New parameter in diagnosis of acute appendicitis: platelet distribution width.
To investigate the diagnostic accuracy of the mean platelet volume and platelet distribution width in acute appendicitis. This retrospective, case-controlled study compared 295 patients with acute appendicitis (Group I, 100 patients with other intra-abdominal infections (Group II), and 100 healthy individuals (Group III) between January 2012 and January 2013. The age, gender, and white blood cell count, neutrophil percentage, mean platelet volume, and platelet distribution width values from blood samples were compared among the groups. Statistical analyses were performed using SPSS for Windows 21.0 software. In addition, the sensitivity, specificity, positive and negative predictive values and likelihood ratios, and diagnostic accuracy were calculated. The mean ages of patients were 29.9 ± 12.0 years for Group I, 31.5 ± 14.0 years for Group II, and 30.4 ± 13.0 years for Group III. Demographic features such as age and gender were not significantly different among the groups. White blood cell count, neutrophil percentage and platelet distribution width were significantly higher in Group I compared to groups II and III (P < 0.05). Diagnostically, the sensitivity, specificity and diagnostic accuracy were 73.1%, 94.0%, and 78% for white blood cell count, 70.0%, 96.0%, and 76.0% for neutrophil percentage, 29.5%, 49.0%, and 34.0% for mean platelet volume, and 97.1%, 93.0%, and 96.0% for platelet distribution width, respectively. The highest diagnostic accuracy detected was for platelet distribution width between Group I and Group III (P < 0.01). Platelet distribution width analysis can be used for diagnosis of acute appendicitis without requiring additional tests, thus reducing the cost and loss of time.


Major intercontinentally distributed sequence types of Kingella kingae and development of a rapid molecular typing tool.
Although Kingella kingae is the most common etiology of osteoarticular infections in young children, is a frequent cause of bacteremia in those younger than 4 years, and has been involved in clusters of invasive infections among daycare center attendees, the population structure of the species has not been systematically studied. Using multilocus sequence typing, we investigated the genetic diversity of the largest intercontinental collection of K. kingae strains to date. To facilitate typing of bacterial isolates, we developed a novel genotyping tool that targets the DNA uptake sequence (DUS). Among 324 strains isolated from asymptomatic carriers and patients from Israel, Europe, North America, and Australia with various invasive forms of the disease from 1960 to 2013, we identified 64 sequence types (STs) and 12 ST complexes (STcs). Five predominant STcs, comprising 72.2% of all strains, were distributed intercontinentally. ST-6 was the most frequent, showing a worldwide distribution, and appeared genotypically isolated by exhibiting few neighboring STs, suggesting an optimal fitness. ST-14 and ST-23 appeared to be the oldest groups of bacteria, while ST-25 probably emerged more recently from the highly evolutive ST-23. Using the DUS typing method, randomly chosen isolates were correctly classified to one of the major STcs. The comprehensive description of K. kingae evolution would help to detect new emerging clones and decipher virulence and fitness mechanisms. The rapid and reproducible DUS typing method may serve in the initial investigation of K. kingae outbreaks.


Pulmonary artery enlargement is associated with right ventricular dysfunction and loss of blood volume in small pulmonary vessels in chronic obstructive pulmonary disease.
Chronic obstructive pulmonary disease causes significant morbidity and concomitant pulmonary vascular disease and cardiac dysfunction are associated with poor prognosis. Computed tomography-detected relative pulmonary artery (PA) enlargement defined as a PA to ascending aorta diameter ratio >1 (PA:A>1) is a marker for pulmonary hypertension and predicts chronic obstructive pulmonary disease exacerbations. However, little is known about the relationship between the PA:A ratio, pulmonary blood volume, and cardiac function. A single-center prospective cohort study of patients with chronic obstructive pulmonary disease was conducted. Clinical characteristics and computed tomography metrics, including the PA:A and pulmonary blood vessel volume, were measured. Ventricular functions, volumes, and dimensions were measured by cine cardiac MRI with 3-dimensional analysis. Linear regression examined the relationships between clinical characteristics, computed tomography and cardiac MRI metrics, and 6-minute walk distance. Twenty-four patients were evaluated and those with PA:A>1 had higher right ventricular (RV) end-diastolic and end-systolic volume indices accompanied by lower RV ejection fraction (52±7% versus 60±9%; P=0.04). The PA:A correlated inversely with total intraparenchymal pulmonary blood vessel volume and the volume of distal vessels with a cross-sectional area of <5 mm(2). Lower forced expiratory volume, PA:A>1, and hyperinflation correlated with reduced RV ejection fraction. Both PA diameter and reduced RV ejection fraction were independently associated with reduced 6-minute walk distance. The loss of blood volume in distal pulmonary vessels is associated with PA enlargement on computed tomography. Cardiac MRI detects early RV dysfunction and remodeling in nonsevere chronic obstructive pulmonary disease patients with a PA:A>1. Both RV dysfunction and PA enlargement are independently associated with reduced walk distance. URL: http://www.clinicaltrials.gov. Unique identifier: NCT00608764.


Effects of topiramate on cognitive function.
To explore the impact of topiramate on tests of intellect and other cognitive processes. This was a retrospective study. The neuropsychological test scores of 18 patients obtained before and after the introduction of treatment with topiramate (median dose 300 mg) were compared with changes in test performance of 18 patients who had undergone repeat neuropsychological assessments at the same time intervals. Complaints of cognitive decline precipitated referral for reassessment in five cases in the topiramate treated group. The groups were matched for age and intellectual level at the time of the first assessment. Patients were assessed using the WAIS-R, tests of verbal and non-verbal memory, language, and perceptual processing. A subgroup of patients underwent a brief reassessment after the withdrawal or substantial reduction of topiramate. Repeat assessments in those taking topiramate were associated with a significant deterioration in many domains, which were not seen in the comparison group. The greatest changes were for verbal IQ, verbal fluency, and verbal learning (p<0. 001). Improvements in verbal fluency (p<0.05), verbal learning (p<0. 01), and digit span (p<0.001) were recorded in those patients who had topiramate withdrawn or reduced. In our patient group topiramate had a negative impact on cognition which was consistent with subjective complaints of patients. Tests requiring verbal processing seemed especially sensitive to the drug. A decline in verbal intellect (VIQ), a measure which has been considered by some to be insensitive to antiepileptic drug effects, was particularly striking. Caution is warranted in the interpretation of the findings due to methodological limitations of the study design. Further investigation of mediating factors such as serum concentrations, comedication, and other potential risk factors, however, is needed to enable appropriate targeting of treatment with this effective antiepileptic agent.


Patient and Parent Satisfaction With Sling Use After Pediatric Upper Extremity Fractures: A Randomized Controlled Trial of a Customized Cast-Sling Versus Standard Cast and Sling.
Long arm cast immobilization after operative and nonoperative treatment of pediatric upper extremity fractures is common. The use of a sling to aid in carrying the casted extremity as well as provide further immobilization is also common practice. Off-the-shelf slings vary in quality and fit, can be confusing for parents/patients to apply, and lead to frustration and dissatisfaction with its use. The purpose of this investigation was to compare patient/parent centered outcomes after the use of a customized sling compared with a standard sling by utilizing a prospective, randomized-controlled trial. We hypothesized that patients and their parents would find the "Providence" Pedi Cast-Sling (PPCS) to be more convenient and be more satisfied with its use compare with a standard sling. Eligible subjects included patients 0 to 18 years old, evaluated at an urban, tertiary care pediatric emergency department (ED), who sustained an upper extremity fracture that required placement of a long-arm cast. Exclusion criteria were: open fractures; fractures at multiple levels; fractures requiring urgent/emergent surgery; admitted patients; bivalved casts. Patients were randomized to receive a standard sling or a PPCS. Questionnaires assessing patient/parent satisfaction, preferences, sling-use, and pain level were collected at patients' first follow-up visit. A total of 100 patients were randomized with 71 questionnaires available for analysis (39 standard sling vs. 32 PPCS). There were statistically significant differences for satisfaction scores for all related questions between patients who received a PPCS compared with a standard sling. Patients and their parents were more likely to choose the PPCS (P=0.001), were more satisfied with the PPCS (P<0.006), and reported the PPCS to be more convenient than a standard sling (P<0.001). Patients who received a PPCS wore the sling for a greater number of hours during the day (10.3±7.1 h) compared with those who received a standard sling (5.9±5.0 h) (P=0.004). The "Providence" Pedi Cast-Sling is a superior alternative to an off-the-shelf sling when used after the application of a long-arm cast providing greater convenience, compliance, and satisfaction to both patients and families. Level I-Prospective randomized-controlled trial.


Variation in the thermal ecology of an endemic iguana from Mexico reduces its vulnerability to global warming.
The persistence of reptile populations in a specific location is influenced by individuals' capacity to regulate their body temperatures, among other factors. Anthropogenic climate change may pose a risk to the survival of ectothermic animals due to their dependence on external heat sources to thermoregulate. In this study, we calculated indices of thermal habitat quality, thermoregulatory precision, and thermoregulatory effectiveness for the endemic spiny-tailed iguana Ctenosaura oaxacana. We evaluated these indices and the thermoregulatory behavior of the iguanas in the four types of vegetation that provide the most favorable conditions for thermoregulation. We also performed our experiments during both the wet and dry seasons to capture the full range of thermal conditions available to C. oaxacana over the course of a year. Finally, we evaluated the potential niche for the iguana in the years 2020, 2050, and 2080. Thermoregulation depends on both seasonal and environmental factors in this species. We found that thermoregulation effectiveness in both wet and dry seasons depends not only on the thermal conditions of the immediate environment, but also on the cover vegetation and habitat structure available across the range of habitats the species uses. Thus, heterogeneous habitats with dispersed vegetation may be most suitable for this species' thermoregulatory strategy. Likewise, niche modeling results suggested that suitable habitat for our study species may continue to be available for the next few decades, despite global warming tendencies, as long as cover vegetation remains unaltered. Our results suggest that thermoregulation is a complex process that cannot be generalized for all ectothermic species inhabiting a given region. We also found that temperature changes are not the only factor one must consider when estimating the risk of species loss. To understand the necessary thermal conditions and extinction risk for any ectothermic species, it is necessary to focus studies on the species' general ecology.


Experimental infection of coho salmon Oncorhynchus kisutch by exposure of skin, gills and intestine with Piscirickettsia salmonis.
Piscirickettsiosis pathogenesis was examined using some tissues as entry portals of Piscirickettsia salmonis in coho salmon. Juvenile fish, weighing approximately 8.4 g, were used in this trial. Inocula were prepared using the strain SLGO-95 of P. salmonis. The micro-organism was cultured in the CHSE-214 cell line as described by Fryer et al. (1990) and doses containing 10(4.7) and 10(3.7) TCID50 were prepared. Each dose was used to infect the fish via skin, gills and intestine. Skin and gills were exposed by calibrated drops, and the intestine by an intubation through the anal opening. Some fish were injected intraperitoneally with the same P. salmonis doses, as positive virulence controls. Sham-inoculated fish for each of the tested routes were also included as negative controls. Piscirickettsiosis was experimentally reproduced with all the inoculation methods. Cumulative mortalities and survival analyses showed that the most effective entry portal was skin followed by intestinal intubation and finally by gill infection.


Long-term memory for odors: influences of familiarity and identification across 64 days.
Few studies have investigated long-term odor recognition memory, although some early observations suggested that the forgetting rate of olfactory representations is slower than for other sensory modalities. This study investigated recognition memory across 64 days for high and low familiar odors and faces. Memory was assessed in 83 young participants at 4 occasions; immediate, 4, 16, and 64 days after encoding. The results indicated significant forgetting for odors and faces across the 64 days. The forgetting functions for the 2 modalities were not fundamentally different. Moreover, high familiar odors and faces were better remembered than low familiar ones, indicating an important role of semantic knowledge on recognition proficiency for both modalities. Although odor recognition was significantly better than chance at the 64 days testing, memory for the low familiar odors was relatively poor. Also, the results indicated that odor identification consistency across sessions, irrespective of accuracy, was positively related to successful recognition.


[Pulmonary symptoms of primary immunodeficiency diseases].
Patients who lack major components of the immune system carry an increased risk for severe and recurrent pulmonary infections at those respiratory sites were the deficient component would, in the normal state, have its greatest concentration. We report different pulmonary manifestations in pediatric patients with primary immunodeficiency disease (PID). We studied 44 children younger than 14 years old, who were diagnosed of PID in our pediatric department between January 1990 and May 1996. Antibody deficiencies were the most frequent disorders (27/44; 61.3%) followed by PID associated with or secondary to other disorders (10/44; 22.7%) and defects of phagocyte function (5/44; 11.3%). Twenty-seven patients (61.3%) showed relevant pulmonary manifestations that required assistance in the division of pediatric pulmonology. Bronchial responsiveness was seen in 17/27, 11/27 had recurrent pneumonias with development of bronchiectasis in 7/27. Opportunistic or severe pneumonias leading to acute respiratory failure were diagnosed in 9/27. Necrotizing pneumonias leading to development of pneumatoceles, cavities or abscesses was seen in 3/27 with the same rate for lymphoid interstitial pneumonia. Respiratory symptoms were the first manifestations of PID in 19/27 (70.3%). The findings of the study emphasize the responsibility of the pediatric pulmonologists in avoiding the delayed diagnosis of PID since the prognosis depends on the precocity of diagnosis.


Immunoadsorption of Cryptococcus-specific suppressor T-cell factors.
In the murine cryptococcal suppressor cell circuit, two different T-cell suppressor factors, TsF1 and TsF2, have been identified which specifically suppress the delayed-type hypersensitivity (DTH) response to cryptococcal culture filtrate antigen (CneF). TsF1 is produced by a first-order T suppressor (Ts1) cell population and suppresses the afferent limb of the DTH response, whereas TsF2 is produced by a second-order T suppressor (Ts2) cell population and suppresses the efferent limb of the cryptococcal DTH response. The objective of this study was to ascertain whether TsF1 or TsF2 could bind to cryptococcal antigen. To assess this, adsorption of TsF1 and TsF2 was performed with heat-killed Cryptococcus neoformans cells and by solid-phase immunoadsorption (SPIA) on columns containing cryptococcal antigens, i.e., CneF covalently bound to Sepharose 4B. The suppressive effect of TsF1 was removed by adsorption with intact heat-killed cryptococci and by SPIA on CneF-Sepharose 4B. The binding of cryptococcal TsF1 to the cryptococcal SPIA column was shown to be specific since Sepharose 4B columns either coupled with Saccharomyces cerevisiae mannan or blocked with glycine did not adsorb the suppressor activity. In contrast, the suppressive component of TsF2 did not bind to heat-killed cryptococci, CneF-Sepharose 4B, S. cerevisiae mannan-Sepharose 4B, or glycine-Sepharose 4B columns. These results, together with the finding that cryptococcal antigen, anticryptococcal antibody, and C1q-binding immune complexes were not demonstrated in either TsF1 or TsF2, establish that TsF1 and TsF2 can be differentiated on the basis of their affinity for cryptococcal antigen.


Mechanisms of myopia in Cohen syndrome mapped to chromosome 8q22.
To analyze the mechanisms of myopia in Cohen syndrome (Mendelian Inheritance in Man [MIM] no. 216550). A cross-sectional study of 22 Finnish patients (age range, 2-57 years) with Cohen syndrome, which maps to chromosome 8q22, was undertaken to record cycloplegic refraction, keratometry (corneal power and radius of curvature), biometry (anterior chamber depth [ACD], lens thickness [LT], axial [AL] and vitreal length [VL]), and Hoffer Q-modeled lens power. These components of refraction were correlated to age and spherical equivalent (SE) at the corneal plane. Contribution to total myopia of refractive (corneal and lenticular) and axial components was modeled by multiple linear regression and by estimating the effect of deviation from population mean values. The mean SE in patients with Cohen syndrome older than 10 years was -9.35 D; the mean cylinder power, +1.70 D; and the mean anisometropia, 0.53 D. Relative to the emmetropic eye of a young adult, the AL and VL (mean, 23.9 and 16.6 mm, respectively) and lens power (mean, 30.30 D) were higher in 74% and 93% of patients, respectively, and the ACD (mean, 2.5 mm) was smaller and the LT (mean, 4.9 mm) and corneal power (mean, 45.63 D) higher than average in all patients. Corneal power (r = 0.513, P = 0.021) increased with age, but AL and VL (P = 0.46 and 0.54, respectively) and lens power (P = 0.89) did not correlate with age. The lens power decreased with AL (r = -0.564, P = 0.029) and tended to increase with corneal power (r = 0.475, P = 0.074). Multiple linear regression identified AL and corneal power as independent predictors of SE. Based on deviation from population means, the lens power explained 55%, corneal power 23%, and AL 22% of total myopia. ACD decreased and LT increased markedly with age, rendering angle-closure glaucoma a possibility. Myopia in Cohen syndrome is mainly refractive in type and is due to high corneal and lenticular power, which is otherwise rare in young patients. It may be superimposed on axial myopia, probably related to polygenic factors that determine myopia in the general population. The refractive myopia in Cohen syndrome may result from dysgenesis and atrophy of the cornea, ciliary body, and iris, which in turn cause iridial and zonular laxity and spherophakia.


Hydrogen Sulfide Inhibits High Glucose-Induced sFlt-1 Production via Decreasing ADAM17 Expression in 3T3-L1 Adipocytes.
Hydrogen sulfide (H2S) has recently been identified as an endogenous gaseous signaling molecule. The aim of the present study was to investigate the effect of H2S on high glucose- (HG-) induced ADAM17 expression and sFlt-1 production in 3T3-L1 adipocytes. Firstly, we found that HG DMEM upregulated the expression of ADAM17 and production of sFlt-1 in 3T3-L1 adipocytes. Knocking down ADAM17 attenuated the effect of high glucose on sFlt-1 production in adipocytes. HG decreased the expression of CSE and 3-MST, as well as the endogenous H2S production. Furthermore, knocking down CSE and 3-MST significantly increased ADAM17 expression and sFlt-1 production. The addition of exogenous H2S through the administration of sodium hydrosulfide (NaHS) inhibited HG-induced upregulation of ADAM17 expression and sFlt-1 production. In conclusion, decreased expression of CSE and 3-MST and the subsequent decrease in H2S production contribute to high glucose-induced sFlt-1 production via activating ADAM17 in adipocytes. Exogenous H2S donor NaHS has a potential therapeutic value for diabetic vascular complications.


The complementary nature of query-based and directed health information exchange in primary care practice.
Many policymakers and advocates assume that directed and query-based health information exchange (HIE) work together to meet organizations' interoperability needs, but this is not grounded in a substantial evidence base. This study sought to clarify the relationship between the usage of these 2 approaches to HIE. System user log files from a regional HIE organization and electronic health record system were combined to model the usage of HIE associated with a patient visit at 3 federally qualified health centers in New York. Regression models tested the hypothesis that directed HIE usage was associated with query-based usage and adjusted for factors reflective of the FITT (Fit between Individuals, Task & Technology) framework. Follow-up interviews with 8 key informants helped interpret findings. Usage of query-based HIE occurred in 3.1% of encounters and directed HIE in 23.5%. Query-based usage was 0.6 percentage points higher when directed HIE provided imaging information, and 4.8 percentage points higher when directed HIE provided clinical documents. The probability of query-based HIE was lower for specialist visits, higher for postdischarge visits, and higher for encounters with nurse practitioners. Informants used query-based HIE after directed HIE to obtain additional information, support transitions of care, or in cases of abnormal results. The complementary nature of directed and query-based HIE indicates that both HIE functionalities should be incorporated into EHR Certification Criteria. Quantitative and qualitative findings suggest that directed and query-based HIE exist in a complementary manner in ambulatory care settings.


Plasma homocysteine, B vitamins, and amino acid concentrations in cats with cardiomyopathy and arterial thromboembolism.
Arterial thromboembolism (ATE) is a common complication of cats with cardiomyopathy (CM), but little is known about the pathophysiology of ATE. In people, high plasma concentrations of homocysteine and low B vitamin concentrations are risk factors for peripheral vascular disease. In addition, low plasma arginine concentrations have been linked to endothelial dysfunction. The purpose of this study was to compare concentrations of homocysteine, B vitamins, and amino acids in plasma of normal cats to those of cats with CM and ATE. Plasma concentrations of homocysteine, vitamin B6, vitamin B12, folate, and amino acids were measured in 29 healthy cats, 27 cats with CM alone, and 28 cats with both CM and ATE. No differences were found between groups in homocysteine or folate. Mean vitamin B12 concentration (mean +/- standard deviation) was lower in cats with ATE (866 +/- 367 pg/mL) and cats with CM (939 +/- 389 pg/mL) compared with healthy controls (1,650 +/- 700 pg/mL; P < .001). Mean vitamin B6 concentration was lower in cats with ATE (3,247 +/- 1.215 pmol/mL) and cats with CM (3,200 +/- 906 pmol/mL) compared with healthy control animals (4,380 +/- 1,302 pmol/mL; P = .005). Plasma arginine concentrations were lower in cats with ATE (75 +/- 33 nmol/mL) compared with cats with CM (106 +/- 25 nmol/mL) and healthy control animals (96 +/- 25 nmol/ mL; P < .001). Vitamin B12 concentration was significantly correlated with left atrial size. We interpret the results of this study to suggest that vitamin B12 and arginine may play a role in CM and ATE of cats.


Pseudomalignant cutaneous amoebiasis.
Two cases of cutaneous amoebiasis which presented as malignant ulcers are reported. In non-healing ulcers in the perianal or gluteal region, the possibility of amoebiasis should be kept in mind, especially in the tropics. An immediate examination of the scraping from the ulcer is sufficient to confirm or rule out the diagnosis.


A deficiency of uPAR alters endothelial angiogenic function and cell morphology.
The angiogenic potential of a cell requires dynamic reorganization of the cytoskeletal architecture that involves the interaction of urokinase-type plasminogen activator receptor (uPAR) with the extracellular matrix. This study focuses on the effect of uPAR deficiency (uPAR-/-) on angiogenic function and associated cytoskeletal organization. Utilizing murine endothelial cells, it was observed that adhesion, migration, proliferation, and capillary tube formation were altered in uPAR-/- cells compared to wild-type (WT) cells. On a vitronectin (Vn) matrix, uPAR-/- cells acquired a "fried egg" morphology characterized by circular actin organization and lack of lamellipodia formation. The up-regulation of β1 integrin, FAK(P-Tyr925), and paxillin (P-Tyr118), and decreased Rac1 activation, suggested increased focal adhesions, but delayed focal adhesion turnover in uPAR-/- cells. This accounted for the enhanced adhesion, but attenuated migration, on Vn. VEGF-enriched Matrigel implants from uPAR-/- mice demonstrated a lack of mature vessel formation compared to WT mice. Collectively, these results indicate that a uPAR deficiency leads to decreased angiogenic functions of endothelial cells.


The conformation of the mature dimeric human immunodeficiency virus type 1 RNA genome requires packaging of pol protein.
The packaging of a mature dimeric RNA genome is an essential step in human immunodeficiency virus type 1 (HIV-1) replication. We have previously shown that overexpression of a protease (PR)-inactive HIV-1 Gag-Pro-Pol precursor protein generates noninfectious virions that contain mainly monomeric RNA (M. Shehu-Xhilaga, S. M. Crowe, and J. Mak, J. Virol. 75:1834-1841, 2001). To further define the contribution of HIV-1 Gag and Gag-Pro-Pol to RNA maturation, we analyzed virion RNA dimers derived from Gag particles in the absence of Gag-Pro-Pol. Compared to wild-type (WT) dimeric RNAs, these RNA dimers have altered mobility and low stability under electrophoresis conditions, suggesting that the HIV-1 Gag precursor protein alone is not sufficient to stabilize the dimeric virion RNA structure. The inclusion of an active viral PR, without reverse transcriptase (RT) and integrase (IN), rescued the stability of the virion RNA dimers in the Gag particles but did not restore the mobility of the RNAs, suggesting that RT and IN are also required for virion RNA dimer maturation. Thin-section electron microscopy showed that viral particles deficient in RT and IN contain empty cone-shaped cores. The abnormal core structure indicates a requirement for Gag-Pro-Pol packaging during core maturation. Supplementing viral particles with either RT or IN via Vpr-RT or Vpr-IN alone did not correct the conformation of the dimer RNAs, whereas expression of both RT and IN in trans as a Vpr-RT-IN fusion restored RNA dimer conformation to that of the WT virus and also restored the electron-dense, cone-shaped virion core characteristic of WT virus. Our data suggest a role for RT-IN in RNA dimer conformation and the formation of the electron-dense viral core.


Blunt trauma to the common femoral artery.
A patient with blunt trauma to the groin presented with pain and swelling and was noted to have a bruit and thrill over his right femoral artery. Arteriogram revealed a tear of the endothelium of the common femoral artery. Nonoperative therapy was initiated with continued improvement such that repeat arteriogram 2 months later showed only minimal intimal irregularity, and at 1 year postinjury the patient was asymptomatic.


Hospitals' newest challenge: designing in quality.
The job of producing high-quality products is even more difficult for health care providers than it is for those in manufacturing, where the quality movement began. As a part of the service industry, health care providers are in the position of producing products and delivering services at the moment of sale. Our task is to improve the quality of all of these simultaneous and interrelated processes. Traditionally, health care providers have made efforts to improve their products and services without realizing the impact that could be made by also improving resources, processes, and outcomes. This article is an overview of the new direction we have been taking: Retrospective review. Critical pathways. Building quality into all areas (resources, processes, products and services, and outcomes). Focused study of outcomes). We foresee a further evolution that will lead to exciting new methods for understanding and delivering high-quality care.


Addiction research centres and the nurturing of creativity. National institute on alcohol and drugs policies, Brazil.
The National Institute of Public Policy for Alcohol and Other Drugs (INPAD) is based at the Federal University of São Paulo, Brazil, and was created to collect scientific evidence regarding epidemiology, develop new therapeutic approaches, study health economics and provide education to subsidize the proper measures to change the Brazilian scenario of alcohol and drug consumption. Policies directed towards the control of alcohol and drugs in Brazil are fragmented, poorly enforced and therefore ineffective. The unregulated market of alcohol in Brazil has contributed to the worsening health of the Brazilian population. Since 1994, INPAD has participated actively in academic debates and discussions about alcohol and drug policies and their effects on the political welfare of the country. Many scientific papers and books have been published on this subject, and the internet and other media have provided excellent opportunities for the dissemination of specialized information to the general population.


Variation of bispectral index in children aged 1-12 years under propofol anesthesia: an observational study.
The use of the bispectral index (BIS) is widespread in pediatric anesthesia, but few studies have attempted to perform a detailed evaluation of how BIS varies according to age in children under propofol anesthesia. This prospective study aimed to explore the exact relationship between BIS value and the age of 1- to 12-year-old children under propofol anesthesia. This study enrolled 165 children (1 < yr. ≤ 12), scheduled for surgery under anesthesia, and divided them into 11 age groups. Of the 165 participants, 157 completed the study protocol. All patients were anesthetized with propofol for over 30 s. An observation period of 4 min followed. BIS values were recorded at 0 (immediately after propofol injection), 30, 40, 50, 60, 90, 120, 180, and 240 s after the injection. BIS values at each time point corresponding to the 11 age groups were compared using repeated measures ANOVA. BIS values significantly differed among the nine time points (p < 0.01) as well as among the different age groups (p < 0.01) after propofol administration. Post-hoc Bonferroni tests showed a difference in BIS values between groups 1-4 (1 < yr. ≤ 5) and groups 5-11(5 < yr. ≤ 12). BIS values were lower in the latter than in the former, from 50 to 240 s. The minimum BIS values in group 1 < yr. ≤ 5 and in group 5 < yr. ≤ 12 were recorded at 60 s as 49 ± 17 and 35 ± 14, respectively. During propofol anesthesia, the BIS values were closely related to age, which can be divided into two groups: 1 < yr. ≤ 5 and 5 < yr. ≤ 12. BIS values of the older age group were lower than those of the younger age group at the same time points. Registration number: chictr-roc-16008630 . Registered on 12 June 2016. Retrospectively registered.


Activation of circulated immune cells and inflammatory immune adherence are involved in the whole process of acute venous thrombosis.
To investigate localization and distribution of integrin subunit β1, β2 and β3 and morphological changes of ligand-recepter binding in thrombi of acute pulmonary embolism (PE) patients and explore activation of circulated immune cells, inflammatory immune adherence and coagulation response in acute venous thrombosis. Thrombi were collected from patients with acute PE. Immunohistochemistry was done to detect the expression and distribution of integrin β1, β2 and β3 in cells within thrombi, and ligands of integrin subunit β1, β2 and β3 were also determined by immunohistochemistry within the thrombi. 1) Acute venous thrombi were red thrombi composed of skeletons and filamentous mesh containing large amounts of red blood cells and white blood cells; 2) Integrin subunit β1, β2 and β3 were expressed on lymphocytes, neutrophils and platelets; 3) No expression of integrin β1 ligands: Laminin, Fibronectin, Collagen I or Collagen-II on lymphocytes; integrin β2 ligands including ICAM, factor X and iC3b are distributed on neutrophils, and ligand fibrinogen bound to neutrophils; integrin β3 was expressed on platelets which form the skeleton of thrombi and bound to fibrinogen to construct mesh structure; 4) Factor Xa was expressed on the filamentous mesh; 5) Filamentous mesh was fully filled with red blood cell dominant blood cells. Acute venous thrombosis is an activation process of circulated lymphocytes, neutrophils and platelets mainly, and a whole process including integrin subunit β2 and β3 binding with their ligands. Activation of immune cells, inflammatory immune adherence and coagulation response are involved in the acute venous thrombosis.


Structural determinants of factor IX(a) binding in nitrophorin 2, a lipocalin inhibitor of the intrinsic coagulation pathway.
Nitrophorin 2 (NP2) is a salivary lipocalin from Rhodnius prolixus that binds with coagulation factors IX (fIX) and IXa (fIXa). Binding of NP2 with fIXa results in potent inhibition of the intrinsic factor Xase complex. A panel of site-directed surface mutants of NP2 was generated to locate determinants of high affinity fIX(a) binding. The locations of the mutations were based on comparisons with the related, but less potent, inhibitor nitrophorin 3 (NP3). Three point mutants (K21A, K92A, and V94A) were found that clearly reduced the inhibitory potency as measured by the activity of a reconstituted factor Xase system. Binding of NP2 with fIXa and fIX as measured by surface plasmon resonance and isothermal titration calorimetry was reduced in a similar manner. Of the three mutants, two (K92A and V94A) were located on the loop connecting beta-strands E and F of the lipocalin beta-barrel. The largest changes were seen with the K92A mutation, which lies at the apex of the loop, with a smaller effect being seen with mutation of Val(94). Combination of four E-F loop mutations (K92A, A93K, V94A, E97A) in a single mutant reduced the inhibitory potency and binding to levels similar to those seen with NP3 without affecting heme or histamine binding.


LRRC8A anion channels modulate vascular reactivity via association with myosin phosphatase rho interacting protein.
Leucine-rich repeat containing 8A (LRRC8A) volume regulated anion channels (VRACs) are activated by inflammatory and pro-contractile stimuli including tumor necrosis factor alpha (TNFα), angiotensin II and stretch. LRRC8A associates with NADPH oxidase 1 (Nox1) and supports extracellular superoxide production. We tested the hypothesis that VRACs modulate TNFα signaling and vasomotor function in mice lacking LRRC8A exclusively in vascular smooth muscle cells (VSMCs, Sm22α-Cre, Knockout). Knockout (KO) mesenteric vessels contracted normally but relaxation to acetylcholine (ACh) and sodium nitroprusside (SNP) was enhanced compared to wild type (WT). Forty-eight hours of ex vivo exposure to TNFα (10 ng/mL) enhanced contraction to norepinephrine (NE) and markedly impaired dilation to ACh and SNP in WT but not KO vessels. VRAC blockade (carbenoxolone, CBX, 100 μM, 20 min) enhanced dilation of control rings and restored impaired dilation following TNFα exposure. Myogenic tone was absent in KO rings. LRRC8A immunoprecipitation followed by mass spectroscopy identified 33 proteins that interacted with LRRC8A. Among them, the myosin phosphatase rho-interacting protein (MPRIP) links RhoA, MYPT1 and actin. LRRC8A-MPRIP co-localization was confirmed by confocal imaging of tagged proteins, Proximity Ligation Assays, and IP/western blots. siLRRC8A or CBX treatment decreased RhoA activity in VSMCs, and MYPT1 phosphorylation was reduced in KO mesenteries suggesting that reduced ROCK activity contributes to enhanced relaxation. MPRIP was a target of redox modification, becoming oxidized (sulfenylated) after TNFα exposure. Interaction of LRRC8A with MPRIP may allow redox regulation of the cytoskeleton by linking Nox1 activation to impaired vasodilation. This identifies VRACs as potential targets for treatment or prevention of vascular disease.


Person-centered risk assessment framework: assessing and managing risk in older adults living with dementia.
This study pilot-tested the person-centered risk assessment framework (PCRAF), a framework for managing risk among persons living with dementia (PLWD) in primary care. Healthcare providers (N = 7) piloting the PCRAF completed a survey, rating their satisfaction with the tool, and an interview to gather their perceptions of the PCRAF. PLWD and care partners (N = 12) completed a survey, rating their satisfaction with safety planning. Care providers were very satisfied with the tool; however, patient or care partner inability to perceive or understand safety risks was a challenge. Use of the PCRAF was perceived as an opportunity to empower self-management, gather PLWD and care partner perspectives, reduce burden for care partners and increase understanding of potential risks. Patients and care partners were very satisfied with the way in which they were included in the risk discussion. The PCRAF is a promising new tool to reduce risks associated with dementia.


Recurrent chemical meningitis in craniopharyngioma without reduction in size of cyst: case report of two cases and review of the literature.
Chemical meningitis is a rare phenomenon due to rupture of a craniopharyngioma cyst; it develops because of presence of cholesterol crystals in cyst fluid secreted by the squamous epithelium lining of the cyst. Spontaneous rupture of the cyst may present with or without meningitis, depending upon the cholesterol contents of cystic fluid of tumor. A decrease in the size of cyst may or may not be noted. Here we report two cases of craniopharyngioma, one which had a recurrent episode of chemical meningitis after surgery due to leak from residual component of the craniopharyngioma without evidence of recurrence of lesion, or decrease in the size of the cyst. Another case presented with initial symptoms and signs of meningitis, with no change in the size of the lesion after the second episode of chemical meningitis.


Stress Echo 2030: The Novel ABCDE-(FGLPR) Protocol to Define the Future of Imaging.
With stress echo (SE) 2020 study, a new standard of practice in stress imaging was developed and disseminated: the ABCDE protocol for functional testing within and beyond CAD. ABCDE protocol was the fruit of SE 2020, and is the seed of SE 2030, which is articulated in 12 projects: 1-SE in coronary artery disease (SECAD); 2-SE in diastolic heart failure (SEDIA); 3-SE in hypertrophic cardiomyopathy (SEHCA); 4-SE post-chest radiotherapy and chemotherapy (SERA); 5-Artificial intelligence SE evaluation (AI-SEE); 6-Environmental stress echocardiography and air pollution (ESTER); 7-SE in repaired Tetralogy of Fallot (SETOF); 8-SE in post-COVID-19 (SECOV); 9: Recovery by stress echo of conventionally unfit donor good hearts (RESURGE); 10-SE for mitral ischemic regurgitation (SEMIR); 11-SE in valvular heart disease (SEVA); 12-SE for coronary vasospasm (SESPASM). The study aims to recruit in the next 5 years (2021-2025) ≥10,000 patients followed for ≥5 years (up to 2030) from ≥20 quality-controlled laboratories from ≥10 countries. In this COVID-19 era of sustainable health care delivery, SE2030 will provide the evidence to finally recommend SE as the optimal and versatile imaging modality for functional testing anywhere, any time, and in any patient.


Integrated behavioral treatments for comorbid anxiety and substance use disorders: A model for understanding integrated treatment approaches and meta-analysis to evaluate their efficacy.
Substance use disorders (SUD) and anxiety disorders are highly comorbid, and this comorbidity is associated with poor clinical outcomes. Emerging research in the last decade has shifted from addressing these problems separately to the development and evaluation of behavioral treatments that integrate care for anxiety disorders (or elevated symptoms of anxiety) and SUD. An extensive literature search revealed a sufficient number of studies (K=11) to conduct a meta-analysis comparing the efficacy of integrated SUD/anxiety disorder behavioral treatment to SUD treatment alone on substance use and anxiety symptom outcomes. Randomized clinical trials including those with SUD and either anxiety disorders or elevations in constructs implicated in the maintenance of anxiety disorder/SUD comorbidity were included. This study meta-analyzes the effects of these studies. Integrated treatments outperformed SUD treatments alone on both substance use and anxiety outcomes, with small to moderate effects favoring integrated treatments. There was no significant heterogeneity across studies in the primary analyses, such that moderator analyses to identify variables that yielded differential patterns of effect sizes were not conducted. Integrated treatments for SUD/anxiety disorders demonstrate an incremental but significant and clinically meaningful improvement over SUD treatment alone for SUD/anxiety disorder comorbidity. Implications for future research and clinical practice paradigm shifting are discussed.


Effect and mechanism of aqueous extract of Chinese herbal prescription (TFK) in treating gout arthritis.
With the rapid development of China's economic level, great changes have taken place in people's diet structure, gout has become a common disease that puzzles people's health, seriously affects the realization of China's "Healthy China" strategic goal. Gouty arthritis (GA) is a common joint disease caused by chronic purine metabolism disorder. Currently, drugs used to treat GA are allopurinol and colchicine. However, these drugs can only temporarily relieve the clinical symptoms of GA with significant side effects. More and more basic and clinical studies have confirmed that Traditional Chinese medicine has definite curative effect on GA. To elucidate the potential molecular mechanism of Tongfengkang (TFK) in the treatment of GA, and to provide experimental basis for the search and development of efficient and low-toxicity Chinese medicine for GA treatment. Aqueous extract of TFK (AETFK) were determined by liquid phase high resolution mass spectrometry and the possible effective constituents were screened out. Acute GA model rats were established to detect the anti-inflammatory and detumification effects of AETFK on GA and explore the potential mechanism. The effect of AETFK on serum uric acid and urinary uric acid levels in acute GA rats was determined by automatic biochemical analyzer, and the effect of AETFK on the expression of acute GA-related immunoinflammatory factors were determined by protein thermal fluorescence chip. The effect of AETFK on the concentration of neutrophils in the joint fluid of acute GA rats were determined by Reichs-Giemsa staining. The effect of AETFK on macrophage activation was detected by ELISA. In order to further investigate the mechanism of AETFK in the treatment of GA, a rat model of hyperuricemia was established to detect the effect of AETFK on the level of uric acid in hyperuricemia model rats. Biochemical indexes of liver and kidney and hematoxylin-eosin staining (HE) were used to evaluate the effects of AETFK on the organs, and to preliminatively evaluate the safety of ventilation confufang. Compared with the model group, the joint swelling degree of GA rats in AETFK treatment group were significantly reduced, and the levels of blood uric acid and urine uric acid were also significantly decreased. Protein thermal fluorescence microarray results showed that the levels of gout - related inflammatory factors in GA rats in AETFK treatment group were significantly lower than those in control group. Reichsen-giemsa staining and ELISA showed that AETFK could reduce the activation of macrophages and the accumulation of neutrophils in the joint fluid. The results of liver and kidney biochemical indexes and HE staining showed that no obvious tissue damage was observed in the organs of rats treated with AETFK. AETFK not only has significant anti-inflammatory effects on GA, but also can significantly reduce the level of blood uric acid in GA rats, without obvious toxic and side effects. These effects may be related to AETFK's inhibition of neutrophil enrichment and macrophage activation during early inflammation.


[Hallux valgus: which treatment].
Hallux valgus deformity is characterized by lateral deviation of the proximal phalanx on the metatarsal head. Variable severity of the deviation causes prominence of the medial eminence, giving rise to a painful bunion deformity. Many patients will find permanent or temporary relief with non surgical measures. For those who failed conservative treatment, surgery may cure their symptoms. However unsatisfactory outcomes following hallux valgus surgery is not rare. Therefore a clear understanding of the pathology and of the patient's expectations must anticipate any treatment strategy in order to optimise patient's outcome. Cosmetic considerations should never be an indication for surgery.


Risk of bacteremia after endotracheal intubation for general anesthesia.
Blood cultures were obtained before and after endotracheal intubation to assess the risk of bacteremia associated with this procedure and to evaluate the need for prophylactic antibiotics to prevent bacterial endocarditis in patients with structural heart disease requiring general anesthesia. Blood cultures were obtained immediately before intubation and two and ten minutes after intubation in 50 individuals without evidence of structural heart disease who required general anesthesia for elective surgery. Of 32 who had orotracheal intubation, only one demonstrated postintubation bacteremia; of 18 individuals who had nasotracheal intubation, none had evidence of bacteremia. These data suggest that in healthy individuals undergoing endotracheal intubation under direct vision the risk of bacteremia is quite small (about 2%).


In silico determination of transposon-derived miRNAs and targets in Aegilops species.
Transposable elements (TEs) are found almost in all living organism, shaping organisms' genomes. miRNAs are noncoding RNA types which are especially important in gene expression regulations. Many previously determined plant miRNAs are identical/homologous to transposons (TE-MIR). The aim of this study was computational characterization of novel TE-related miRNAs and their targets in Aegilops genome by using stringent criteria. Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses were performed by BLAST2GO. Seventeen novel TE-related miRNAs in Aegilops genome were identified for the first time. GO analyses indicated that 40 targets played different roles in biological processes, cellular components and molecular functions. Moreover, these genes were involved in 10 metabolic pathways such as purine metabolism, nitrogen metabolism, oxidative phosphorylation, etc. as a result of KEGG analyses. Identification of miRNAs and their targets are significant to understand miRNA-TEs relationships and even how TEs affect plant growth and development. Obtaining results of this study are expected to provide possible new insight into Aegilops and its related species, wheat, with respect to miRNAs evolution and domestication.Communicated by Ramaswamy H. Sarma.


Pre- and postnatal exposure to multiple ambient air pollutants and child behavioral problems at five years of age.
Ambient air pollution is emerging as a risk factor for adverse neurological symptoms and early childhood diseases. This study aimed to evaluate the association between pre- and postnatal exposure to air pollutants and childhood behavior by using MOCEH prospective birth cohort data. In total, 353 mother-child pairs at birth, who completed child behavioral assessments using the Korean version of the Child Behavior Checklist at five years of age, were included in the study. Multivariate linear regression (MLR) for single pollutant and Bayesian kernel machine regression (BKMR) for multiple pollutants were conducted. MLR analysis showed that air pollutant exposures during the first trimester were significantly associated with the internalizing problems score after adjusting for covariates. The estimates were 0.19 (0.05-0.32) per 1 μg/m3 increase in PM2.5, 0.13 (0.04-0.22) per 1 μg/m3 increase in PM10, and 0.20 (0.02-0.37) per 1 ppb increase in NO2. The BKMR model analysis revealed that the overall effects of multiple air pollutants during the first trimester of pregnancy and 0-6 months of the infantile period were significantly associated with behavioral problems. Boys showed a stronger associations than girls. Taken together, these results showed that the first trimester of pregnancy and 0-6 months of the infantile period were important for air pollutant exposure because exposure at these periods was associated with behavioral problems in 5-year-old children. Future efforts are required to control air pollution levels and reduce the health burden of vulnerable populations, including pregnant women and children.


Maintenance of phenotype and function of cryopreserved bone-derived cells.
The emerging fields of tissue engineering and regenerative medicine require large numbers of cells for therapy. Although the properties of cells obtained from a variety of fresh tissues have been delineated, the knowledge regarding cryopreserved grafts-derived cells remains elusive. Previous studies have shown that living cells could be isolated from cryopreserved bone grafts. However, whether cryopreserved bone-derived cells can be applied in regenerative medicine is largely unknown. The present study was to evaluate the potential application of cryopreserved grafts-derived cells for tissue regeneration. We showed that cells derived from cryopreserved bone grafts could maintain good proliferation activity and osteogenic phenotype. The biological phenotype of these cells could be well preserved. The transplantation of cryopreserved bone-derived cells on scaffold could promote new bone formation in nude mice and enhance the osteointegration for dental implants in canine, which confirmed their osteogenic capacity, and showed that cells derived from cryopreserved bone were comparable to that of fresh bone in terms of the ability to promote osteogenesis in vivo. This work demonstrates that cryopreserved bone grafts may represent a novel, accessible source of cells for tissue regeneration therapy, and the results of our study may also stimulate the development of other cryopreservation techniques in basic and clinical studies.


Serum Vitamin D Concentrations and Cognitive Function in a Population-Based Study among Older Adults in South Germany.
Our objective was to investigate the associations of vitamin D serum levels with dementia and cognitive function in specific domains in community dwelling older adults. Between 2009 and 2010, we conducted a cross-sectional study in 1,373 individuals (56% men) aged 65+ years in the "Activity and Function in the Elderly in Ulm" (ActiFE) study. Dementia was defined as a Mini-Mental State Examination (MMSE) score ≤ 24. The 25-OHD serum level [ng/mL] was measured by an electrochemilumineszenz immunoassay (ECLIA). Logistic regression models were used to calculate odds ratios (OR)s for cognitive domains (cut-point: 10th percentile) by serum 25-OHD concentrations (both continuously and by cut-point of 20 ng/ml for vitamin D deficiency). Mean age of the study population was 75.6 (SD 6.6) years.We identified 75 participants (43% women) with dementia. 25-OHD concentrations were significantly lower in the participants with dementia compared to persons with a MMSE score >24. We also observed an association of continuous 25-OHD serum concentrations with prevalence of dementia (crude OR 1.05, 95% confidence interval (CI), 1.01-1.08, p-value 0.009) per 1 ng/mL decrease, after adjustment the OR was 1.03, 95% CI, 0.995-1.08 (p-value 0.09). Although vitamin D deficiency was tentatively associated with severity of dementia measured by MMSE (OR 1.35, 95% CI, 0.84-2.19), the association was not statistically significant. However, deficits in specific cognitive domains such as executive functions, wordlist encoding, and visual memory (encoding and recall) were significantly associated with low vitamin D concentration. Our results suggest an association between vitamin D deficiency and cognitive function in specific domains in community dwelling older adults.


Avermectin B2 O-methyltransferase activity in "Streptomyces avermitilis" mutants that produce increased amounts of the avermectins.
The level of activity of avermectin B O-methyltransferase, the enzyme which catalyzes the conversion of avermectin B components to avermectin A components, was analyzed in a series of "Streptomyces avermitilis" mutants selected for increased production of the avermectins. In all of the mutants, increased avermectin production was accompanied by increased avermectin B O-methyltransferase activity. Both the average specific activity and the maximum observed specific activity of avermectin B O-methyltransferase increased in direct proportion to avermectin production. The level of avermectin B O-methyltransferase alone did not determine the extent of conversion of avermectin B components to avermectin A components, since a constant ratio of B components to A components was maintained throughout the fermentation even though avermectin B O-methyltransferase specific activity varied three- to fivefold. These results indicate that avermectin B O-methyltransferase is not rate limiting. The correlation between avermectin B O-methyltransferase specific activity and avermectin production is compatible with the hypothesis that genes coding for successive steps in the same secondary metabolite biosynthetic pathway are coordinately regulated.


Appraising the ANT: Psychometric and theoretical considerations of the Attention Network Test.
The Attention Network Test (ANT) is a tool used to assess the efficiency of the 3 attention networks-alerting, orienting, and executive control. The ANT has become popular in the neuropsychological literature since its first description in 2002, with some form of the task currently appearing in no less than 65 original research papers. Although several general reviews of the ANT exist, none provide an analysis of its psychometric properties. Data from 15 unique studies were collected, resulting in a large sample (N = 1,129) of healthy individuals. Split-half reliability, variance structure, distribution shape, and independence of measurement of the 3 attention network scores were analyzed, considering both reaction time and accuracy as dependent variables. Split-half reliabilities of reaction time based attention network scores were low for alerting (rweighted = .20, CI 95%weighted [.14, .27], Spearman-Brown r = .38) and orienting (rweighted = .32, CI 95%weighted [.26, .38], Spearman-Brown r = .55), and moderate high for executive control (rweighted = .65, CI 95%weighted [.61, .71], Spearman-Brown r = .81). Analysis of the variance structure of the ANT indicated that power to find significant effects was variable across networks and dependent on the statistical analysis being used. Both analysis of variance (significant interaction observed in 100% of 15 studies) and correlational analyses (multiple-significant inter-network correlations observed) suggest that the networks measured by the ANT are not independent. In the collection, analysis and interpretation of any test data, psychometric properties, such as those reported here for the ANT, must be carefully considered.


Critical role of prostate apoptosis response-4 in determining the sensitivity of pancreatic cancer cells to small-molecule inhibitor-induced apoptosis.
Role of prostate apoptosis response-4 (PAR-4) has been well described in prostate cancer. However, its significance in other cancers has not been fully elucidated. For the current study, we selected four pancreatic cancer cell lines (BxPC-3, Colo-357, L3.6pl, and HPAC) that showed differential endogenous expression of PAR-4. We found that nonpeptidic small-molecule inhibitors (SMI) of Bcl-2 family proteins (apogossypolone and TW-37; 250 nmol/L and 1 micromol/L, respectively) could induce PAR-4-dependent inhibition of cell growth and induction of apoptosis. Sensitivity to apoptosis was directly related to the expression levels of PAR-4 (R = 0.92 and R2 = 0.95). Conversely, small interfering RNA against PAR-4 blocked apoptosis, confirming that PAR-4 is a key player in the apoptotic process. PAR-4 nuclear localization is considered a prerequisite for cells to undergo apoptosis, and we found that the treatment of Colo-357 and L3.6pl cells with 250 nmol/L SMI leads to nuclear localization of PAR-4 as confirmed by 4',6-diamidino-2-phenylindole staining. In combination studies with gemcitabine, pretreatment with SMI leads to sensitization of Colo-357 cells to the growth-inhibitory and apoptotic action of a therapeutic drug, gemcitabine. In an in vivo setting, the maximum tolerated dose of TW-37 in xenograft of severe combined immunodeficient mice (40 mg/kg for three i.v. injections) led to significant tumor inhibition. Our results suggest that the observed antitumor activity of SMIs is mediated through a novel pathway involving induction of PAR-4. To our knowledge, this is the first study reporting SMI-mediated apoptosis involving PAR-4 in pancreatic cancer.


Abdominal wall hernias-A local manifestation of systemically impaired quality of the extracellular matrix.
Throughout life, inguinal hernia develops in approximately every fourth man, some of whom develop multiple hernias. If patients at risk of developing multiple hernias could be identified by a serologic biomarker, treatment might be able to be tailored and improved. Evidence suggests that abdominal wall hernia formation is associated with altered collagen metabolism. The aim of this study was to evaluate biomarkers for type IV and V collagen turnover in patients with multiple hernias and control subjects without hernia. Venous blood was collected from 88 men (mean age, 62 years) with a history of more than 3 hernia repairs and 86, age-matched men without hernias. Biomarkers for synthesis of collagen type IV (P4NP) and type V (P5CP) as well as breakdown (C4M and C5M) were measured in serum by validated, solid-phase, competitive assays. Collagen turnover was indicated by the ratio between the biomarker for synthesis and breakdown. Type IV collagen turnover was 1.4-fold increased in patients with multiple hernias compared to control subjects (P < .001), whereas type V collagen turnover was 1.7-fold decreased (P < .001). Diagnostic power of P5CP was 0.83 (95%C.I.:0.77-0.89), P < .001. Patients with multiple hernias exhibit increased turnover of type IV collagen and a decreased turnover of type V collagen, demonstrating systemically altered collagen turnover. Biomarkers for type V collagen turnover may be used to identify patients at risk for or with multiple hernias.


Construction of a cDNA encoding the multifunctional animal fatty acid synthase and expression in Spodoptera frugiperda cells using baculoviral vectors.
A cDNA encoding the 2505-residue multifunctional rat fatty acid synthase has been constructed and expressed as a catalytically active protein in Spodoptera frugiperda (Sf9) cells using Autographa californica nuclear polyhedrosis virus (baculovirus). The 7.5 kb cDNA was engineered by the amplification and sequential splicing together of seven fragments contained in overlapping cDNAs that collectively spanned the entire rat fatty acid synthase coding sequence. The full-length cDNA was cloned into a baculoviral transfer vector and used together with linearized baculoviral DNA to co-transfect Sf9 cells. Recombinant viral clones were purified and identified by Western blotting. The recombinant fatty acid synthase was expressed maximally 2 days after infection of the Sf9 cells, constituting up to 20% of the soluble cytoplasm, and could be conveniently separated from the insect host fatty acid synthase by high-performance anion-exchange chromatography. The catalytic properties of the purified recombinant fatty acid synthase are indistinguishable from those of the best preparations of the natural protein obtained from rat liver. These results indicate that, in the insect cell host, all seven catalytic components of the 2505-residue recombinant fatty acid synthase fold correctly, the acyl-carrier-protein domain is appropriately phosphopantetheinylated post-translationally, and the multifunctional polypeptide forms catalytically competent dimers. Thus the baculoviral system appears to be well suited for the expression of specific fatty acid synthase mutants that can be used to explore the mechanism by which the seven domains of this multifunctional homodimer co-operate in the biosynthesis of fatty acids.


A comparative study of antithrombotic and antiplatelet activities of different fucoidans from Laminaria japonica.
Fucoidans extracted from brown algae represent an intriguing group of natural fucose-enriched sulfated polysaccharide, with excellent anticoagulant, antimetastatic, antiangiogenic and anti-inflammatory activities. In the present study, we compared antithrombotic activities of four fucoidan fractions with different molecular weight and sulfated ester content from Laminaria japonica in an electrical induced arterial thrombosis and their potential mechanism underlying such activity. In vivo middle molecular weight (MMW) fucoidan fractions with molecular weight about 28000 and 35000 exhibited better antithrombotic activity in electrical induced arterial thrombosis than low molecular weight (LMW) fucoidan LF1 and LF2 (Mw 7600 and 3900). Inhibition of arterial thrombosis occurred at dose of 0.1-0.25mg/kg for MMW fucoidans, accompanied with moderate anticoagulant activity and significant decrease of whole blood viscosity and hematocrit. The antithrombotic effects of MMW Fucoidans might be related with promotion of TFPI content and decrease of TXB2 content, without affecting platelet aggregation and 6-keto-PGF1α content in vivo. In contrast, LMW fucoidans showed a correlation among anticoagulant, antiplatelet and antithrombotic effects in vivo. Antithrombotic action of LF1 and LF2 required high dose of 2.5-10mg/kg, concomitantly with anticoagulant activity and specific inhibition of platelet aggregation in vivo. Their antithrombotic effect might be related to their promotion of TFPI and 6-keto-PGF1α, down regulation of TXB2, without affecting hemorheology. These findings suggested that fucoidan fractions with different molecular weight acted on the antithrombotic action by different mechanism. By comparison, highly sulfated fucoidan LF2 with molecular weight of 3900 seemed to be a more suitable choice of antithrombotic drug for its antithrombotic activity accompanied with specific inhibitory activity on platelet aggregation, low anticoagulant activity and low hemorrhagic risk in vivo.


One surgeon's experience with the piggyback versus the standard technique in orthotopic liver transplantation: is one better than the other?
Traditionally, orthotopic liver transplantation has consisted of a total native hepatectomy that included retrohepatic inferior vena cava. The so-called "piggyback" technique was described by Tzakis et al. (2). It consists of a recipient hepatectomy with preservation of the native retrohepatic IVC and subsequent anastomosis of the homograft suprahepatic IVC to a cuff fashioned from the recipient's suprahepatic veins. In this study, a single surgeon's experience with both techniques during the same period of time is discussed to analyze any significant differences in survival, intraoperative blood loss, length of stay in the ICU, and total length of stay in the hospital. Over a three year period, 128 patients were transplanted at the University of Pittsburgh. Of these, 66 patients (51.6%) had a piggyback (PB) operation while the remaining 62 (48.4%) had a "standard" (ST) operation. The actual 6 month survival was 81.8% in the PB group (54/66) and 74.2% in the ST group (46/62), with no statistical difference between the two. The median intraoperative blood usage was 6 units for the PB group versus 10 units for the ST group (p > 0.02). The median ICU length of stay was the same at 4 days, as was the total hospitalization duration, 21 days. The in-hospital deaths were included in the analysis. The piggyback technique has some advantages, including less bleeding and absence of brachial plexus or phrenic nerve injury. Several other important considerations are discussed. In conclusion, the results with the piggyback technique are equivalent to those obtained with the standard approach.


Which level is responsible for gluteal pain in lumbar disc hernia?
There are many different reasons why patients could be experiencing pain in the gluteal area. Previous studies have shown an association between radicular low back pain (LBP) and gluteal pain (GP). Studies locating the specific level responsible for gluteal pain in lumbar disc hernias have rarely been reported. All patients with lumbar disc herniation (LDH) in the Kanghua hospital from 2010 to 2014 were recruited. All patients underwent a lumbar spine MRI to clarify their LDH diagnosis, and patients were allocated to a GP group and a non-GP group. To determine the cause and effect relationship between LDH and GP, all of the patients were subjected to percutaneous endoscopic lumbar discectomy (PELD). A total of 286 cases were included according to the inclusive criteria, with 168 cases in the GP group and 118 cases in the non-GP group. Of these, in the GP group, 159 cases involved the L4/5 level and 9 cases involved the L5/S1 level, while in the non-GP group, 43 cases involved the L4/5 level and 48 cases involved the L5/S1 level. PELD was performed in both groups. Gluteal pain gradually disappeared after surgery in all of the patients. Gluteal pain recrudesced in a patient with recurrent disc herniation (L4/5). As a clinical finding, gluteal pain is related to low lumbar disc hernia. The L4/5 level is the main level responsible for gluteal pain in lumbar disc hernia. No patients with gluteal pain exhibited involvement at the L3/4 level.


Silencing of claudin-11 is associated with increased invasiveness of gastric cancer cells.
Claudins are membrane proteins that play critical roles in tight junction (TJ) formation and function. Members of the claudin gene family have been demonstrated to be aberrantly regulated, and to participate in the pathogenesis of various human cancers. In the present study, we report that claudin-11 (CLDN11) is silenced in gastric cancer via hypermethylation of its promoter region. Levels of CLDN11 methylation and mRNA expression were measured in primary gastric cancer tissues, noncancerous gastric mucosae, and cell lines of gastric origin using quantitative methylation-specific PCR (qMSP) and quantitative reverse transcriptase-PCR (qRT-PCR), respectively. Analyses of paired gastric cancers and adjacent normal gastric tissues revealed hypermethylation of the CLDN11 promoter region in gastric cancers, and this hypermethylation was significantly correlated with downregulation of CLDN11 expression vs. normal tissues. The CLDN11 promoter region was also hypermethylated in all gastric cancer cell lines tested relative to immortalized normal gastric epithelial cells. Moreover, CLDN11 mRNA expression was inversely correlated with its methylation level. Treatment of CLDN11-nonexpressing gastric cancer cells with 5-aza-2'-deoxycytidine restored CLDN11 expression. Moreover, siRNA-mediated knockdown of CLDN11 expression in normal gastric epithelial cells increased their motility and invasiveness. These data suggest that hypermethylation of CLDN11, leading to downregulated expression, contributes to gastric carcinogenesis by increasing cellular motility and invasiveness. A further understanding of the mechanisms underlying the role of claudin proteins in gastric carcinogenesis will likely help in the identification of novel approaches for diagnosis and therapy of gastric cancer.


New paradigms in drug design and discovery.
The new millennium has ushered in an era of science that will revolutionize a great majority of our daily activities. That revolution is being experienced by a growing number of the population who are pushing the average life expectancy closer to the 80-year mark. The primary reason for this increase is the changes we have made in the last 2-3 decades both in how we live our lives as well as how we treat our maladies when they arise. The advent of new techniques in diagnostics and surgery have allowed many to survive debilitating illnesses when their chances would have been slim only a few years ago. In addition, several new therapeutic agents have been developed in the latter part of the 20th century that have improved our quality of life and increased our overall survival time. New medicines to treat cardiovascular, degenerative, infectious, and neoplastic diseases are rapidly being discovered in an effort to further lengthen our lifetimes. The processes used by academic and industrial scientist to discover new drugs has recently experienced a true renaissance with many new and exciting techniques being developed in only the past 5-10 years. In this review, we will attempt to outline these latest protocols that chemists and biomedical scientist are currently employing to rapidly bring new drugs to the clinic.


[Behaviors in response to the tuberculin skin test conversion in medical students from a university in Lima, Peru].
The aim of this study was to determine the behaviors in response to tuberculin skin test (PPD) conversion and the reasons for starting or not starting treatment for latent tuberculosis infection (TILT) among medical students at a university in Lima, Peru. A total of 548 participants completed a questionnaire; of them, 6.7% tested positive on university admission and 11.1% were recent converters. A total of 55.7% did not start TILT and had no explanation. Of the recent converters, most students who did start TILT were >22 years old (p = 0.01) and correctly answered general knowledge questions (p = 0.04). It is important to note that the PPD conversion rate was higher than that reported in the literature and that most students did not follow the treatment prophylaxis because of a lack of information.


SARS-CoV-2 Omicron-B.1.1.529 leads to widespread escape from neutralizing antibody responses.
On 24th November 2021, the sequence of a new SARS-CoV-2 viral isolate Omicron-B.1.1.529 was announced, containing far more mutations in Spike (S) than previously reported variants. Neutralization titers of Omicron by sera from vaccinees and convalescent subjects infected with early pandemic Alpha, Beta, Gamma, or Delta are substantially reduced, or the sera failed to neutralize. Titers against Omicron are boosted by third vaccine doses and are high in both vaccinated individuals and those infected by Delta. Mutations in Omicron knock out or substantially reduce neutralization by most of the large panel of potent monoclonal antibodies and antibodies under commercial development. Omicron S has structural changes from earlier viruses and uses mutations that confer tight binding to ACE2 to unleash evolution driven by immune escape. This leads to a large number of mutations in the ACE2 binding site and rebalances receptor affinity to that of earlier pandemic viruses.


Fluoride for treating postmenopausal osteoporosis.
To assess the efficacy of fluoride therapy on bone loss, vertebral and non-vertebral fractures and side effects in postmenopausal women. We searched Medline, Current Contents and the Cochrane Controlled Trial Registry up to December 1998. Two independent reviewers selected RCTs which met predetermined inclusion criteria. Two reviewers independently extracted data using predetermined forms and assessed the methodological quality of the trials using a validated scale. For dichotomous outcomes, relative risks (RR) were calculated and for continuous outcomes, weighted mean differences (WMD) of percentage change from baseline were calculated. Where heterogeneity existed (determined by a chi-square test) a random effects model was used. Eleven studies (1429 subjects) met the inclusion criteria. The increase in lumbar spine bone mineral density (BMD) was found to be higher in the treatment group than in the control group with a WMD 8.1% (95%CI: 7.15,9.09) after two years of treatment and 16.1%(95%CI: 14.65,17.5) after four years. The RR for new vertebral fractures was not significant at two years [0.87 (95%CI: 0.51,1.46)] or at four years [0.9(95%CI: 0.71,1.14)]. The RR for new non-vertebral fractures was not significant at two years 1.2(95%CI: 0.68,2.1) but was increased at four years in the treated group 1.85(95%CI: 1.36,2.5), especially if used at high doses and in a non slow release form. The RR for gastrointestinal side effects was not significant at two years 2.18(95%CI: 0.86,1.21) but was increased at four years in the treated group 2.18(95%CI: 1.69,4.57) especially if fluoride was used at high doses and in a non slow release form. The number of withdrawals and dropouts was not different between treated and control groups at two and four years. Although fluoride has an ability to increase BMD at lumbar spine, it does not result in a reduction of vertebral fractures. In increasing the dose of fluoride, one increases the risk of non-vertebral fracture and gastrointestinal side effects without any effect on the vertebral fracture rate.


Differential effects of deep cerebellar nuclei inactivation on reaching and adaptive control.
This study examined the effects of selective inactivation of the cerebellar nuclei in the cat on the control of multijoint trajectories and trajectory adaptation to avoid obstacles. Animals were restrained in a hammock and trained to perform a prehension task in which they reached to grasp a small cube of meat from a narrow food well. To examine trajectory adaptation, reaching was obstructed by placing a horizontal bar in the limb's path. Inactivation was produced by microinjection of the GABA agonist muscimol (0.25-1.0 microg in 1 microL saline). Fastigial nucleus inactivation produced a severe impairment in balance and in head and trunk control but no effect on reaching and grasping. Dentate inactivation slowed movements significantly and produced a significant increase in tip path curvature but did not impair reaching and grasping. Selective inactivation of the anterior and posterior interpositus nuclei did not impair grasping but severely decreased the accuracy of reaching movements and produced different biases in wrist and paw paths. Anterior interpositus inactivation produced movement slowing (wrist speed) and under-reaching to the food well. Wrist and tip paths showed anterior biases and became more curved. Also animals could no longer make anticipatory adjustments in limb kinematics to avoid obstructions but sensory-evoked corrective responses were preserved. Posterior interpositus inactivation produced a significant increase in wrist speed and overreaching. Wrist and tip paths showed a posterior bias and became more curved, although in a different way than during anterior interpositus inactivation. Posterior interpositus inactivation did not impair trajectory adaptation to reach over the obstacle. During inactivation of either interpositus nucleus, all measures of kinematic temporal and spatial variability increased with somewhat greater effects being produced by anterior interpositus inactivation. We discuss our results in relation to the hypothesis that anterior and posterior interpositus have different roles in trajectory control, related possibly to feed-forward use of cutaneous and proprioceptive inputs, respectively. The loss of adaptive reprogramming during anterior interpositus inactivation further suggests a role in motor learning. Comparison with results from our earlier motor cortical study shows that the distinctive impairments produced by inactivation of these two nuclei are similar to those produced by selective inactivation of different zones in the forelimb area of rostral motor cortex. Our findings are consistent with the hypothesis that there are separate functional output channels from the anterior and posterior interpositus nuclei to rostral motor cortex for distinct aspects of trajectory control and, from anterior interpositus alone, for trajectory adaptation.


Crash rates of convertible cars.
Convertible cars have existed since among the first automobiles, and the lack of substantial roof structure creates some safety concerns. Though crash tests have demonstrated that convertibles can resist excessive intrusion in front and side crashes and that strong A-pillars and roll bars can help maintain survival space in rollovers, little work has been done examining the real-world crash experience of these vehicles. The objective of this study was to compare the crash experience of recent convertibles with nonconvertible versions of the same cars using the most recent crash data. Crash and exposure data were obtained from the U.S. Department of Transportation and IHS Markit, respectively. Rates of driver deaths and police-reported crash involvements were compared for 1- to 5-year-old convertible cars and their nonconvertible versions during 2014-2018. Exposure measures included registered vehicle years (RVY) and vehicle miles traveled (VMT). These rates were compared using the standardized mortality ratio to account for possible differences in exposure distribution. Crash circumstances (e.g., point of impact, rollover, ejection) and behavioral outcomes (e.g., speeding, alcohol impairment, seat belt use) were compared for drivers killed in crashes. Convertibles had lower driver death rates and police-reported crash involvement rates on the basis of both RVY and VMT. However, the differences in driver death rates were not statistically significant. Driver deaths per 10 billion VMT were 11% lower for convertibles, and driver involvement in police-reported crashes per 10 million VMT was 6% lower. On average, convertibles were driven 1,595 fewer miles per year than the nonconvertible versions of these cars. Among fatally injured drivers, convertibles had slightly higher rates of ejection, and behavioral differences were minimal. The number of rollovers was small and their rate did not substantially differ between convertibles and their nonconvertible versions. Safety concerns associated with convertibles' retractable roof structures were not supported by the results of this study.


Interactions between Egr1 and AP1 factors in regulation of tyrosine hydroxylase transcription.
Several treatments which regulate tyrosine hydroxylase (TH) transcription, such as stress in vivo, or 12-O-tetradecanoylphorbol-13-acetate (TPA) in cell culture, induce both Egr1 and AP1 factors. Previously, we identified a functional Egr1 motif overlapping with Sp1 site in the rat TH promoter. Its response to Egr1 also required the presence of an AP1/Ebox motif. Here, we further examined the cross-talk between these sites. Insertion of 10- or 20-bp between the Sp1/Egr1 and AP1/Ebox elements, reduced the ability of Egr1 to upregulate luciferase reporter activity controlled by the proximal 272 nucleotides of the rat TH promoter in PC12 cells. Electrophoretic mobility shift assays with nuclear extracts from TPA treated cells were used to identify the composition of the factors which bound the AP1/Ebox motif and whether there is competition with factors which bind the Sp1/Egr1 motif. The complexes formed with labeled AP1/E box oligonucleotide were reduced or supershifted with antisera to Fos family, c-Fos, Fra-2, and Jun D. Excess Sp1/Egr1 oligonucleotide or anti Egr1 antisera did not compete. Fra-2 was a major component of the complex after 2-4 h TPA. Transfection of PC12 cells with Fra-2 induced reporter activity requiring the AP1, but not the Egr1 motif. However, when cotransfected with Fra-2, Egr1 expression plasmids elicited lower induction of luciferase activity than observed with Egr1 alone. Our results suggest that although it does not compete for binding to the promoter, Egr1 can modulate the regulation of TH transcription by AP1 factors.


Efficacy of a stress management program for patients with hepatocellular carcinoma receiving transcatheter arterial embolization.
Transcatheter arterial embolization (TAE), a common treatment for patients with unresectable hepatocellular carcinoma (HCC), can provoke severe physical discomfort and psychologic stress. The purpose of this study was to investigate the effect of a combination of health education, muscle relaxation, and back massage on reducing physical and psychologic stress in HCC patients receiving TAE. A quasi-experimental design was used. Forty patients with HCC (30 men and 10 women) with a mean age of 57 +/- 12 years were recruited and randomly assigned to the control or experimental group. The effectiveness of the stress management program was evaluated using a knowledge questionnaire, a worry inventory, a state-trait anxiety inventory, and a physical distress scale. After completing the stress management program, the experimental group had a greater mean increase in knowledge score than the control group (5.1 vs 0.8, p < 0.0001) and a greater mean decrease in worry score (-8.2 vs 1.1, p < 0.0001). The mean decrease in the anxiety score in the experimental group was also significantly greater than in the control group before TAE (-5.8 vs 3.2, p < 0.001) and 2, 4, 6, and 7 days after TAE (-8.2 vs 7.1, p < 0.001; -8.7 vs 3.2, p < 0.001; -9.8 vs -2.1, p < 0.05; -11 vs -0.9, p < 0.05). The patients in the experimental group had a smaller mean increase in physical distress score than the control group at 2, 4, 6, and 7 days after TAE (34.7 vs 50.2, 20.9 vs 29.6, 10.6 vs 18.2, 3.9 vs 11.2, all p < 0.05). This stress management program effectively reduces the stress of HCC patients undergoing TAE.


Madecassoside reduces ischemia-reperfusion injury on regional ischemia induced heart infarction in rat.
Madecassoside (MA), one of the principle terpenoids in Centella asiatica, has shown protect effect on isolated rat hearts and isolated cardiomyocytes against reperfusion injury in our previous studies. The aim of this study is to investigate if MA also protected against myocardial ischemia-reperfusion injury in vivo. The ischemia infarction model was established in rats. Left ventricular function was monitored during the ischemia-reperfusion period by a multi-channel recorder. After the ischemia-reperfusion process the infarcted areas were assessed. The levels of lactate dehydrogenase (LDH), creatinephosphokinase (CK), malondialdehyde (MDA), super-oxide dismutase (SOD) and C-reactive protein (CRP) in serum were determined. Cardiomyocytic apoptosis was measured by terminal-deoxynucleotidyl transferase mediated nick end labeling (TUNEL) staining. Pre-treatment with MA (50, 10 mg/kg) attenuated myocardial damage characteristic of decreasing infarct size, decreasing LDH and CK release. Activities of SOD were increased and MDA level increased obviously in control group whereas pretreatment with MA blunted the decrease of SOD activity, markedly reduced the level of MDA and the activity of CRP, and relieved myocardial cell apoptosis. These results suggest that MA has the protective effect on myocardial ischemia-reperfusion injury. This protection ability possibly due to its anti-lipid peroxidation, anti-inflammation and anti-apoptosis function and the enhancement of SOD activity.


Lung colonization by Aspergillus fumigatus is controlled by ZNF77.
Aspergillus fumigatus is a critical pathogen of humans. Exposure to A. fumigatus conidia occurs frequently but is normally cleared from the respiratory airways. In contrast, individuals with respiratory diseases are often highly colonized by fungi. Here, we use genome-edited epithelial cells to show that the genetic variant rs35699176 in ZNF77 causes loss of integrity of the bronchial epithelium and increases levels of extracellular matrix proteins. These changes promote A. fumigatus conidial adhesion, germination and growth. RNA-seq and LC/MS-MS analysis reveal rs35699176 upregulates vesicle trafficking leading to an increment of adhesion proteins. These changes make cells carrying rs35699176 more receptive to A. fumigatus in the early stages of infection. Moreover, patients with fungal asthma carrying rs35699176+/- have higher A. fumigatus loads in their respiratory airway. Our results indicate ZNF77 as a key controller of Aspergillus colonization and suggest its utility as a risk-marker for patient stratification.


Suppression of tobacco basic chitinase gene expression in response to colonization by the arbuscular mycorrhizal fungus Glomus intraradices.
A differentially displayed cDNA clone (MD17) was isolated from tobacco roots (nicotiana tabacum cv. Xanthi-nc) infected with the arbuscular mycorrhizal (AM) fungus Glomus intraradices. The isolated DNA fragment exhibited a reduced level of expression in response to AM establishment and 90% identity with the 3' noncoding sequence of two basic chitinases (EC 18.104.22.168) from N. tabacum. Northern (RNA) blots and Western blots (immunoblots), probed with tobacco basic chitinase gene-specific probe and polyclonal antibodies raised against the chitinase enzyme, yielded hybridization patterns similar to those of MD17. Moreover, the up-regulation of the 32-kDa basic chitinase gene expression in tobacco roots by (1,2,3)-thiadiazole-7-carbothioic acid S-methyl ester (BTH) was less effective in mycorrhizal roots than in nonmycorrhizal controls. Suppression of endogenous basic chitinase (32-kDa) expression was also observed in transgenic mycorrhizal plants that constitutively express the 34-kDa basic chitinase A isoform. When plants were grown with an increased phosphate supply, no suppression of the 32-kDa basic chitinase was obtained. These findings indicate that during the colonization and establishment of G. intraradices in tobacco roots, expression of the basic chitinase gene is down-regulated at the mRNA level.


Lymphoid cell proliferation in renal transplants: biologic and diagnostic implications.
It is unclear whether alloreaction develops in peripheral lymphoid organs and effector cells being recruited to the target organs, or the entire process of alloreaction can happen within the transplanted kidneys. Interstitial inflammatory cell (IIC) proliferation was evaluated by MIB-1 antigen immunostain and the rate expressed as positive cells/1000 cells. This rate was higher in acute cell-mediated rejection (ACR) (25.7, n = 14) compared with normal kidney (0.4, n = 8), acute tubular necrosis (1.2, n = 8), chronic allograft nephropathy (CAN, 2.4, n = 20), and native kidneys with diverse diseases (9.2, n = 63); but was comparable to that in CAN with significant IIC (20.6, n = 16). 10.1% and 8.3% of T lymphocytes underwent proliferation in ACR with or without CAN, whereas only rare B lymphocytes or macrophages showed this change (<1.2%), regardless of diagnostic categories. All biopsies diagnosed as ACR in conjunction with a high rate of MIB-1 + IIC and 9/12 biopsies with CAN and significant IIC in which ACR was diagnosed due to a high rate of MIB-1 + IIC, responded to anti-rejection therapy. Proliferation of IIC involves predominantly T lymphocytes. These observations provide support for the concept of in situ alloimmunization, and facilitate the diagnosis of ACR.


Measuring respiratory health in longitudinal social science surveys.
Objectively assessing respiratory health in longitudinal social science surveys would involve collecting pulmonary function measures on research participants, either in clinic settings or at home. These measures include indicators of volume (e.g., maximal amount of air blown in the first second of a forced exhalation) and airflow (maximal speed air is exhaled during a forced exhalation). Equipment options include office spirometry, portable spirometry, or home peak flow monitoring. Each option has different equipment and personnel costs. The types of research questions that could be answered using pulmonary function measures in longitudinal household surveys are quite broad, ranging from effects of socioeconomic status and race/ethnicity on respiratory health to social/environmental factors that contribute to respiratory health to the long-term social and economic consequences of respiratory health problems. Currently, such data are lacking. Given the potential payoffs in scientific knowledge, adding these measures to population-based surveys merits serious consideration.


The immunosuppressant rapamycin mimics a starvation-like signal distinct from amino acid and glucose deprivation.
RAFT1/FRAP/mTOR is a key regulator of cell growth and division and the mammalian target of rapamycin, an immunosuppressive and anticancer drug. Rapamycin deprivation and nutrient deprivation have similar effects on the activity of S6 kinase 1 (S6K1) and 4E-BP1, two downstream effectors of RAFT1, but the relationship between nutrient- and rapamycin-sensitive pathways is unknown. Using transcriptional profiling, we show that, in human BJAB B-lymphoma cells and murine CTLL-2 T lymphocytes, rapamycin treatment affects the expression of many genes involved in nutrient and protein metabolism. The rapamycin-induced transcriptional profile is distinct from those induced by glucose, glutamine, or leucine deprivation but is most similar to that induced by amino acid deprivation. In particular, rapamycin treatment and amino acid deprivation up-regulate genes involved in nutrient catabolism and energy production and down-regulate genes participating in lipid and nucleotide synthesis and in protein synthesis, turnover, and folding. Surprisingly, however, rapamycin had effects opposite from those of amino acid starvation on the expression of a large group of genes involved in the synthesis, transport, and use of amino acids. Supported by measurements of nutrient use, the data suggest that RAFT1 is an energy and nutrient sensor and that rapamycin mimics a signal generated by the starvation of amino acids but that the signal is unlikely to be the absence of amino acids themselves. These observations underscore the importance of metabolism in controlling lymphocyte proliferation and offer a novel explanation for immunosuppression by rapamycin.


[Association of DNA methyltransferase 3B gene polymorphism with early-onset schizophrenia].
To investigate the association of DNA methyltransferase 3B (DNMT3B) gene polymorphism with the development of early-onset schizophrenia. A single nucleotide polymorphism (rs6119954) of DNMT3B gene was genotyped in 279 early-onset schizophrenic patients and 395 healthy controls, using TaqMan SNP Genotyping Assays. To detect the interaction between the DNMT3B gene and environmental factors, the prenatal information of the patients was collected. Genotype distribution of the rs6119954 locus was significantly different between patients and controls (Chi-square = 12.27, P< 0.01). The frequency of the G allele of this locus was significantly higher in patients than in controls (Chi-square = 12.76, P< 0.01). The G allele was highly associated with an earlier age of onset (P= 0.026). No interaction between the DNMT3B gene and environmental factors was found. DNMT3B gene is associated with early-onset schizophrenia and rs6119954 may plays an important role in age of onset of schizophrenia.


The environmental limits to geographic range expansion in birds.
The environmental factors limiting species' ranges across broad geographic and taxonomic scales are central to questions regarding the geographic variation in biodiversity and impacts of environmental change. However, our understanding remains relatively limited owing to the small-scale, correlative nature of most previous analyses. In this study, we provide a global test of the environmental determinants of range limits in birds, using both stochastic and environmentally deterministic models of range expansion to simulate spatial patterns in the shape of species' distributions. We show that spatial variation in range shape can be well explained by the action of a few key climatic variables in limiting range expansion. Furthermore, we demonstrate that these patterns cannot be explained by random processes alone. In addition, our study also identifies important differences in the factors constraining range expansion between biogeographic realms and between widespread and rare species. Although temperature was the principal climatic determinant of range shape at a global scale, its effects were driven primarily by widespread species, with the ranges of rare taxa better predicted by gradients in precipitation or topography. These key results were robust with respect to the index used to quantify range shape. Our analysis helps resolve long running debates regarding the role of environmentally deterministic and stochastic processes in structuring biodiversity and extends our understanding of the factors limiting species' ranges and the response of broad scale biodiversity patterns to environmental change.


Characterization of performance on an automated visual recognition memory task in 7.5-month-old infants.
Infant looking behaviors measured during visual assessment paradigms may be more reliable predictors of long-term cognitive outcomes than standard measures such as the Bayley Scales of Infant Development typically used in environmental epidemiology. Infrared eye tracking technology offers an innovative approach to automate collection and processing of looking behavior data, making it possible to efficiently assess large numbers of infants. The goals of this study were to characterize infant looking behavior measures including side preference, fixation duration, and novelty preference using eye tracking and an automated version of an established visual recognition memory paradigm that includes both human faces and geometric figures as stimuli. An ancillary goal was to assess the feasibility of obtaining a precise measure of looking to the eye region of faces from the eye-tracking data. In this study, 309 7.5-month-old infants from a prospective birth cohort were assessed using a visual recognition memory (VRM) paradigm. Infrared eye tracking was used to record looking time as infants were shown nine blocks of trials with a pair of identical faces or shapes followed by two trials in which the familiar stimulus was paired with a novel one. Infants were assessed in one of four conditions: in conditions A and B, stimulus set 1 were the familiar stimuli and set 2 were novel; in conditions C and D, set 2 were familiar and set 1 novel. The novel stimuli were presented on the right first in conditions A and C and on the left first in conditions B and D. We observed a significant right side preference, which has not been reported before (57% of looking time spent looking at right side stimulus, p-value < 0.0001). Infants showed a preference for the novel stimuli similar to that published in prior studies (57-60% of looking time spent looking at the novel stimulus, p-value < 0.0001), as well as average fixation durations similar to previous studies. Infants also showed a strong preference for the eyes versus the rest of the face (p-value < 0.0001). Novelty preference was significantly higher when set 2 stimuli were novel (p-value < 0.0001), suggesting a preference among infants for set 2 stimuli compared to set 1 stimuli. The pattern of novelty preference across trials was significantly different between infants who saw the novel stimuli on the left first and those who saw them on the right first (p-value < 0.0001) but the overall mean novelty preference was not significantly different between these groups. There were also significant differences in average fixation duration and eyes preference measures across stimuli (p-values < 0.05). These findings show that VRM assessment can be automated for use in large-scale epidemiological studies using infrared eye tracking with looking behavior measure results similar to those obtained with standard non-automated methods, and that side and stimulus preferences are important modifiers of looking behavior that are critical to consider in this type of assessment.