept risks and burdens without anticipated health benefits from participation. Although emerging data have shown that healthy volunteers are influenced by risk, some still worry that financial incentives lead them to take on unreasonable risk. Yet little is known about healthy volunteers' preferences and how they make choices about enrolling in research studies. We surveyed 654 healthy volunteers at the end of their participation in a phase 1 Pfizer trial in the United States, Belgium, and Singapore to examine their reported willingness to enroll in studies of different types, with various procedures, and with possible side-effects. The majority of respondents were willing to join many kinds of studies, but fewer were willing to participate in first-in-human vaccine studies or studies of psychiatric drugs than in other study types. With regard to procedures, a substantial proportion were unwilling to participate in studies that involved invasive procedures, such as a lumbar puncture (45.4%) and bone marrow biopsy (42.3%), but willing to participate in studies with less invasive procedures such as a computed tomography scan of the heart (86.8%), magnetic resonance imaging (87.4%), and skin allergy testing (86.8%). Although there was some variation by gender and region, the majority were willing to participate in studies with side-effects like pain (80%) or nausea and vomiting (64%), but only a minority were willing to join if the research drug would result in their having a one in a million chance of death (34.4%), a small chance of kidney damage (16.7%), or influence how their mind works (23.2%; Figure 4). Our results suggest that healthy volunteers are willing to participate in a wide range of types of phase 1 clinical trials, and express preferences for low risk and familiar studies and study procedures, preferences which are partially affected by offers of payment.


Tunable diode laser IR spectrometer for in situ measurements of the gas phase composition and particle size distribution of Titan's atmosphere.
A new instrument, the Probe Infrared Laser Spectrometer (PIRLS), is described for in situ sensing of the gas composition and particle size distribution of Titan's atmosphere on the NASA/ESA Saturn Orbiter/Titan Probe Cassini Mission. For gas composition measurements, several narrow bandwidth (0.0001 cm(-l)) tunable lead-salt diode lasers operating near 80 K at selected, mid-IR wavelengths (3-16 microm) are directed over a pathlength defined by a small reflector extending over the edge of the probe spacecraft platform; volume mixing ratios of 10(-9) should be measurable for several species of interest. A cloud particle size spectrometer using a diode laser source at 0.78 microm shares the optical path and deployed reflector; a combination of imaging and light scattering techniques will be used to determine sizes of haze and cloud particles and their number density as a function of altitude.


A pilot trial evaluating the clinical effects of prolonged storage of red cells.
The clinical consequences of prolonged storage of red cells have not been established. In this pilot study, we evaluated whether it would be feasible to provide a continuous supply of red cells stored <8 days. In addition, we examined the potential benefits attributed to "fresh" as compared to standard red cells in 66 critically ill and cardiac surgical patients. Nine patients were issued red cells but were not transfused. From the 57 remaining patients, the number of units transfused averaged 5.5 +/- 8.43 red cell units in the experimental group compared to 3.3 +/- 3.27 red cell units in the standard group (P = 0.25). The median storage time was 4 days in the experimental group compared to 19 days in the standard group (difference of 15 days; interquartile range of 12-16 days; P < 0.001). Overall, 73% of patients received red cells with storage times that corresponded to the treatment allocation more than 90% of the time. The group receiving red cells <8 days old tended to be older on average (68 +/- 8.54 yr versus 63 +/- 15.30 yr; P = 0.13) and have more comorbid illnesses (85% versus 65%; P = 0.09). In total, 27% of patients in the experimental group died or had a life-threatening complication as compared to 13% in the standard group (P = 0.31). There were no differences in prolonged respiratory, cardiovascular, or renal support after randomization (P > 0.05). A large clinical trial comparing red cell storage times is feasible and warranted given the limited available evidence.


Socio-economic differences in the association between sickness absence and mortality: the prospective DREAM study of Danish private sector employees.
To examine duration of sickness absence as a risk marker for future mortality by socio-economic position among all private sector employees in Denmark in 1998-2004. All residents in Denmark employed in the private sector receiving sickness absence compensation in 1998 were investigated in a prospective cohort study. 236 207 persons (38.2% women, 61.8% men, age range 18-65, mean age 37.8 years) alive on 1 January 2001 were included in the study. Mortality from 1 January 2001 to 31 December 2004 was assessed using national register data. Deaths in 1999 and 2000 were excluded to determine the status of sickness absence duration as an early risk marker. For analyses within occupational grades, data were available for a sub-population of 137 607 study participants. 3040 persons died during follow-up. The age-adjusted risk of future mortality increased by duration of sickness absence in a graded fashion among men and non-blue collar workers. Among women and blue collar workers, there was no association of mortality with duration of sickness absences below 6 weeks. However, employees with > or =6 weeks of absence compared to those with 1-week absence had a substantial excess risk of death in all groups: adjusted hazard ratio 2.2 (95% CI 1.8 to 2.7) for women, 2.1 (95% CI 1.8 to 2.4) for men, 3.7 (95% CI 1.9 to 7.2) in white collar occupations, 3.3 (95% CI 2.2 to 5.0) in intermediate grade occupations and 2.0 (95% CI 1.7 to 2.3) in blue collar occupations. Administratively collected data on sickness absence compensation for periods > or =6 weeks identified "at risk" groups for future excess mortality in male and female private sector employees across occupational grade levels.


Staphylococcus aureus biofilms prevent macrophage phagocytosis and attenuate inflammation in vivo.
Biofilms are complex communities of bacteria encased in a matrix composed primarily of polysaccharides, extracellular DNA, and protein. Staphylococcus aureus can form biofilm infections, which are often debilitating due to their chronicity and recalcitrance to antibiotic therapy. Currently, the immune mechanisms elicited during biofilm growth and their impact on bacterial clearance remain to be defined. We used a mouse model of catheter-associated biofilm infection to assess the functional importance of TLR2 and TLR9 in the host immune response during biofilm formation, because ligands for both receptors are present within the biofilm. Interestingly, neither TLR2 nor TLR9 impacted bacterial density or inflammatory mediator secretion during biofilm growth in vivo, suggesting that S. aureus biofilms circumvent these traditional bacterial recognition pathways. Several potential mechanisms were identified to account for biofilm evasion of innate immunity, including significant reductions in IL-1β, TNF-α, CXCL2, and CCL2 expression during biofilm infection compared with the wound healing response elicited by sterile catheters, limited macrophage invasion into biofilms in vivo, and a skewing of the immune response away from a microbicidal phenotype as evidenced by decreases in inducible NO synthase expression concomitant with robust arginase-1 induction. Coculture studies of macrophages with S. aureus biofilms in vitro revealed that macrophages successful at biofilm invasion displayed limited phagocytosis and gene expression patterns reminiscent of alternatively activated M2 macrophages. Collectively, these findings demonstrate that S. aureus biofilms are capable of attenuating traditional host proinflammatory responses, which may explain why biofilm infections persist in an immunocompetent host.


Functional activation and effective connectivity differences in adolescent marijuana users performing a simulated gambling task.
Background. Adolescent marijuana use is associated with structural and functional differences in forebrain regions while performing memory and attention tasks. In the present study, we investigated neural processing in adolescent marijuana users experiencing rewards and losses. Fourteen adolescents with frequent marijuana use (>5 uses per week) and 14 nonuser controls performed a computer task where they were required to guess the outcome of a simulated coin flip while undergoing magnetic resonance imaging. Results. Across all participants, "Wins" and "Losses" were associated with activations including cingulate, middle frontal, superior frontal, and inferior frontal gyri and declive activations. Relative to controls, users had greater activity in the middle and inferior frontal gyri, caudate, and claustrum during "Wins" and greater activity in the anterior and posterior cingulate, middle frontal gyrus, insula, claustrum, and declive during "Losses." Effective connectivity analyses revealed similar overall network interactions among these regions for users and controls during both "Wins" and "Losses." However, users and controls had significantly different causal interactions for 10 out of 28 individual paths during the "Losses" condition. Conclusions. Collectively, these results indicate adolescent marijuana users have enhanced neural responses to simulated monetary rewards and losses and relatively subtle differences in effective connectivity.


Interventions to Attenuate Cardiovascular Calcification Progression: A Systematic Review of Randomized Clinical Trials.
Cardiovascular calcification, characterized by deposition of calcium phosphate in the arterial wall and heart valves, is associated with cardiovascular morbidity and mortality and is commonly seen in aging, diabetes, and chronic kidney disease. Whether evidence-based interventions could significantly attenuate cardiovascular calcification progression remains uncertain. We conducted a systematic review of randomized controlled trials involving interventions, compared with placebo, another comparator, or standard of care, to attenuate cardiovascular calcification. Included clinical trials involved participants without chronic kidney disease, and the outcome was cardiovascular calcification measured using radiological methods. Quality of evidence was determined by the Cochrane risk of bias and Grading of Recommendations, Assessment, Development, and Evaluations assessment. Forty-nine randomized controlled trials involving 9901 participants (median participants 104, median duration 12 months) were eligible for inclusion. Trials involving aged garlic extract (n=6 studies) consistently showed attenuation of cardiovascular calcification. Trials involving 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitors (n=14), other lipid-lowering agents (n=2), hormone replacement therapies (n=3), vitamin K (n=5), lifestyle measures (n=4), and omega-3 fatty acids (n=2) consistently showed no attenuation of cardiovascular calcification with these therapies. Trials involving antiresorptive (n=2), antihypertensive (n=2), antithrombotic (n=4), and hypoglycemic agents (n=3) showed mixed results. Singleton studies involving salsalate, folate with vitamin B6 and 12, and dalcetrapib showed no attenuation of cardiovascular calcification. Overall, Cochrane risk of bias was moderate, and the Grading of Recommendations, Assessment, Development, and Evaluations assessment for a majority of analyses was moderate certainty of evidence. Currently, there are insufficient or conflicting data for interventions evaluated in clinical trials for mitigation of cardiovascular calcification. Therapy involving aged garlic extract appears most promising, but evaluable studies were small and of short duration.


Troponins in Heart Failure - a Perpetual Challenge.
Increased troponin levels in HF are a frequent and significant finding, as it strongly correlates with the underlying pathogenic mechanisms, diagnosis and prognosis. The advent of hs-cTn testing, as opposed to conventional troponin testing, led to additional difficulties in result interpretation. Most frequently, though not exclusively, increased cTn levels in acute or chronic failure is correlated, with myocardial necrosis (AMI); the diagnosis of AMI is confirmed if other criteria are fulfilled, as described in the fourth Universal Definition of Myocardial Infarction. Increased cTn levels below the cut-off for AMI suggest acute or chronic injury, depending on the ascending and/or descending trend curve or stable levels of cTn on serial testing. In acute or chronic HF with reduced or preserved EF, increased cTn levels carry prognostic value for adverse outcomes. Acute and chronic HF, as well as other ischemic or non-ischemic conditions, may lead to a transient increase in cTn levels: hypertensive crises, tachyarrhythmias, valvular regurgitation, myocarditis, stroke, mandating differential diagnosis with ACS. There are multiple mechanisms that explain increased levels of cTn: myocardial necrosis or coronary thrombosis (type I MI), supply-demand mismatch with subendocardial ischemia/injury, cardiomyocyte apoptosis, inflammatory cytokines, neurohomonal changes. Screening for cTn levels in the population at high cardiovascular risk yields prognostic information on development of de novo HF or other cardiovascular adverse events.


Lichen Planus Pemphigoides Mimicking Toxic Epidermal Necrolysis.
Lichen planus pemphigoides (LPP) is an immunobullous disorder characterized by a combination of lesions resembling bullous pemphigoid and lichen planus (LP). Immunofluorescence studies typically show linear deposition of IgG and C3 along the basement membrane zone and circulating antibodies to the hemidesmosome protein BP180 also known as type XVII collagen. It is now recognized that drug-induced linear IgA disease may present with lesions mimicking toxic epidermal necrolysis (TEN). However, to date, there have been no reported cases of LPP presenting with TEN-like lesions. This report describes a 70-year-old woman with LPP who presented with extensive erosions mimicking TEN. This case also highlights the need to consider important etiologies such as drug-induced and paraneoplastic LPP.


The economic burden of stomas in the UK: a retrospective observational study of health records and hospital encounters.
Stomas divert waste from the small intestine (ileostomy), large intestine (colostomy) or ureters (urostomy), and complications are common. This study evaluated healthcare resource utilisation (HCRU) and costs of stomas from a UK perspective. This was a retrospective observational study of adults with new stomas (New Stoma Group) or new/existing stomas and >6 months of follow-up (Established Stoma Group) using health records linked with hospital encounters (January 2009-December 2018). Age- and sex-matched controls were identified for each stoma case (1:50). Both the New (n=8533) and Established (n=9397) stoma groups had significantly higher HCRU (all P<0.0001) and associated costs (all P<0.01), driven by inpatient admissions. New Stoma Group: colostomy versus controls, £3227 versus £99 per person; ileostomy, £2576 versus £78 per person; and urostomy, £2850 versus £110 per person (all P<0.0001). Findings were similar in the Established Stoma Group. Stomas are associated with a substantial economic burden in the UK driven by hospital care. (Supplementary data tables can be obtained from the authors.).


Descending Aortopexy and Posterior Tracheopexy for Severe Tracheomalacia and Left Mainstem Bronchomalacia.
Posterior descending aortopexy can relieve posterior intrusion of the left mainstem bronchus that may limit the effectiveness of posterior tracheobronchopexy. We review outcomes of patients undergoing both descending aortopexy and posterior tracheopexy for severe tracheobronchomalacia with posterior intrusion and left mainstem compression to determine if there were resolution of clinical symptoms and bronchoscopic evidence of improvement in airway collapse. All patients who underwent both descending aortopexy and posterior tracheopexy from October 2012 to October 2016 were retrospectively reviewed. Clinical symptoms, tracheomalacia scores based on standardized dynamic airway evaluation by anatomical region, and persistent airway intrusion requiring reoperation were collected. Data were analyzed by Wald and Wilcoxon signed-rank tests. Thirty-two patients underwent descending aortopexy and posterior tracheopexy at median age of 18 months (interquartile range 6-40 months). Median follow-up was 3 months (interquartile range 1-7 months). There were statistically significant improvements in clinical symptoms postoperatively, including cough, noisy breathing, prolonged and recurrent respiratory infections, ventilator dependence, blue spells, and brief resolved unexplained events (all P < 0.001), as well as exercise intolerance (P = 0.033), transient respiratory distress requiring positive pressure (P = 0.003), and oxygen dependence (P = 0.007). Total tracheomalacia scores improved significantly (P < 0.001), with significant segmental improvements in the middle (P = 0.003) and lower (P < 0.001) trachea, and right (P = 0.011) and left (P < 0.001) mainstem bronchi. Two patients (6%) had persistent airway intrusion requiring reoperation with anterior aortopexy or tracheopexy. Descending aortopexy and posterior tracheopexy are effective in treating severe tracheobronchomalacia and left mainstem intrusion with significant improvements in clinical symptoms and degree of airway collapse on bronchoscopy.


[Screening and follow-up for cervical cancer prevention in rural Mexico using visual inspection].
To compare the standard cervical cancer screening procedure--the Papanicolaou test or Pap smear--with detection through visual inspection using acetic acid (VIA), and visual inspection with acetic acid assisted by Aviscope (VIAM). The study was conducted between October 1998 and December 2000, in two Mixteca regions in Oaxaca, Mexico. A field trial design was used to randomize the two regions to either of the two inspection methods. In one region 2,240 women received VIA; in the other, 2,542 women received VIAM. Women positive to visual inspection and a subsample of women with negative results were referred for colposcopy and, if necessary, cervical biopsy. Statistical data analysis consisted of descriptive statistics and comparison of groups using chi-squared tests. Diagnostic values were obtained for VIA and VIAM, using as gold standards colposcopy and biopsy. VIAM identified a higher proportion of women with a cervical abnormality (16.3%) than VIA (3.4%), as well as normal women (58.5% vs. 53.8%).VIA identified a higher proportion of women with benign changes (41.2%) than VIAM (19.6%).VIAM had a greater sensitivity (p>0.05) but lower specificity (p<0.05) than VIA, using as gold standards colposcopy and biopsy. The two study groups were comparable for age, parity and cytological results; however, visual inspection results were different between the groups: VIAM identified more lesions confirmed by biopsy. The diagnostic utility of VIAM and VIA was lower than expected. Training of clinical personnel in visual inspection is critical to improve the effectiveness of these screening methods.


Responses of the competent and incompetent lower oesophageal sphincter to pentagastrin and abdominal compression.
Responses of the lower oesophageal sphincter (LOS) to pentagastrin, given by continuous intravenous infusion in doses ranging between 0 and 9 mug/kg/h, and to external abdominal compression were measured by infused catheters in healthy subjects and in a group of patients with reflux oesophagitis. In separate experiments, pressures were measured both by sensors stationary within the LOS, and by repeated continuous withdrawals of sensors from stomach to distal oesophagus. In normal subjects, doses of pentagastrin within the physiological range (0.9 mug/kg/h) produced modest but statistically significant increases in LOS pressure. By comparison, sphincteric responses in patients with oesophagitis were small and a dose of 4-5 mug/kg/h was the lowest that produced a significant increase in LOS pressure. During abdominal compression increases in LOS pressure did not significantly exceed increases in intragastric pressure in either patients or normal subjects. This was so at all doses of pentagastrin that were tested. Hence, there was no evidence of synergism between the effects of pentagastrin and abdominal compression upon the LOS. We infer from our findings that gastrin does play a modest role in the physiological regulation of human LOS tone. Relative insensitivity of the incompetent LOS to pentagastrin represents, we believe, sphincteric muscle failure. Our results are not consistent with the hypothesis that LOS incompetence is due to loss or impairment of an adaptive response of the LOS to alterations in intra-abdominal pressure.


Differential protein expression profiling in BSE disease.
Bovine spongiform encephalopathy (BSE) is a fatal neurodegenerative disease affecting cattle. Current tests for the detection of BSE are based solely on the only definitive marker of the disease, an abnormal conformer (PrP(d)), of the host encoded prion protein (PrP(c)). Recent evidence that other transmissible spongiform encephalopathy diseases can be present in the absence of PrP(d), coupled with the need to establish pre-mortem diagnostic assays have led to a search for alternative diagnostic approaches. In this study we apply differential protein expression profiling for the prediction of BSE disease in post-mortem bovine brain tissue. The protein profiles of groups of 27 BSE diseased cattle were compared with 28 control animals. Analysis using statistical learning (and linear discriminant analysis) techniques established protein markers of disease with good predictive power (sensitivity 85% and specificity 71%). Further work will be required to test the predictive markers in a wider range of diseases, particularly other neurological conditions.


Population age and household structures shape transmission dynamics of emerging infectious diseases: a longitudinal microsimulation approach.
Host population demographics and patterns of host-to-host interactions are important drivers of heterogeneity in infectious disease transmission. To improve our understanding of how population structures and changes therein influence disease transmission dynamics at the individual and population level, we model a dynamic age- and household-structured population using longitudinal microdata drawn from Belgian census and population registers. At different points in time, we simulate the spread of a close-contact infectious disease and vary the age profiles of infectiousness and susceptibility to reflect specific infections (e.g. influenza and SARS-CoV-2) using a two-level mixing model, which distinguishes between exposure to infection in the household and exposure in the community. We find that the strong relationship between age and household structures, in combination with social mixing patterns and epidemiological parameters, shape the spread of an emerging infection. Disease transmission in the adult population in particular is to a large degree explained by differential household compositions and not just household size. Moreover, we highlight how demographic processes alter population structures in an ageing population and how these in turn affect disease transmission dynamics across population groups.


Electroacupuncture suppresses morphine reward-seeking behavior: Lateral hypothalamic orexin neurons implicated.
The orexin system specifically located in the lateral hypothalamus (LH) has been shown to play a unique role in reward seeking, drug relapse, and addiction. In this study, we investigated the effects of electroacupuncture (EA) on morphine conditioned place preference (CPP), reinstatement of extinguished CPP, and the expression of orexin in the LH. A two-chamber CPP model was used to measure the rewarding properties of morphine. 2Hz EA at points BL23 and ST36 was carried out for 30min daily before morphine injection on three conditioning days. The extinguished CPP was reinstated by systemic morphine injection. Immunohistochemistry was used to detect orexin-positive nuclei in the LH. The findings indicated that the expression of morphine-induced CPP can be effectively suppressed by EA. EA suppressed orexin-positive nuclei in the LH compared with morphine-conditioned rats. A combination of EA and orexin A antagonist (10mg/kg, i.p.) produced a significant decrease in the morphine reinstatement scores. The results indicate the possibility that orexin may be involved in the acupuncture effect on drug-seeking behavior and thereby has an important role in modulating acupuncture. The synergetic use of a protocol consisting of EA and low-dose orexin A antagonist may provide a novel treatment strategy for drug reward seeking and relapse.


Targeting antigen in mature dendritic cells for simultaneous stimulation of CD4+ and CD8+ T cells.
Due to their potent immunostimulatory capacity, dendritic cells (DC) have become the centerpiece of many vaccine regimens. Immature DC (DCimm) capture, process, and present Ags to CD4(+) lymphocytes, which reciprocally activate DCimm through CD40, and the resulting mature DC (DCmat) loose phagocytic capacity, but acquire the ability to efficiently stimulate CD8(+) lymphocytes. Recombinant vaccinia viruses (rVV) provide a rapid, easy, and efficient method to introduce Ags into DC, but we observed that rVV infection of DCimm results in blockade of DC maturation in response to all activation signals, including CD40L, monocyte-conditioned medium, LPS, TNF-alpha, and poly(I:C), and failure to induce a CD8(+) response. By contrast, DCmat can be infected with rVV and induce a CD8(+) response, but, having lost phagocytic activity, fail to process the Ag via the exogenous class II pathway. To overcome these limitations, we used the CMV protein pp65 as a model Ag and designed a gene containing the lysosomal-associated membrane protein 1 targeting sequence (Sig-pp65-LAMP1) to target pp65 to the class II compartment. DCmat infected with rVV-Sig-pp65-LAMP1 induced proliferation of pp65-specific CD4(+) clones and efficiently induced a pp65-specific CD4(+) response, suggesting that after DC maturation the intracellular processing machinery for class II remains intact for at least 16 h. Moreover, infection of DCmat with rVV-Sig-pp65-LAMP1 resulted in at least equivalent presentation to CD8(+) cells as infection with rVV-pp65. These results demonstrate that despite rVV interference with DCimm maturation, a single targeting vector can deliver Ags to DCmat for the effective simultaneous stimulation of both CD4(+) and CD8(+) cells.


Diabetic nephropathy is accelerated by farnesoid X receptor deficiency and inhibited by farnesoid X receptor activation in a type 1 diabetes model.
The pathogenesis of diabetic nephropathy is complex and involves activation of multiple pathways leading to kidney damage. An important role for altered lipid metabolism via sterol regulatory element binding proteins (SREBPs) has been recently recognized in diabetic kidney disease. Our previous studies have shown that the farnesoid X receptor (FXR), a bile acid-activated nuclear hormone receptor, modulates renal SREBP-1 expression. The purpose of the present study was then to determine if FXR deficiency accelerates type 1 diabetic nephropathy in part by further stimulation of SREBPs and related pathways, and conversely, if a selective FXR agonist can prevent the development of type 1 diabetic nephropathy. Insulin deficiency and hyperglycemia were induced with streptozotocin (STZ) in C57BL/6 FXR KO mice. Progress of renal injury was compared with nephropathy-resistant wild-type C57BL/6 mice given STZ. DBA/2J mice with STZ-induced hyperglycemia were treated with the selective FXR agonist INT-747 for 12 weeks. To accelerate disease progression, all mice were placed on the Western diet after hyperglycemia development. The present study demonstrates accelerated renal injury in diabetic FXR KO mice. In contrast, treatment with the FXR agonist INT-747 improves renal injury by decreasing proteinuria, glomerulosclerosis, and tubulointerstitial fibrosis, and modulating renal lipid metabolism, macrophage infiltration, and renal expression of SREBPs, profibrotic growth factors, and oxidative stress enzymes in the diabetic DBA/2J strain. Our findings indicate a critical role for FXR in the development of diabetic nephropathy and show that FXR activation prevents nephropathy in type 1 diabetes.


Replacing dietary soybean meal with canola meal improves production and efficiency of lactating dairy cows.
Previous research suggested that crude protein (CP) from canola meal (CM) was used more efficiently than CP from solvent soybean meal (SBM) by lactating dairy cows. We tested whether dietary CP content influenced relative effectiveness of equal supplemental CP from either CM or SBM. Fifty lactating Holstein cows were blocked by parity and days in milk into 10 squares (2 squares with ruminal cannulas) in a replicated 5×5 Latin square trial. Five squares were fed: (1) low (14.5-14.8%) CP with SBM, (2) low CP with CM, (3) low CP with SBM plus CM, (4) high (16.4-16.7%) CP with SBM, and (5) high CP with CM; the other 5 squares were fed the same diets except with rumen-protected Met plus Lys (RPML) added as Mepron (Degussa Corp., Kennesaw, GA) and AminoShure-L (Balchem Corp., New Hampton, NY), which were assumed to provide 8g/d of absorbed dl-Met and 12g/d of absorbed l-Lys. Diets contained [dry matter (DM) basis] 40% corn silage, 26% alfalfa silage, 14 to 23% corn grain, 2.4% mineral-vitamin premixes, and 29 to 33% neutral detergent fiber. Periods were 3wk (total 15wk), and data from the last week of each period were analyzed using the Mixed procedures of SAS (SAS Institute Inc., Cary, NC). The only effects of RPML were increased DM intake and milk urea N (MUN) and urinary N excretion and trends for decreased milk lactose and solids-not-fat concentrations and milk-N:N intake; no significant RPML × protein source interactions were detected. Higher dietary CP increased milk fat yield and tended to increase milk yield but also elevated MUN, urine volume, urinary N excretion, ruminal concentrations of ammonia and branched-chain volatile fatty acids (VFA), lowered milk lactose concentration and milk-N:N intake, and had no effect on milk true protein yield. Feeding CM instead of SBM increased feed intake, yields of milk, energy-corrected milk, and true protein, and milk-N:N intake, tended to increase fat and lactose yields, and reduced MUN, urine volume, and urinary N excretion. At low CP, MUN was lower and intake tended to be greater on SBM plus CM versus SBM alone, but MUN and N excretion were not reduced to the same degree as on CM alone. Interactions of parity × protein source and parity × CP concentration indicated that primiparous cows were more responsive than multiparous cows to improved supply of metabolizable protein. Replacing SBM with CM reduced ruminal ammonia and branched-chain VFA concentrations, indicating lower ruminal degradation of CM protein. Replacing SBM with CM improved milk and protein yield and N-utilization in lactating cows fed both low- and high-CP diets.


[Pulmonal sarcoidosis: A rare side effect of interferon-alpha treatment for chronic hepatitis C infection].
This is a case report of a 44-year-old woman who received a 1-year-treatment with interferon-alpha for chronic hepatitis C virus infection. 3 months after cessation of the therapy she relapsed and was successfully retreated 12 months with a combination of interferon-alpha and ribavirin thereafter. During the treatment the patient developed a typical flue-like syndrome, dry cough with exertional dyspnea that was initially interpreted as a typical side effect of interferon-alpha treatment. Due to the persisting cough, further radiologic and histologic work up was done and results were significant for interstitial sarcoidosis. After interferon-treatment was stopped the pulmonary symptoms resolved completely while pathologic changes in radiographic imaging persisted. Interferon-alpha may have contributed to the development and progression of sarcoidosis by activation of cellular immunity. Although there are only few reports about pulmonary sarcoidosis associated with interferon-alpha treatment, this entity should be included in the differential diagnosis of putative side effects of interferon-alpha therapy.


Trends in Gestational Diabetes at First Live Birth by Race and Ethnicity in the US, 2011-2019.
Gestational diabetes is associated with adverse maternal and offspring outcomes. To determine whether rates of gestational diabetes among individuals at first live birth changed from 2011 to 2019 and how these rates differ by race and ethnicity in the US. Serial cross-sectional analysis using National Center for Health Statistics data for 12 610 235 individuals aged 15 to 44 years with singleton first live births from 2011 to 2019 in the US. Gestational diabetes data stratified by the following race and ethnicity groups: Hispanic/Latina (including Central and South American, Cuban, Mexican, and Puerto Rican); non-Hispanic Asian/Pacific Islander (including Asian Indian, Chinese, Filipina, Japanese, Korean, and Vietnamese); non-Hispanic Black; and non-Hispanic White. The primary outcomes were age-standardized rates of gestational diabetes (per 1000 live births) and respective mean annual percent change and rate ratios (RRs) of gestational diabetes in non-Hispanic Asian/Pacific Islander (overall and in subgroups), non-Hispanic Black, and Hispanic/Latina (overall and in subgroups) individuals relative to non-Hispanic White individuals (referent group). Among the 12 610 235 included individuals (mean [SD] age, 26.3 [5.8] years), the overall age-standardized gestational diabetes rate significantly increased from 47.6 (95% CI, 47.1-48.0) to 63.5 (95% CI, 63.1-64.0) per 1000 live births from 2011 to 2019, a mean annual percent change of 3.7% (95% CI, 2.8%-4.6%) per year. Of the 12 610 235 participants, 21% were Hispanic/Latina (2019 gestational diabetes rate, 66.6 [95% CI, 65.6-67.7]; RR, 1.15 [95% CI, 1.13-1.18]), 8% were non-Hispanic Asian/Pacific Islander (2019 gestational diabetes rate, 102.7 [95% CI, 100.7-104.7]; RR, 1.78 [95% CI, 1.74-1.82]), 14% were non-Hispanic Black (2019 gestational diabetes rate, 55.7 [95% CI, 54.5-57.0]; RR, 0.97 [95% CI, 0.94-0.99]), and 56% were non-Hispanic White (2019 gestational diabetes rate, 57.7 [95% CI, 57.2-58.3]; referent group). Gestational diabetes rates were highest in Asian Indian participants (2019 gestational diabetes rate, 129.1 [95% CI, 100.7-104.7]; RR, 2.24 [95% CI, 2.15-2.33]). Among Hispanic/Latina participants, gestational diabetes rates were highest among Puerto Rican individuals (2019 gestational diabetes rate, 75.8 [95% CI, 71.8-79.9]; RR, 1.31 [95% CI, 1.24-1.39]). Gestational diabetes rates increased among all race and ethnicity subgroups and across all age groups. Among individuals with a singleton first live birth in the US from 2011 to 2019, rates of gestational diabetes increased across all racial and ethnic subgroups. Differences in absolute gestational diabetes rates were observed across race and ethnicity subgroups.


A multifaceted approach to psychodrama in a day treatment center.
Staff at the day treatment center at Norristown (Pa.) State Hospital have found several ways of using psychodrama to facilitate information gathering, diagnostic decision-making, and treatment planning and implementation. They use behavioral techniques such as role rehearsal and modeling. In addition they use techniques such as the mirror image, in vivo dramas, and slight deception to facilitate catharsis. These approaches, which broaden the applicability of psychodrama, are illustrated by case examples.


Regulation of extracellular alkaline protease activity by histidine in a collagenolytic Vibrio alginolyticus strain.
Vibrio alginolyticus synthesized an inducible extracellular collagenase in a peptone medium during the stationary growth phase. These cultures also possessed extracellular alkaline serine protease activity. The alkaline protease activity did not require a specific inducer and it was produced in tryptone or minimal media. The collagenase was not produced in either the tryptone or minimal media. The alkaline protease activity was sensitive to catabolite repression by a number of carbon sources, including glucose, and by amino acids and ammonium ions. Cyclic AMP, dibutyryl cyclic AMP and cyclic GMP did not relieve catabolite repression. Histidine and urocanic acid stimulated the production of alkaline protease activity in tryptone and minimal media. Other compounds associated with the histidine utilization (hut) pathway did not increase alkaline protease activity. Histidine reversed the repression of alkaline protease activity by glucose of (NH4)2SO4 in minimal medium. Histidine and the compounds associated with the hut pathway inhibited collagenase production.


Multimodal Label-Free Monitoring of Adipogenic Stem Cell Differentiation Using Endogenous Optical Biomarkers.
Stem cell-based therapies carry significant promise for treating human diseases. However, clinical translation of stem cell transplants for effective treatment requires precise non-destructive evaluation of the purity of stem cells with high sensitivity (<0.001% of the number of cells). Here, a novel methodology using hyperspectral imaging (HSI) combined with spectral angle mapping-based machine learning analysis is reported to distinguish differentiating human adipose-derived stem cells (hASCs) from control stem cells. The spectral signature of adipogenesis generated by the HSI method enables identifying differentiated cells at single-cell resolution. The label-free HSI method is compared with the standard techniques such as Oil Red O staining, fluorescence microscopy, and qPCR that are routinely used to evaluate adipogenic differentiation of hASCs. HSI is successfully used to assess the abundance of adipocytes derived from transplanted cells in a transgenic mice model. Further, Raman microscopy and multiphoton-based metabolic imaging is performed to provide complementary information for the functional imaging of the hASCs. Finally, the HSI method is validated using matrix-assisted laser desorption/ionization-mass spectrometry imaging of the stem cells. The study presented here demonstrates that multimodal imaging methods enable label-free identification of stem cell differentiation with high spatial and chemical resolution.


A formal risk-benefit framework for genomic tests: facilitating the appropriate translation of genomics into clinical practice.
Evaluation of genomic tests is often challenging because of the lack of direct evidence of clinical benefit compared with usual care and unclear evidence requirements. To address these issues, this study presents a risk-benefit framework for assessing the health-related utility of genomic tests. We incorporated approaches from a variety of established fields including decision science, outcomes research, and health technology assessment to develop the framework. Additionally, we considered genomic test stakeholder perspectives and case studies. We developed a three-tiered framework: first, we use decision-analytic modeling techniques to synthesize data, project incidence of clinical events, and assess uncertainty. Second, we defined the health-related utility of genomic tests as improvement in health outcomes as measured by clinical event rates, life expectancy, and quality-adjusted life-years. Finally, we displayed results using a risk-benefit policy matrix to facilitate the interpretation and implementation of findings from these analyses. A formal risk-benefit framework may accelerate the utilization and practice-based evidence development of genomic tests that pose low risk and offer plausible clinical benefit, while discouraging premature use of tests that provide little benefit or pose significant health risks compared with usual care.


Ketogenic Diet for KARS-Related Mitochondrial Dysfunction and Progressive Leukodystrophy.
KARS encodes lysyl-tRNA synthetase, which is essential for protein translation. KARS mutations sometimes cause impairment of cytoplasmic and mitochondrial protein synthesis, and sometimes lead to progressive leukodystrophies with mitochondrial signature and psychomotor regression, and follow a rapid regressive course to premature death. There has been no disease-modifying therapy beyond supportive treatment. We present a 5-year-old male patient with an asymmetrical leukodystrophy who showed overt evidence of mitochondrial dysfunction, including elevation of lactate on brain MR spectroscopy and low oxygen consumption rate in fibroblasts. We diagnosed this patient's condition as KARS-related leukodystrophy with cerebral calcification, congenital deafness, and evidence of mitochondrial dysfunction. We employed a ketogenic diet as well as multiple vitamin supplementation with the intention to alleviate mitochondrial dysfunction. The patient showed alleviation of his psychomotor regression and even partial restoration of his abilities within 4 months. This is an early report of a potential disease-modifying therapy for KARS-related progressive leukodystrophy without appreciable adverse effects.


Quantitative Chemotherapeutic Profiling of Gynecologic Cancer Cell Lines Using Approved Drugs and Bioactive Compounds.
Heterogeneous response to chemotherapy is a major issue for the treatment of cancer. For most gynecologic cancers including ovarian, cervical, and placental, the list of available small molecule therapies is relatively small compared to options for other cancers. While overall cancer mortality rates have decreased in the United States as early diagnoses and cancer therapies have become more effective, ovarian cancer still has low survival rates due to the lack of effective treatment options, drug resistance, and late diagnosis. To understand chemotherapeutic diversity in gynecologic cancers, we have screened 7914 approved drugs and bioactive compounds in 11 gynecologic cancer cell lines to profile their chemotherapeutic sensitivity. We identified two HDAC inhibitors, mocetinostat and entinostat, as pan-gynecologic cancer suppressors with IC50 values within an order of magnitude of their human plasma concentrations. In addition, many active compounds identified, including the non-anticancer drugs and other compounds, diversely inhibited the growth of three gynecologic cancer cell groups and individual cancer cell lines. These newly identified compounds are valuable for further studies of new therapeutics development, synergistic drug combinations, and new target identification for gynecologic cancers. The results also provide a rationale for the personalized chemotherapeutic testing of anticancer drugs in treatment of gynecologic cancer.


Exosome derived from murine adipose-derived stromal cells: Neuroprotective effect on in vitro model of amyotrophic lateral sclerosis.
Therapeutic strategies for the fatal neurodegenerative disease amyotrophic lateral sclerosis (ALS) have not yet provided satisfactory results. Interest in stem cells for the treatment of neurodegenerative diseases is increasing and their beneficial action seems to be due to a paracrine effect via the release of exosomes, main mediators of cell-cell communication. Here we wished to assess, in vitro, the efficacy of a novel non-cell therapeutic approach based on the use of exosomes derived from murine adipose-derived stromal cells on motoneuron-like NSC-34 cells expressing ALS mutations, and used as in vitro models of disease. In particular, we set out to investigate the effect of exosomes on NSC-34 naïve cells and NSC-34 cells overexpressing human SOD1(G93A) or SOD1(G37R) or SOD1(A4V) mutants, exposed to oxidative stress. The data presented here indicate for the first time that exosomes (0.2 µg/ml) are able to protect NSC-34 cells from oxidative damage, which is one of the main mechanism of damage in ALS, increasing cell viability. These data highlight a promising role of exosomes derived from stem cells for potential therapeutic applications in motoneuron disease.


Introduction of non-invasive prenatal testing as a first-tier aneuploidy screening test: A survey among Dutch midwives about their role as counsellors.
In 2014, non-invasive prenatal testing (NIPT) for trisomies 21, 18 and 13 was added to the Dutch prenatal screening program as part of the TRIDENT study. Most (85%) pregnant Dutch women are counselled for prenatal aneuploidy screening by primary care midwives. This will remain when NIPT is implemented as a first-tier screening test. We therefore investigated midwife counsellors': 1) Knowledge about NIPT; 2) Attitudes towards NIPT as first-tier screening test; and 3) Experiences with informing clients about NIPT. Between April-June 2015, an online questionnaire to assess knowledge about NIPT, attitudes towards NIPT, and experiences with NIPT was completed by 436 Dutch primary care midwives. We found that 59% midwives answered ≥7 of 8 knowledge questions correctly. Continuing professional education attendance and more positive attitudes towards prenatal screening for Down syndrome were positively associated with the total knowledge score (β = 0.261; p = 0.007 and β = 0.204; p = 0.015, respectively). The majority (67%) were in favor of replacing First trimester Combined Test with NIPT, although 41% preferred to maintain a nuchal translucency measurement alongside NIPT. We conclude that midwives demonstrated solid knowledge about NIPT that may still be improved in some areas. Dutch midwives overwhelmingly support the integration of NIPT as a first-tier screening test.


Inhibition of Na(+)-independent H+ pump by Na(+)-induced changes in cell Ca2+.
Apical membrane H+ extrusion in the renal outer medullary collecting duct, inner stripe, is mediated by a Na(+)-independent H+ pump. To examine the regulation of this transporter, cell pH and cell Ca2+ were measured microfluorometrically in in vitro perfused tubules using 2',7'-bis(carboxyethyl)-5(6)-carboxyfluorescein and fura-2, respectively. Apical membrane H+ pump activity, assayed as cell pH recovery from a series of acid loads (NH3/NH+4 prepulse) in the total absence of ambient Na+, initially occurred at a slow rate (0.06 +/- 0.02 pH units/min), which was not sufficient to account for physiologic rates of H+ extrusion. Over 15-20 min after the initial acid load, the rate of Na(+)-independent cell pH recovery increased to 0.63 +/- 0.09 pH units/min, associated with a steady-state cell pH greater than the initial pre-acid load cell pH. This pattern suggested an initial suppression followed by a delayed activation of the apical membrane H+ pump. Replacement of peritubular Na+ with choline or N-methyl-D-glucosamine resulted in an initial spike increase in cell Ca2+ followed by a sustained increase in cell Ca2+. The initial rate of Na(+)-independent cell pH recovery could be increased by elimination of the Na+ removal-induced sustained cell Ca2+ elevation by: (a) performing studies in the presence of 135 mM peritubular Na+ (1 mM peritubular amiloride used to inhibit basolateral membrane Na+/H+ antiport); (b) clamping cell Ca2+ low with dimethyl-BAPTA, an intracellular Ca2+ chelating agent; or (c) removal of extracellular Ca2+. Cell acidification induced a spike increase in cell Ca2+. The late acceleration of Na(+)-independent cell pH recovery was independent of Na+ removal and of the method used to acidify the cell, but was eliminated by prevention of the cell Ca2+ spike and markedly delayed by the microfilament-disrupting agent, cytochalasin B. This study demonstrates that peritubular Na+ removal results in a sustained elevation in cell Ca2+, which inhibits the apical membrane H+ pump. In addition, rapid cell acidification associated with a spike increase in cell Ca2+ leads to a delayed activation of the H+ pump. Thus, cell Ca2+ per se, or a Ca(2+)-activated pathway, can modulate H+ pump activity.


Predictive coding of dynamical variables in balanced spiking networks.
Two observations about the cortex have puzzled neuroscientists for a long time. First, neural responses are highly variable. Second, the level of excitation and inhibition received by each neuron is tightly balanced at all times. Here, we demonstrate that both properties are necessary consequences of neural networks that represent information efficiently in their spikes. We illustrate this insight with spiking networks that represent dynamical variables. Our approach is based on two assumptions: We assume that information about dynamical variables can be read out linearly from neural spike trains, and we assume that neurons only fire a spike if that improves the representation of the dynamical variables. Based on these assumptions, we derive a network of leaky integrate-and-fire neurons that is able to implement arbitrary linear dynamical systems. We show that the membrane voltage of the neurons is equivalent to a prediction error about a common population-level signal. Among other things, our approach allows us to construct an integrator network of spiking neurons that is robust against many perturbations. Most importantly, neural variability in our networks cannot be equated to noise. Despite exhibiting the same single unit properties as widely used population code models (e.g. tuning curves, Poisson distributed spike trains), balanced networks are orders of magnitudes more reliable. Our approach suggests that spikes do matter when considering how the brain computes, and that the reliability of cortical representations could have been strongly underestimated.


Conceptual model for physical therapist management of chronic fatigue syndrome/myalgic encephalomyelitis.
Fatigue is one of the most common reasons why people consult health care providers. Chronic fatigue syndrome/myalgic encephalomyelitis (CFS/ME) is one cause of clinically debilitating fatigue. The underdiagnosis of CFS/ME, along with the spectrum of symptoms that represent multiple reasons for entry into physical therapy settings, places physical therapists in a unique position to identify this health condition and direct its appropriate management. The diagnosis and clinical correlates of CFS/ME are becoming better understood, although the optimal clinical management of this condition remains controversial. The 4 aims of this perspective article are: (1) to summarize the diagnosis of CFS/ME with the goal of promoting the optimal recognition of this condition by physical therapists; (2) to discuss aerobic system and cognitive deficits that may lead to the clinical presentation of CFS/ME; (3) to review the evidence for graded exercise with the goal of addressing limitations in body structures and functions, activity, and participation in people with CFS/ME; and (4) to present a conceptual model for the clinical management of CFS/ME by physical therapists.


Influence of knowledge and beliefs on consumption of performance enhancing agents in north-western Saudi Arabia.
Consumption of performance enhancing agents (PEAs) has a wide range of negative health consequences, but knowledge of these consequences among gym users of PEAs in Saudi Arabia is not well understood. Identify the knowledge, awareness, beliefs and attitudes of gym users about negative health consequences of using PEAs, and the relationship between these factors and use of these agents. Cross-sectional study. Five gyms in Madinah city, Saudi Arabia. Convenience sampling was used to recruit gym users. An electronic self-administered questionnaire was used to collect data. Level of knowledge about the negative health consequences of PEAs among gym users. About 70% of 316 participants had used one or more of PEAs over the last six months. Of those, about 68.4% used protein powder supplements and 48.1% used energy drinks. Participants who believed that protein powder supplements (c2=52.3, P < .01) and energy drinks (c2=35.2, P < .01) had health hazards used these agents less often than others during the six months preceding data collection. Participants who had less knowledge about the negative health consequences were more likely to use protein powder supplement (t=2.38, P=.018). On the other hand, those who were more knowledgeable about the negative health consequences of insulin, were more likely to use insulin (t=2.45, P=.015). Misuse of PEAs is widespread among gym users in Saudi Arabia. Improving the level of knowledge and awareness of possible serious health consequences would hopefully lead to reduced PEA consumption. The temporal sequence of cause and effect could not be determined in a cross sectional study. Convenience sampling in a single city limited the generalizability of the findings to all regions of Saudi Arabia.


Structural Mechanism for GSDMD Targeting by Autoprocessed Caspases in Pyroptosis.
The pyroptosis execution protein GSDMD is cleaved by inflammasome-activated caspase-1 and LPS-activated caspase-11/4/5. The cleavage unmasks the pore-forming domain from GSDMD-C-terminal domain. How the caspases recognize GSDMD and its connection with caspase activation are unknown. Here, we show site-specific caspase-4/11 autoprocessing, generating a p10 product, is required and sufficient for cleaving GSDMD and inducing pyroptosis. The p10-form autoprocessed caspase-4/11 binds the GSDMD-C domain with a high affinity. Structural comparison of autoprocessed and unprocessed capase-11 identifies a β sheet induced by the autoprocessing. In caspase-4/11-GSDMD-C complex crystal structures, the β sheet organizes a hydrophobic GSDMD-binding interface that is only possible for p10-form caspase-4/11. The binding promotes dimerization-mediated caspase activation, rendering a cleavage independently of the cleavage-site tetrapeptide sequence. Crystal structure of caspase-1-GSDMD-C complex shows a similar GSDMD-recognition mode. Our study reveals an unprecedented substrate-targeting mechanism for caspases. The hydrophobic interface suggests an additional space for developing inhibitors specific for pyroptotic caspases.


Oesophageal motor changes in diabetes mellitus.
Radiography and manometry have been used to study oesophageal motor function in a group of 31 diabetics, 23 of whom had alimentary or genitourinary symptoms attributed to autonomic neuropathy. Peristalsis was of diminished amplitude and oesophageal emptying in the 15 degrees Trendelen-Peristalsis was of diminished amplitude and oesophageal emptying in the 15 degrees Trendelenburg position was delayed. The lower oesophageal sphincter pressure was reduced. The changes seldom caused symptoms and they were not confined to those diabetics with alimentary or genitourinary symptoms attributable to autonomic neuropathy. They suggest that autonomic neuropathy in diabetes is widespread and often subclinical. Degeneration of the ganglion cells of the oesophageal myenteric plexus is associated with hypersensitivity of the oesophageal smooth muscle to cholinergic agents. Bethanecol, a cholinergic drug with muscarinic actions, accelerated oesophageal emptying and increased the lower oesophageal sphincter pressure to normal levels in our diabetics but the hypersensitivity to this drug found in the presence of ganglion cell degeneration was not seen. This implies that in diabetic autonomic neuropathy the predominant lesion is in the preganglionic fibres of the vagus rather than in the myenteric plexus of the oesophageal wall.


Automating Model Comparison in Factor Graphs.
Bayesian state and parameter estimation are automated effectively in a variety of probabilistic programming languages. The process of model comparison on the other hand, which still requires error-prone and time-consuming manual derivations, is often overlooked despite its importance. This paper efficiently automates Bayesian model averaging, selection, and combination by message passing on a Forney-style factor graph with a custom mixture node. Parameter and state inference, and model comparison can then be executed simultaneously using message passing with scale factors. This approach shortens the model design cycle and allows for the straightforward extension to hierarchical and temporal model priors to accommodate for modeling complicated time-varying processes.


Excess phosphoserine-129 α-synuclein induces synaptic vesicle trafficking and declustering defects at a vertebrate synapse.
α-Synuclein is a presynaptic protein that regulates synaptic vesicle (SV) trafficking. In Parkinson's disease (PD) and dementia with Lewy bodies (DLB), α-synuclein aberrantly accumulates throughout neurons, including at synapses. During neuronal activity, α-synuclein is reversibly phosphorylated at serine 129 (pS129). While pS129 comprises ∼4% of total α-synuclein under physiological conditions, it dramatically increases in PD and DLB brains. The impacts of excess pS129 on synaptic function are currently unknown. We show here that compared with wild-type (WT) α-synuclein, pS129 exhibits increased binding and oligomerization on synaptic membranes and enhanced vesicle "microclustering" in vitro. Moreover, when acutely injected into lamprey reticulospinal axons, excess pS129 α-synuclein robustly localized to synapses and disrupted SV trafficking in an activity-dependent manner, as assessed by ultrastructural analysis. Specifically, pS129 caused a declustering and dispersion of SVs away from the synaptic vicinity, leading to a significant loss of total synaptic membrane. Live imaging further revealed altered SV cycling, as well as microclusters of recently endocytosed SVs moving away from synapses. Thus, excess pS129 caused an activity-dependent inhibition of SV trafficking via altered vesicle clustering/reclustering. This work suggests that accumulation of pS129 at synapses in diseases like PD and DLB could have profound effects on SV dynamics.


Cloning and expression of ornithine decarboxylase gene from human colorectal carcinoma.
To construct and express ODC recombinant gene for further exploring its potential use in early diagnosis of colorectal carcinoma. Total RNA was extracted from colon cancer tissues and amplified by reverse-transcription PCR with two primers, which span the whole coding region of ODC. The synthesized ODC cDNA was cloned into vector pQE-30 at restriction sites BamH I and Sal I which constituted recombinant expression plasmid pQE30-ODC. The sequence of inserted fragment was confirmed by DNA sequencing, the fusion protein including 6His-tag was facilitated for purification by Ni-NTA chromatographic column. ODC expression vector was constructed and confirmed with restriction enzyme digestion and subsequent DNA sequencing. The DNA sequence matching on NCBI Blast showed 99 % affinity. The vector was transformed into E. coli M15 and expressed. The expressed ODC protein was verified with Western blotting. The ODC prokaryote expression vector is constructed and thus greatly facilitates to study the role of ODC in colorectal carcinoma.


Effect of telmisartan on nitric oxide--asymmetrical dimethylarginine system: role of angiotensin II type 1 receptor gamma and peroxisome proliferator activated receptor gamma signaling during endothelial aging.
Telmisartan, in addition to blocking angiotensin (Ang) II type 1 receptor (AT(1)R), activates peroxisome proliferator activated receptor gamma (PPARgamma) signaling that interferes with nitric oxide (NO) system. Because aging of endothelial cells (ECs) is hallmarked by a reduction in NO synthesis, we hypothesized that telmisartan increases NO formation by regulated asymmetrical dimethylarginine (ADMA)-dimethylarginine dimethylaminohydrolase (DDAH)-system through blocking AT(1)R and activating PPARgamma signaling. To test this hypothesis, ECs were cultured with telmisartan, eprosartan, Ang II, and GW9662 (PPARgamma antagonist) until the twelfth passage. During the process of aging, PPARgamma protein expression decreased significantly, whereas the expression of AT(1)R increased. Telmisartan reversed these effects and dose-dependently decreased reactive oxygen species and 8-iso-prostaglandin (PG) F(2alpha) formation. This effect was associated with an upregulated activity and protein expression of DDAH, accompanied by a decrease in ADMA concentration, an increase in NO metabolites, and delayed senescence. Blockade of PPARgamma signaling by GW9662 or PPARgamma small-interference RNA prevented the effect of telmisartan on ADMA-DDAH-NO system. Coincubation with Ang II did not affect the effect of telmisartan-delayed senescence, whereas Ang II itself accelerated endothelial aging. Moreover, AT(1)R blocker eprosartan that did not influence PPARgamma protein expression had no effect on ADMA system and senescence. We have demonstrated that telmisartan mainly by activating PPARgamma signaling can alter the catabolism and release of ADMA as an important cardiovascular risk factor. We therefore propose that telmisartan translationally and posttranslationally upregulated DDAH expression via activation of PPARgamma signaling, causing ADMA to diminish and increase NO synthesis sufficient to delay senescence.


Increasing global mortality from idiopathic pulmonary fibrosis in the twenty-first century.
Evidence from the United Kingdom suggests that the number of deaths from idiopathic pulmonary fibrosis is increasing, although comparable international data are limited. We aimed to collate death certification data from multiple countries to determine global trends in mortality from idiopathic pulmonary fibrosis. Data were obtained from the national statistics agencies of countries with relevant mortality records. Age-standardized mortality rates were calculated, and Poisson regression modeling was used to calculate rate ratios. Meta-analysis was used to calculate an overall estimate of mortality change over time. Ten countries provided mortality data on pulmonary fibrosis over a period from 1999 to 2012. Age-standardized mortality ranged between 4 and 10 per 100,000 population for the most recent years of data, being lowest in Sweden (4.68 per 100,000), Spain (5.38 per 100,000), and New Zealand (5.55 per 100,000), and highest in the United Kingdom (9.84 per 100,000 in England and Wales, 10.71 per 100,000 in Scotland) and Japan (10.26 per 100,000). Positive associations with male sex and increasing age were consistently observed across all countries. There was an overall 2-3% annual increase in mortality depending on codes used for classification: For broad codes the overall rate ratio was 1.03 (95% confidence interval, 1.02-1.04; P < 0.001) and for narrow codes the overall rate ratio was 1.02 (95% confidence interval, 1.01-1.03; P < 0.001). Validation in a local cohort showed that idiopathic pulmonary fibrosis was recorded as the underlying cause of death in two-thirds of known cases and anywhere on the death certificate in 80% of cases. Mortality from idiopathic pulmonary fibrosis is increasing steadily worldwide, despite the fact that death certification will almost certainly underestimate true mortality. We estimate that there will be between 28,000 and 65,000 deaths in Europe and between 13,000 and 17,000 deaths in the United States from idiopathic pulmonary fibrosis clinical syndrome in 2014. Variation between countries remains but is less than previously reported.


Biological substrates of emotional reactivity and regulation in adolescence during an emotional go-nogo task.
Adolescence is a transition period from childhood to adulthood that is often characterized by emotional instability. This period is also a time of increased incidence of anxiety and depression, underscoring the importance of understanding biological substrates of behavioral and emotion regulation during adolescence. Developmental changes in the brain in concert with individual predispositions for anxiety might underlie the increased risk for poor outcomes reported during adolescence. We tested the hypothesis that difficulties in regulating behavior in emotional contexts in adolescents might be due to competition between heightened activity in subcortical emotional processing systems and immature top-down prefrontal systems. Individual differences in emotional reactivity might put some teens at greater risk during this sensitive transition in development. We examined the association between emotion regulation and frontoamygdala circuitry in 60 children, adolescents, and adults with an emotional go-nogo paradigm. We went beyond examining the magnitude of neural activity and focused on neural adaptation within this circuitry across time with functional magnetic resonance imaging. Adolescents showed exaggerated amygdala activity relative to children and adults. This age-related difference decreased with repeated exposures to the stimuli, and individual differences in self-ratings of anxiety predicted the extent of adaptation or habituation in amygdala. Individuals with higher trait anxiety showed less habituation over repeated exposures. This failure to habituate was associated with less functional connectivity between ventral prefrontal cortex and amygdala. These findings suggest that exaggerated emotional reactivity during adolescence might increase the need for top-down control and put individuals with less control at greater risk for poor outcomes.


Oligodeoxyribonucleotide ligation to single-stranded cDNAs: a new tool for cloning 5' ends of mRNAs and for constructing cDNA libraries by in vitro amplification.
Cloning full length cDNAs is a difficult task especially if mRNAs are not abundant or if tissue is only available in limited amounts. Current strategies are based on in vitro amplification of cDNAs after adding a homopolymeric tail at the 3' end of the ss-cDNA. Since subsequent amplification steps yield unspecific amplified DNA mostly due to non-specific annealing of the reverse primer containing a homopolymeric tail, we have devised a new strategy based on the ligation of single-stranded oligodeoxyribonucleotide to the 3' end of single-stranded cDNAs. The efficiency of the strategy was assessed by analyzing the 5' ends of the rat pineal gland tryptophan hydroxylase messenger. The 5' end of the least abundant messenger (0.005% of total mRNAs) could be cloned without selection. Sixty percent of the analyzed clones correspond to TPH. This technique revealed a 5-nt stretch not apparent using dG tailing strategy. The potentiality of the method for generating cDNAs libraries was tested with 10(4) PC12 cells. In this library, the abundance of tyrosine hydroxylase clones (0.03%) correlated well with the abundance of the corresponding messenger, showing that no major distortion was introduced into the construction of the library.


Image segmentation and registration algorithm to collect homologous landmarks for age-related thoracic morphometric analysis - biomed 2011.
Adults 65 years of age and older currently constitute more than 12% of the total population and the elderly population is projected to reach nearly 20% by 2030. Previous studies have shown that skeletal and physiological resilience decline with age, resulting in a decreased ability for the body to withstand traumatic insults. In the current study, an algorithm was developed to quantify age and gender-specific variations in the thoracic skeletal morphology. Normal chest CT scans of males and females ages 0-100 were collected from a radiological database. Image segmentation and subsequent image registration was used to collect landmark data from the ribs. Rigid, affine, and non-rigid, non-linear transformations are used to morph segmented ribs from different subjects to a "rib atlas". The atlas consists of a normal chest CT scan from an average male with over 100 landmarks placed per rib. The transformation matrices are used to map landmarks from the atlas coordinate system to the coordinate system of each CT scan, effectively allowing for collection of homologous (or comparable) rib landmarks across all subjects. Geometric morphometrics will be used in future work to analyze the landmark data to formulate age and gender-specific shape and size variation functions. These functions will be used to create a scalable finite element model of the thorax that will be used to predict thoracic injury response for different ages and genders.


Primary amyloidoma of the spine. A case report and review of the literature.
Primary amyloidosis of bone is a rare disease process. When amyloid lesions do occur in bone, they are often associated with myeloma or other plasma cell dyscrasias. Because of its rarity and its protean clinical and radiographic picture adequate biopsy tissue for histopathologic studies is essential for accurate diagnosis. This case presents a 43-year-old man with nondescript back pain and an impressive radiographic lesion in his lower thoracic spine demonstrating the need for careful diagnostic evaluation and a high incidence of suspicion. When occurring primarily in a localized anatomic area, the prognosis for survival is excellent, and cure can be expected by local excision and reconstruction, as necessary.


Paraneoplastic motor neuron disease associated with Purkinje cell autoantibody type 1.
Motor neuron disease has been reported as a rare result of systemic cancers, likely related to an autoimmune effecter mechanism. These patients have been described as having a more rapid onset of symptoms than typical motor neuron disease. Few of these reports demonstrate an association of a cancer-related autoantibody and with motor neuron disease. We present a 54-year-old woman with infiltrating ductal carcinoma of the breast who later developed lower motor neuron disease and was found to have Purkinje cell autoantibodies type 1. The association of lower motor neuron disease as a consequence of breast cancer with this autoantibody profile has not been described previously. This report emphasizes the importance of considering a paraneoplastic syndrome in a patient with an uncommon presentation of motor neuron disease.


Recombination in the B chromosome of maize to produce A-B-A chromosomes.
The translocations between the supernumerary B chromosomes and the normal A chromosomes of maize provide a valuable tool for gene localizations, dosage studies and characterization of mutants as null, leaky or gain-of-function. A procedure is described, that relies on recombination in the B chromosome, for marking each of the various B-A translocations with a single dominant marker that will allow dosage classifications of individuals at the mature kernel stage. This marker is R-scm3, which conditions anthocyanin pigment in the aleurone of the endosperm and the scutellum of the embryo. A test for recombination in the B chromosome was conducted by crossing together two translocations, that were broken on opposite sides of the B centromere, and in different A chromosome arms, namely TB-1La and TB-10L18. An example was recovered that linked genetic markers on 1L and 10L to the B centromere. Cytological examination at pachytene of meiosis confirmed the new chromosomal linkage. The use of this procedure to produce a comprehensive set of uniformly marked B-A translocations is discussed.


Infections on the move: how transient phases of host movement influence disease spread.
Animal movement impacts the spread of human and wildlife diseases, and there is significant interest in understanding the role of migrations, biological invasions and other wildlife movements in spatial infection dynamics. However, the influence of processes acting on infections during transient phases of host movement is poorly understood. We propose a conceptual framework that explicitly considers infection dynamics during transient phases of host movement to better predict infection spread through spatial host networks. Accounting for host transient movement captures key processes that occur while hosts move between locations, which together determine the rate at which hosts spread infections through networks. We review theoretical and empirical studies of host movement and infection spread, highlighting the multiple factors that impact the infection status of hosts. We then outline characteristics of hosts, parasites and the environment that influence these dynamics. Recent technological advances provide disease ecologists unprecedented ability to track the fine-scale movement of organisms. These, in conjunction with experimental testing of the factors driving infection dynamics during host movement, can inform models of infection spread based on constituent biological processes.


Enhanced contact hypersensitivity and antiviral immune responses in vivo by keratinocyte-targeted overexpression of IL-15.
IL-15 is involved in lymphocyte homeostasis. To investigate the role of IL-15 in the skin in vivo, mice were generated that overexpress IL-15 in keratinocytes, resulting in increased IL-15 protein levels in the skin but not elevated IL-15 serum concentrations. Keratin 14 (K14)-IL-15 transgenic (tg) mice showed increased contact hypersensitivity (CHS) responses. Transfer of primed wild-type (wt) and tg T cells into naive wt or tg recipients indicated that skin-derived IL-15 enhanced the induction but not the elicitation phase of CHS. Tg mice could be sensitized even by suboptimal hapten concentrations. Accordingly, Langerhans cells (LC) from tg skin were identified as potent allostimulators, suggesting the involvement of IL-15-stimulated LC in the induction of adaptive immunity. Overexpression of IL-15 also strengthened innate immunity since tg mice infected with human HSV type I developed significantly smaller HSV skin lesions. In addition, tg mice resisted re-infection with HSV more effectively than wt mice did, which was associated with an elevated anti-HSV Ab production. Accordingly, injection of serum from re-infected tg mice protected naive recipients significantly from epicutaneous HSV infection, indicating that anti-HSV Ab produced by tg mice play an important role in resistance in vivo. Together, our results show that overexpression of IL-15 in the epidermis enhances both innate and adaptive cutaneous immunity.


One-year developmental stability and covariance among oddball, novelty, go/no-go, and flanker event-related potentials in adolescence: A monozygotic twin study.
ERP measures may index genetic risk for psychopathology before disorder onset in adolescence, but little is known about their developmental rank-order stability during this period of significant brain maturation. We studied ERP stability in 48 pairs of identical twins (age 14-16 years) tested 1 year apart. Trial-averaged voltage waveforms were extracted from electroencephalographic recordings from oddball/novelty, go/no-go, and flanker tasks, and 16 amplitude measures were examined. Members of twin pairs were highly similar, whether based on ERP amplitude measures (intraclass correlation [ICC] median = .64, range = .44-.86) or three factor scores (all ICCs ≥ .69) derived from them. Stability was high overall, with 69% of the 16 individual measures generating stability coefficients exceeding .70 and all factor scores showing stability above .75. Measures from 10 difference waveforms calculated from paired conditions within tasks were also examined, and were associated with lower twin similarity (ICC median = .52, .38-.64) and developmental stability (only 30% exceeding .70). In a supplemental analysis, we found significant developmental stability for error-related negativity (range = .45-.55) and positivity (.56-.70) measures when average waveforms were based on one or more trials, and that these values were equivalent to those derived from averages using the current field recommendation, which requires six or more trials. Overall, we conclude that the studied brain measures are largely stable over 1 year of mid- to late adolescence, likely reflecting familial etiologic influences on brain functions pertaining to cognitive control and salience recognition.