Iron status biomarkers and C-reactive protein in children aged 19 to 72 months in Chile.
The Chilean Ministry of Health has combated iron deficiency through the delivery of fortified milk by the National Complementary Feeding Program (NCFP). To assess iron status and associations between biomarkers of iron status and serum C-reactive protein (CRP) in 218 beneficiaries of the NCFP aged 19 to 72 months in Santiago and Valparaiso, Chile. Blood was collected from a cross-sectional representative sample. Iron status (measured by hemoglobin, zinc protoporphyrin, and serum ferritin levels) and inflammation (according to CRP level) were determined. Serum CRP level was positively associated with serum ferritin and zinc protoporphyrin levels (r = 0.16 and r = 0.15; p = .0168 and p = .0290, respectively). Serum ferritin was higher among children with high CRP (> 10 mg/dL) than among those with low CRP (< or = 10 mg/dL) (p = .003). After adjustment for 10, 6, and 5 mg/L CRP, the prevalence of low serum ferritin changed from 56.4% without adjustment to 60.6%, 61.5%, and 42.7%, respectively, and the prevalence of high zinc protoporphyrin changed from 22.9% to 21.6%, 17.4%, and 17.9%, respectively. There were no differences between regions in biomarkers of iron status. There was no association between consumption of fortified milk and the prevalence of abnormal serum ferritin (< 15 microg/L) after adjustment for sex, age, and breastfeeding (OR, 1.00; 95% CI, 0.99 to 1.01; p = .288). After adjustment for 10 mg/L CRP, 5.5% were classified as having iron-deficiency anemia, 42.7% as having iron-deficiency erythropoiesis, 17.9% as having depleted iron stores, and 35.8% as having normal iron status. CONCLUSIONS. CRP level was positively associated with: serum ferritin and zinc protoporphyrin levels. Chilean children aged 19 to 72 months from Santiago and Valparaiso who were beneficiaries of the NCFP had a low prevalence of iron-deficiency anemia, a high prevalence of iron-deficiency erythropoiesis, and a moderate prevalence of depleted iron stores.


Stellate cell apoptosis by a soluble mediator from immortalized human hepatocytes.
Activated hepatic stellate cells (HSCs) are the major source of extracellular matrix in fibrosis and cirrhosis. In this study, we have investigated the role of hepatitis C virus (HCV) core protein induced immortalized human hepatocytes (IHH) on HSC growth. Preferential growth of IHH and apoptosis of activated human hepatic stellate cells (LX2) were observed upon coculture of these two cell types in a dual chamber or in the presence of conditioned medium (CM) from IHH. CM did not display a growth inhibitory role on other hepatic (Huh-7, HepG2, Hep3B and THLE) and non-hepatic (HeLa, MCF-7, and BHK) epithelial cells, indicating that the soluble mediator from IHH does not have a generalized effect on cell lines examined in our study. Further studies suggested that CM from IHH increased the expression of TRAIL receptors on LX2 cell surface, and induced apoptosis by a caspase dependent mechanism. Peptide mass fingerprinting of the purified soluble mediator from CM suggested that gelsolin fragments may play a role in apoptosis of LX2 cells. Taken together, our results suggested that a soluble mediator secreted from immortalized human hepatocytes plays an important role in hepatic stellate cell growth regulation.


The First Injection: Rates of Urinary Retention in Women With Urgency Incontinence Treated With Intravesical OnabotulinumtoxinA Injection.
The aim of the study was to describe the rate of symptomatic and asymptomatic urinary retention and catheterization in women undergoing initial intravesical onabotulinumtoxinA (BnTA) injection for urgency urinary incontinence (UUI). This retrospective chart review included women receiving initial 100 U of BnTA injection for UUI for 5 years. Straight-catheterized postvoid residuals (PVRs) were performed 2 weeks after the injection. Women without the sensation of incomplete bladder emptying, worsened urgency, inability to void, or suprapubic pain but with PVR of greater than 300 mL were characterized as having asymptomatic retention, whereas women with a PVR of greater than 150 and any of these symptoms were diagnosed with symptomatic retention. One hundred eighty-seven 187 patients received initial BnTA injection. The majority were postmenopausal (89%) and white (82%) with a mean age of 65 years and body mass index of 30 kg/m2. One-third of the cohort underwent baseline urodynamic studies. At 2 weeks after injection, 163 patients (87%) followed up, and 17 (10%) had either asymptomatic or symptomatic retention (2% and 8%, respectively). There were no differences in demographic or pretreatment urodynamic parameters in women with and without retention except that women who had previous anti-stress urinary incontinence procedures were more likely to experience retention (53% vs 18%, P = 0.002) despite similar baseline PVRs. We demonstrated that the rate of retention requiring catheterization after 100 U BnTA may be as high as 10% although only 5% develop PVRs for 300 mL and only 2% have asymptomatic retention for 300 mL.


Modeling vascular inflammation and atherogenicity after inhalation of ambient levels of ozone: exploratory lessons from transcriptomics.
Epidemiologic studies have linked inhalation of air pollutants such as ozone to cardiovascular mortality. Human exposure studies have shown that inhalation of ambient levels of ozone causes airway and systemic inflammation and an imbalance in sympathetic/parasympathetic tone. To explore molecular mechanisms through which ozone inhalation contributes to cardiovascular mortality, we compared transcriptomics data previously obtained from bronchoalveolar lavage (BAL) cells obtained from healthy subjects after inhalational exposure to ozone (200 ppb for 4 h) to those of various cell samples from 11 published studies of patients with atherosclerotic disease using the Nextbio genomic data platform. Overlapping gene ontologies that may be involved in the transition from pulmonary to systemic vascular inflammation after ozone inhalation were explored. Local and systemic enzymatic activity of an overlapping upregulated gene, matrix metalloproteinase-9 (MMP-9), was measured by zymography after ozone exposure. A set of differentially expressed genes involved in response to stimulus, stress, and wounding were in common between the ozone and most of the atherosclerosis studies. Many of these genes contribute to biological processes such as cholesterol metabolism dysfunction, increased monocyte adherence, endothelial cell lesions, and matrix remodeling, and to diseases such as heart failure, ischemia, and atherosclerotic occlusive disease. Inhalation of ozone increased MMP-9 enzymatic activity in both BAL fluid and serum. Comparison of transcriptomics between BAL cells after ozone exposure and various cell types from patients with atherosclerotic disease reveals commonly regulated processes and potential mechanisms by which ozone inhalation may contribute to progression of pre-existent atherosclerotic lesions.


Primary progressive apraxia of speech: clinical features and acoustic and neurologic correlates.
This study summarizes 2 illustrative cases of a neurodegenerative speech disorder, primary progressive apraxia of speech (AOS), as a vehicle for providing an overview of the disorder and an approach to describing and quantifying its perceptual features and some of its temporal acoustic attributes. Two individuals with primary progressive AOS underwent speech-language and neurologic evaluations on 2 occasions, ranging from 2.0 to 7.5 years postonset. Performance on several tests, tasks, and rating scales, as well as several acoustic measures, were compared over time within and between cases. Acoustic measures were compared with performance of control speakers. Both patients initially presented with AOS as the only or predominant sign of disease and without aphasia or dysarthria. The presenting features and temporal progression were captured in an AOS Rating Scale, an Articulation Error Score, and temporal acoustic measures of utterance duration, syllable rates per second, rates of speechlike alternating motion and sequential motion, and a pairwise variability index measure. AOS can be the predominant manifestation of neurodegenerative disease. Clinical ratings of its attributes and acoustic measures of some of its temporal characteristics can support its diagnosis and help quantify its salient characteristics and progression over time.


Impact of "Killer Immunoglobulin-Like Receptor /Ligand" Genotypes on Outcome following Surgery among Patients with Colorectal Cancer: Activating KIRs Are Associated with Long-Term Disease Free Survival.
Approximately 30% of patients with stage II/III colorectal cancer develop recurrence following surgery. How individual regulation of host mediated anti-tumor cytotoxicity is modified by the killer-cell immunoglobulin-like receptor (KIRs) genotype is essential for prediction of outcome. We analyzed the frequency of KIR and KIR ligand Human Leukocyte Antigen Class I genotypes, and their effects on recurrence and disease-free survival (DFS). Out of randomly selected 87 colorectal cancer patients who underwent R0 resection operations between 2005 and 2008, 29 patients whose cancers progressed within a median five-year follow-up period were compared with 58 patients with no recurrence within the same time period. Recurrent cases shared similar tumor stages with non-recurrent cases, but had different localizations. We used DNA isolated from pathological archival lymphoid and tumor tissues for KIR and KIR ligand (HLA-C, group C1, group C2, and HLA-A-Bw4) genotyping. Among cases with recurrence, KIR2DL1 (inhibitory KIR) and A-Bw4 (ligand for inhibitory KIR3DL1) were observed more frequently (p=0.017 and p=0.024); and KIR2DS2 and KIR2DS3 (both activating KIRs) were observed less frequently (p=0.005 and p=0.043). Similarly, in the non-recurrent group, inhibitory KIR-ligand combinations 2DL1-C2 and 2DL3-C1 were less frequent, while the activating combination 2DS2-C1 was more frequent. The lack of KIR2DL1, 2DL1-C2, and 2DL3-C1 improved disease-free survival (DFS) (100% vs. 62.3%, p=0.05; 93.8% vs. 60.0%, p=0.035; 73.6% vs. 55.9%, p=0.07). The presence of KIR2DS2, 2DS3, and 2DS2-C1 improved DFS (77.8% vs. 48.5%, p=0.01; 79.4% vs. 58.5%, p=0.003; 76.9% vs. 51.4%, p=0.023). KIR2DS3 reduced the risk of recurrence (HR=0.263, 95% CI = 0.080-0.863, p=0.028). The number of activating KIRs are correlated strongly with DFS, none/ one/ two KIR : 54/77/98 months (p=0.004). In conclusion the inheritance of increasing numbers of activating KIRs and lack of inhibitory KIRs, independent of tumor localization or stage, is associated with long-term DFS.


[Stroke mortality by residence and place of death: Paraná State, Brazil, 2007].
This study analyzed stroke mortality in Paraná State, Brazil, in 2007, according to residence and place of death, using the official Mortality Information System. For deaths in individuals 65 years and older and from 45 to 64 years of age, 17.6% and 29.8%, respectively, occurred at home. An association was observed between region of residence and less than three years of schooling (OR = 3.96) four to seven years of schooling (OR = 1.75), and race or skin color (non-white: OR = 1.39). Place of death was associated with age over 65 (OR= 1.69), less than three years of schooling (OR = 1.70), and residence in municipalities other than the respective headquarters of the Regional Health offices (OR = 1.33). The results suggested inequalities in access to hospital care at the time of death from stroke, according to region of residence and socioeconomic variables. The study supports use of the Mortality Information System as a data source to back public health policies and interventions. Further studies are needed to evaluate the factors that influence quality of patient care for stroke and to verify whether the same pattern is repeated in mortality from other causes.


Development of Endocyclic Control Elements for Peptide Macrocycles.
Synthetic methods that provide control over macrocycle conformation represent valuable tools for the discovery of bioactive molecules. Incorporation of heterocycles into cyclic peptides may offer a way to stabilize their solution conformations. Herein, we used N-(isocyanimino)triphenylphosphorane (Pinc) to install an oxadiazole ring and an endocyclic amine into peptide macrocycles. To elucidate the conformational effect of this constellation of functionalities, we performed synthesis, variable temperature NMR analysis, and NOE-based molecular dynamics simulation of a range of macrocycles in DMSO. As part of this study, we conducted experiments to compare macrocycle conformation in aqueous and DMSO solutions. The obtained solution structures suggest that the reduced amide bond/heterocycle (RAH) motif can stabilize macrocycle conformations in both water and DMSO, which addresses an enduring challenge in molecular design. The conformational effect of the RAH was used in an effort to mimic the biologically relevant secondary structure of MAdCAM-1. This resulted in the discovery of a novel α4β7 integrin antagonist.


Physiological role of electrogenic pumps in smooth muscle.
Rhythmic electrical activity in the small intestine of cat appears to be initiated by small pacemaker potentials originating in the longitudinal muscle layer. These depolarizing potentials (slow waves) have durations of from 2 to 5 sec and are generally less than 15 mV in amplitude. Frequency depends on the region of intestine and varies between 3 and 10 waves per min. Electrical activity of the intact muscle coat is the summation of this pacemaker activity with triggered activity and is of larger amplitude and more complicated waveform. Exposure to ouabain (10(-5) M) or K-free saline causes a rapid block of pacemaker slow-wave activity and depolarization to levels more positive than the slow-wave crest. Repolarization does not restore spontaneity. Amplitude and frequency are relatively insensitive to changes in extracellular Na and Cl concentrations, and conductance changes, if they occur during the slow-wave cycle, are too small to be detected in the double-sucrose gap. Changes in extracellular Ca have pronounced effects on frequency, being decreased in low Ca and increased in high Ca. Calcium channel blockers have the same effects as low Ca saline, implying that the changes in frequency are mediated through internal Ca levels. Experiments using chlorotetracycline fluorescence provided data that intracellular Ca2+ levels oscillate even under conditions where measurable contractile activity is absent. We have proposed that oscillations in the activity of the Na-K pump are responsible for the pacemaker slow waves and that internal Ca2+ is an important cycle-to-cycle controlling factor in the oscillation.


[Auditory functioning with cochlear implants of bimodal amplification].
The main function of a cochlear implant is to optimise the auditory and communicative abilities of individuals with hearing loss. The use of a hearing aid in the non-implanted ear further optimises binaural hearing, which leads to improved sound localization, speech discrimination in noise and quiet surroundings, and refined sound quality. The goal of the study was to determine the experiences of young adults with cochlear implants regarding bimodal amplification. Considering that all age groups are implanted with cochlear implants and that binaural amplification is beneficial to all age groups, it is important that information on young adults also be included. This study addressed bimodal amplification in terms of auditory, communicative, and cosmetic aspects, and handling and care of apparatus. A qualitative, descriptive research design was employed, and questionnaires were completed by five clients who are part of the Pretoria Cochlear Implant Program. Three of the participants took part in the focus group discussion in support of the results taken from the completed questionnaires. The results of the study indicate that the participants experience bimodal amplification positively, especially in terms of auditory and everyday communicative functioning. The participants largely perceived cosmetic aspects, handling and care of apparatus negatively. The outcomes of the study can be used by audiologists in order to adequately inform and counsel candidates regarding bimodal amplification. Potential barriers can also be addressed appropriately.


The versatility of propeller flaps for lower limb reconstruction in patients with peripheral arterial obstructive disease: initial experience.
In this study, we evaluate the feasibility of propeller flaps, for soft tissue reconstruction in patients with peripheral arterial obstructive disease (PAOD). Five patients (average age, 72 years old), presenting with necrosis of several foot units, were diagnosed with stage IV PAOD and diabetes mellitus. All underwent revascularization, followed by soft tissue reconstruction using propeller flaps. A total of 6 flaps were performed, with dimensions ranging from 4 x 7 cm to 8 x 31 cm. Perforator origin was the peroneal (5 flaps) or the tibial posterior artery (1 flap). Average operative time was 190 minutes. Overall patency was 83.3%; 1 case was complicated with total flap loss followed by below-knee amputation; 1 case developed partial necrosis necessitating skin grafting. All flaps developed mild postoperative edema, which resolved spontaneously. Propeller flaps prove to be useful tools for soft tissue reconstruction in patients with PAOD because of low operative morbidity, easy technique with fast learning curve, and low complications rate.


The insulin-like growth factor receptor I promotes motility and invasion of bladder cancer cells through Akt- and mitogen-activated protein kinase-dependent activation of paxillin.
The insulin-like growth factor receptor I (IGF-IR) plays an essential role in transformation by promoting cell growth and protecting cancer cells from apoptosis. Aberrant IGF-IR signaling is implicated in several types of tumors, including carcinomas of the lung, breast, prostate, pancreas, liver, and colon. However, the contribution of the IGF-IR to the development of the transformed phenotype in urothelial cells has not been clearly established. In this study we demonstrated that the IGF-IR is overexpressed in invasive bladder cancer tissues compared with nonmalignant controls. We have investigated the role of the IGF-IR in bladder cancer by using urothelial carcinoma-derived 5637 and T24 cells. Although activation of the IGF-IR did not appreciably affect their growth, it did promote migration and stimulate in vitro wound closure and invasion. These effects required the activation of the Akt and Mitogen-activated protein kinase (MAPK) pathways as well as IGF-I-induced Akt- and MAPK-dependent phosphorylation of paxillin, which relocated at dynamic focal adhesions and was necessary for promoting motility in bladder cancer cells. Our results provide the first evidence for a role of the IGF-IR in motility and invasion of bladder cancer cells and support the hypothesis that the IGF-IR may play a critical role in the establishment of the invasive phenotype in urothelial neoplasia. Thus, the IGF-IR may also serve as a novel biomarker for bladder cancer.


[Esophagoplasty with the colon in flap patients with burn strictures of the esophagus].
Results of coloesophagoplasty in 366 patients with long burn strictures of the esophagus are presented. The lesion was total in 74 (21.2%) of them. Inefficacy of bougieurage was indication for plastic surgery in 133 (36.3%) patients, complete obliteration of esophagus--in 53 (14.5%). Left half of the colon was used in 296 (80.9%) cases, transverse colon--in 48 (13.1%), right half of the colon--in 22 (6.0%) cases. Retrosternal shunting esophagoplasty was performed in 247 (67.5%) patients. Complications were in 84 (34.0%), necrosis of the transplant--in 4 (1.6%) patients. Extirpation of the esophagus with posteriormediastinoplasty was performed in 16 (4.4%) patients, in 14 of them--transhiatally. Complications occurred in 7 patients coloesophagopharyngoplasty was carried out in 52 (14.2%) patients with combined stricture of the esophagus and the pharynx, in 23 of them--simultaneously. Complications were in 28 (53.8%) patients. Intrapleural of coloesophagoplasty was performed in 51 patients. General hospital lethality was 3.3%. Long-term results were studied in 259 patients. Good results of esophagoplasty were seen in 135 (52.1%) patients, satisfactory--in 65 (25.1%), unsatisfactory--in 59 (22.8%) patients. Dysfunction of the artificial esophagus was seen in 22.0% patients, 12.7% patients underwent repeated reconstructive surgeries.


Analytical methodology and regulations for ochratoxin A.
Various methods exist to determine ochratoxin A. Currently they are often based on high performance liquid chromatography, but thin layer chromatography and enzyme-linked immunosorbent assay are also applied. Immunoaffinity column purification of extracts is an attractive approach to obtain clean extracts. Analytical quality assurance is essential to guarantee the integrity of analytical data and the use of reference materials plays an important role in this respect. (Proposed) regulations for ochratoxin A have been established in at least eight countries. The tolerance levels vary widely and the scientific basis for them appears to be weak.


An investigation of somatosensory profiles in work related upper limb disorders: a case-control observational study protocol.
Work related upper limb disorders constitute 45% of all occupational diseases and are a significant public health problem. A subgroup, non specific arm pain (NSAP), remains elusive in terms of understanding its pathophysiological mechanisms with its diagnosis based on the absence of specific clinical findings. One commonly proposed theory is that a neural tissue disorder is the primary dysfunction in NSAP and findings from previous studies lend some support to this theory. However, it is not clear if changes identified are simply a consequence of ongoing pain rather than due to specific neural changes. The presence of neuropathic pain has been investigated in several other musculoskeletal conditions but currently, there is no specific diagnostic tool or gold standard which permits an unequivocal diagnosis of neuropathic pain. The purpose of this study is to further describe the somatosensory profiles in patients with NSAP and to compare these profiles to a group of patients with MRI confirmed cervical radiculopathy who have been previously classified as having neuropathic pain. Three groups of participants will be investigated: Groups 1 and 2 will be office workers with either NSAP or cervical radiculopathy and Group 3 will be a control group of non office workers without upper limb pain. Participants will undergo a clinical assessment, pain questionnaires (LANSS, Short Form McGill, DASH and TSK) and quantitative sensory testing comprising thermal detection and pain thresholds, vibration thresholds and pressure pain thresholds. The spectrum of clinically suspected neuropathic pain ranges from more obvious conditions such as trigeminal neuralgia to those with vague signs of nerve disorder such as NSAP. A thorough description of the somatosensory profiles of NSAP patients and a comparison with a more defined group of patients with evidence of neuropathic pain will help in the understanding of underlying neurophysiology in NSAP and may influence future classification and intervention studies relating to this condition.


Osmotic regulation of root system architecture.
Although root system architecture is known to be highly plastic and strongly affected by environmental conditions, we have little understanding of the underlying mechanisms controlling root system development. Here we demonstrate that the formation of a lateral root from a lateral root primordium is repressed as water availability is reduced. This osmotic-responsive regulatory mechanism requires abscisic acid (ABA) and a newly identified gene, LRD2. Mutant analysis also revealed interactions of ABA and LRD2 with auxin signaling. Surprisingly, further examination revealed that both ABA and LRD2 control root system architecture even in the absence of osmotic stress. This suggests that the same molecules that mediate responses to environmental cues can also be regulators of intrinsic developmental programs in the root system.


Effects of tramadol hydrochloride on the thermal threshold in cats.
OBJECTIVE-To determine the thermal antinociceptive effect of oral administration of tramadol hydrochloride at doses between 0.5 and 4 mg/kg in cats. ANIMALS-6 healthy adult domestic shorthair cats. PROCEDURES-Baseline (before drug administration; time 0) thermal threshold was determined by applying a thermal probe to the thorax of each cat. Tramadol (0.5, 1, 2, 3, or 4 mg/kg) or a placebo was then administered orally in accordance with a Latin square design. Thermal threshold was determined by an observer who was unaware of treatment at various times until thermal threshold returned to baseline values or 6 hours had elapsed. Plasma tramadol and O-desmethyl-tramadol concentrations were measured prior to drug administration and at 1-hour intervals thereafter. Effect-concentration data were fitted to effect maximum models. RESULTS-Highest plasma tramadol and O-desmethyl-tramadol concentrations increased with increasing tramadol dose. Significant effects of dose and time on thermal threshold were detected. Thermal threshold was significantly higher than the baseline value at 80 and 120 minutes for the 0.5 mg/kg dose, at 80 and from 120 to 360 minutes for the 2 mg/kg dose, from 40 to 360 minutes for the 3 mg/kg dose, and from 60 to 360 minutes for the 4 mg/kg dose. CONCLUSIONS AND CLINICAL RELEVANCE-Tramadol induced thermal antinociception in cats. Doses of 2 to 4 mg/kg appeared necessary for induction of significant and sustained analgesic effects. Simulations predicted that 4 mg/kg every 6 hours would maintain analgesia close to the maximum effect of tramadol.


Escape of a plant virus from amplicon-mediated RNA silencing is associated with biotic or abiotic stress.
Strong RNA silencing was induced in plants transformed with an amplicon consisting of full-length cDNA of potato leafroll virus (PLRV) expressing green fluorescent protein (GFP), as shown by low levels of PLRV-GFP accumulation, lack of symptoms and accumulation of amplicon-specific short interfering RNAs (siRNAs). Inoculation of these plants with various viruses known to encode silencing suppressor proteins induced a striking synergistic effect leading to the enhanced accumulation of PLRV-GFP, suggesting that it had escaped from silencing. However, PLRV-GFP escape also occurred following inoculation with viruses that do not encode known silencing suppressors and treatment of silenced plants with biotic or abiotic stress agents. We propose that viruses can evade host RNA-silencing defences by a previously unrecognized mechanism that may be associated with a host response to some types of abiotic stress such as heat shock.


Adolescents' Communication with Parents, Other Adult Family Members and Teachers on Sexuality: Effects of School-Based Interventions in South Africa and Tanzania.
Cluster-randomized controlled trials were carried out to examine effects on sexual practices of school-based interventions among adolescents in three sites in sub-Saharan Africa. In this publication, effects on communication about sexuality with significant adults (including parents) and such communication as a mediator of other outcomes were examined. Belonging to the intervention group was significantly associated with fewer reported sexual debuts in Dar es Salaam only (OR 0.648). Effects on communication with adults about sexuality issues were stronger for Dar es Salaam than for the other sites. In Dar, increase in communication with adults proved to partially mediate associations between intervention and a number of social cognition outcomes. The hypothesized mediational effect of communication on sexual debut was not confirmed. Promoting intergenerational communication on sexuality issues is associated with several positive outcomes and therefore important. Future research should search for mediating factors influencing behavior beyond those examined in the present study.


False-positive tricyclic antidepressant drug screen results leading to the diagnosis of carbamazepine intoxication.
Ingestion of toxic substances is a common problem in pediatrics. When presented with the limited history of an unknown ingestion in a patient with altered mental status, a clinician depends on the physical examination and a toxic screen to determine the ingested substance(s). Some toxic screens yield false-positive or false-negative results that confound identification of ingested toxins. Three cases are presented in which carbamazepine ingestions were identified because of the false-positive tricyclic antidepressant serum toxic screen result in each case. Carbamazepine ingestion is one of the most common pediatric overdoses. Side effects include altered mental status, tachycardia, mydriasis, seizures, coma, and death. Several other substances also cause false-positive tricyclic antidepressant toxic screen results, including certain antipsychotic medications, antihistamines, and the muscle relaxant cyclobenzaprine. Specific tests and drugs causing false-positive results are presented in table form. More modern methods, specifically gas chromatographic-mass spectrometric, are more reliable in distinguishing these drugs. Knowledge of which substances commonly cause false-positive results on a given toxic screen can still lead the clinician to the correct diagnosis. tricyclic, carbamazepine, ingestion, intoxication, drug screen.


Gene encoding the catalytic subunit p110beta of human phosphatidylinositol 3-kinase: cloning, genomic structure, and screening for variants in patients with type 2 diabetes.
Phosphatidylinositol (PI) 3-kinase is a key signaling molecule in insulin-stimulated glucose transport. Therefore, we investigated the catalytic subunit p110beta, of human PI 3-kinase as a candidate gene for type 2 diabetes. Human p110beta gene was cloned from the placental genomic library. All 22 exons, intronic regions flanking the exons and 1.5 kb of the proximal/5' region of the p110beta gene, were screened for variants by single-strand conformation polymorphism analysis in 79 Finnish patients with type 2 diabetes . Allele frequencies of the variants were also determined in 77 nondiabetic control subjects. No variants were found in exons in diabetic patients. However, we identified two nucleotide polymorphisms in the proximal/5' region of the p110beta gene and a variation in the number of 2-bp repeat sequence (TA)n in intron 4. The allele frequencies did not differ between diabetic and control subjects. Our results may indicate that the catalytic subunit p110beta of PI 3-kinase plays such a fundamental role in the insulin-signaling pathway that structural variants are not likely to exist in that gene. The importance of the polymorphisms in the proximal/5' region of the p110beta gene for insulin signaling remains to be determined.


A longitudinal study of maternal cardiovascular function from preconception to the postpartum period.
Our objective was to investigate the extent of changes in maternal cardiovascular function, lipids and renal function during normal pregnancy from preconception to postpartum period. In this prospective study of 54 normal pregnancies, detailed hemodynamics were performed preconception, at 6, 23 and 33 weeks during pregnancy and 16 weeks postpartum. Although the greatest reduction of blood pressures (BPs) and augmentation index occurred in early pregnancy (Δbrachial systolic: 4 ± 7 mmHg, Δcentral systolic: 7 ± 7 mmHg; P < 0.001), the peripheral vascular resistance reached a nadir (Δ: 222 ± 215 dynes.s.cm; P < 0.001) by the second trimester. The greatest increase in cardiac output occurred by the second trimester (Δ: 0.6 ± 1 l/min, P < 0.001), whereas the heart rate increased maximally by the third trimester (Δ: 13 ± 11 bpm; P = 0.001). The unadjusted aortic pulse wave velocity decreased in the second trimester (P < 0.001), however, when adjusted for mean arterial pressure this was not significant (P = 0.06). BPs were lower (Δ brachial systolic: 5 ± 8 mmHg; P < 0.001) and augmentation index higher (Δ: 2.5 ± 7%; P = 0.01) postpartum than preconception. The cholesterol:high-density lipoprotein ratio, serum low density lipoprotein and serum creatinine all fell (P < 0.001) in the first trimester. We have shown that normal pregnancy, irrespective of parity, is associated with significant changes commencing very early in pregnancy, continuing throughout pregnancy, and some of these changes persisted postpartum. Therefore, first trimester or postpartum baselines will underestimate the true extent of pregnancy-related changes. Prospective studies of cardiovascular function from preconception to postpartum will provide more reliable estimates of the influence of cardiovascular maladaptation during pregnancy complications and their effect on longer term cardiovascular function.


Foot Salvage with Selective Arterialization of Pedal Venosomes.
Lower extremity salvage using microvascular flaps can be considered in high-functioning patients, but indications for salvage are often restricted by distal microvascular disease that may threaten the foot as a whole. Selective arterialization of specific pedal venosomes is a novel potential solution to this problem. A 48-year-old man with a chronic left plantar forefoot wound, osteomyelitis of the second and third metatarsal heads, and critical limb ischemia was evaluated for foot salvage. Despite a patent popliteal to posterior tibial bypass graft, residual ischemia was present in the first and second toes due to severe microvascular disease. This wound was reconstructed with a free partial medial rectus abdominis flow-through flap based on the medial branch of the deep inferior epigastric artery. The lateral branch was used as a flow-through vessel to arterialize the dorsal veins of the first webspace. Arterialization of the first webspace veins resulted in an immediate intraoperative improvement of the color of the first and second toes, along with creation of an arterial Doppler signal. The flap effectively covered the wound. The patient went on to heal completely and return to unrestricted ambulation. Transcutaneous oxygen measurement values of the dorsal foot improved from 35.8 mm Hg preoperatively to 48.3 mm Hg postoperatively. In patients with focal areas of critical ischemia, selective arterialization of the venous system may be an effective method of preventing tissue loss and improving wound healing. A flow-through flap can be used to accomplish this in tandem with wound reconstruction.


The effect of intramural and subserous uterine fibroids on implantation and clinical pregnancy rates in patients having intracytoplasmic sperm injection.
Our objective was to assess the effects of intramural and subserous fibroids on intracytoplasmic sperm injection (ICSI) in a retrospective case-control study of 108 women with uterine fibroids and 324 controls. The fibroids were located and measured by transvaginal ultrasound performed just before the ICSI cycle and all patients had normal endometrial scan. Seventy-three women had intramural and 35 women had subserous fibroids and the maximum diameter in any patient ranged from 0.5-10 cm. The number of fibroids in a patient ranged from 1-8. The first cycle outcome was compared with an age and body mass index matched 324 ICSI patients/cycles. All couples had male factor infertility. The ICSI cycles of patients with intramural and subserous fibroids were comparable in terms of the days of ovarian stimulation, the total dose of gonadotropin used, estradiol level on day of hCG administration, the number of metaphase II oocytes retrieved, fertilization and cleavage rates, number and quality of embryos developed and transferred. The implantation and clinical pregnancy rates were similar. We conclude that the presence of intramural and subserous fibroids does not adversely effect clinical pregnancy and implantation rates in patients undergoing ICSI.


Folding dynamics of a small protein at room temperature via simulated coherent two-dimensional infrared spectroscopy.
Understanding protein folding is of fundamental and practical importance in chemistry and biology. Despite the great success that has been made in tackling this problem, a detailed knowledge of how the elementary processes such as hydrogen-bond formation occur during protein folding has remained largely elusive. Using the combined power of molecular dynamics simulation with electrostatic polarization and coherent two-dimensional infrared spectroscopy, we are able to delineate the order of the hydrogen-bond formation event of a 17-residue peptide during its folding from an extended state to the native α-helix state. The folding is carried out by a single trajectory room-temperature molecular dynamics simulation that includes the polarization effect of hydrogen bonding, which is critical to the successful folding of the peptide. The onset and evolution of the isotope-labeled amide I vibration diagonal and cross peaks on the simulated 2DIR spectra allow us to build a structure-spectrum connection, and thus provide a microscopic picture of the helix folding process.


Sex differences in the delayed impact of acute stress on the amygdala.
There is accumulating evidence that stress triggers specific temporal patterns of morphological plasticity in the amygdala, a brain area that plays a pivotal role in the debilitating emotional symptoms of stress-related psychiatric disorders. Acute immobilization stress is known to cause a delayed increase in the density of dendritic spines on principal neurons in the basolateral amygdala (BLA) of rats. These neuronal changes are also accompanied by a delayed enhancement in anxiety-like behavior. However, these earlier studies used male rats, and the delayed behavioral and synaptic effects of acute stress on the BLA of female rats remain unexplored. Here, using whole-cell recordings in rat brain slices, we find that a single exposure to 2-h immobilization stress leads to an increase, 10 days later, in the frequency of miniature excitatory postsynaptic currents (mEPSCs) recorded from lateral amygdala (LA) principal neurons in male rats. Further, acute stress also causes a reduction in the frequency of miniature inhibitory postsynaptic currents (mIPSCs) in LA neurons 10 days after acute stress. In striking contrast, excitatory and inhibitory synaptic transmission in the LA of female rats does not exhibit any delayed change despite exposure to the same acute stress. Finally, we examined the functional impact of these contrasting synaptic changes at the behavioral level. Male rats exhibit a delayed increase in anxiety-like behavior on the elevated plus-maze 10 days after acute stress. However, the same stress does not lead to a delayed anxiogenic effect in female rats. Together, these results demonstrate that the delayed modulation of the balance of synaptic excitation and inhibition in the amygdala, as well as anxiety-like behavior, differ between males and females. These findings provide a framework, across biological scales, for analyzing how affective symptoms of stress disorders vary between the sexes.


Living on the edge: Conservation genetics of seven thermophilous plant species in a High Arctic archipelago.
Small, isolated, and/or peripheral populations are expected to harbour low levels of genetic variation and may therefore have reduced adaptability to environmental change, including climate warming. In the Arctic, global warming has already caused vegetation change across the region and is acting as a significant stressor on Arctic biodiversity. Many of the rare plants in the Arctic are relicts from early Holocene warm periods, but their ability to benefit from the current warming is dependent on the viability of their populations. We therefore examined Amplified Fragment Length Polymorphism (AFLP) data from regional red listed vascular plant species in the High Arctic archipelago of Svalbard and reference populations from the main distribution area of: 1) Botrychium lunaria, 2) Carex capillaris ssp. fuscidula, 3) Comastoma tenellum, 4) Kobresia simpliciuscula ssp. subholarctica, 5) Ranunculus wilanderi, 6) Sibbaldia procumbens and 7) Tofieldia pusilla In addition, we gathered population size data in Svalbard. The Svalbard populations had low genetic diversity and distinctiveness and few or no private markers compared to populations outside the archipelago. This is similar to observations in other rare species in Svalbard and the genetic depletion may be due to an initial founder effect and/or a genetic bottleneck caused by late Holocene cooling. There seems to be limited gene flow from other areas and the Svalbard populations should therefore be considered as demographically independent management units. Overall, these management units have small and/or few populations and are therefore prone to stochastic events which may further increase vulnerability to inbreeding depression, loss of genetic variation, and reduced evolutionary potential. Our results support theory predicting lower levels of genetic diversity in small, isolated and/or peripheral populations and may be of importance for management of other rare plant species in the Arctic.


Population Genetic Structure of the Giant Cactus Echinopsis terscheckii in Northwestern Argentina Is Shaped by Patterns of Vegetation Cover.
Species inhabiting drylands commonly depend on the surrounding vegetation for recruitment under stress, while competition may affect populations in moister environments. Our objective was to analyze how different climates and vegetation affect the fine-scale spatial genetic structure (SGS) of the columnar cactus Echinopsis terscheckii. At 4 sites, we estimated vegetation cover by digitized patches and the normalized difference vegetation index (NDVI). We mapped 30 individuals per population and collected tissue for isozyme electrophoresis using 15 putative loci. Spatial autocorrelation between all possible genotype pairs and the number of genetically homogeneous groups and families were calculated for each population. Greater cover (66%) and average NDVI values were detected in the most humid habitat that consisted of fewer, larger, and more dispersed vegetation patches. All populations were genetically diverse and showed significant SGS. Positive correlations were found between the distance at which maximum autocorrelation and kinship values were reached and vegetation area and patch size. Also higher NDVI values were associated with lower number of patches. Populations exposed to higher precipitation and vegetation cover consisted of sparse individuals that clustered at larger distances whereas vegetation patches in arid climates produced groups of closely related genotypes at small distances. These results support the stress-gradient genetic hypothesis. Under water stress, facilitation promotes establishment underneath patchy vegetation resulting in fine-scale family structure. In moister xerophilous forests, competition for resources, that is, light, results in sparse individuals and thus coarse-scale neighborhoods. This information can guide conservation and/or restoration efforts, such as the spatial scale to be considered in germplasm collection.


Intratumoral spatial heterogeneity of BTK kinomic activity dictates distinct therapeutic response within a single glioblastoma tumor.
Despite significant recent efforts applied toward the development of efficacious therapies for glioblastoma (GBM) through exploration of GBM's genome and transcriptome, curative therapeutic strategies remain highly elusive. As such, novel and effective therapeutics are urgently required. In this study, the authors sought to explore the kinomic landscape of GBM from a previously underutilized approach (i.e., spatial heterogeneity), followed by validation of Bruton's tyrosine kinase (BTK) targeting according to this stepwise kinomic-based novel approach. Twelve GBM tumor samples were obtained and characterized histopathologically from 2 patients with GBM. PamStation peptide-array analysis of these tissues was performed to measure the kinomic activity of each sample. The Ivy GBM database was then utilized to determine the intratumoral spatial localization of BTK activity by investigating the expression of BTK-related transcription factors (TFs) within tumors. Genetic inhibition of BTK family members through lentiviral short hairpin RNA (shRNA) knockdown was performed to determine their function in the core-like and edge-like GBM neurosphere models. Finally, the small-molecule inhibitor of BTK, ONO/GS-4059, which is currently under clinical investigation in nonbrain cancers, was applied for pharmacological inhibition of regionally specified newly established GBM edge and core neurosphere models. Kinomic investigation identified two major subclusters of GBM tissues from both patients exhibiting distinct profiles of kinase activity. Comparatively, in these spatially defined subgroups, BTK was the centric kinase differentially expressed. According to the Ivy GBM database, BTK-related TFs were highly expressed in the tumor core, but not in edge counterparts. Short hairpin RNA-mediated gene silencing of BTK in previously established edge- and core-like GBM neurospheres demonstrated increased apoptotic activity with predominance of the sub-G1 phase of core-like neurospheres compared to edge-like neurospheres. Lastly, pharmacological inhibition of BTK by ONO/GS-4059 resulted in growth inhibition of regionally derived GBM core cells and, to a lesser extent, their edge counterparts. This study identifies significant heterogeneity in kinase activity both within and across distinct GBM tumors. The study findings indicate that BTK activity is elevated in the classically therapy-resistant GBM tumor core. Given these findings, targeting GBM's resistant core through BTK may potentially provide therapeutic benefit for patients with GBM.


Regulator of G protein signaling protein 6 alleviates acute lung injury by inhibiting inflammation and promoting cell self-renewal in mice.
Acute respiratory distress syndrome (ARDS) is a disease with high mortality and morbidity. Regulator of G protein signaling protein 6 (RGS6), identified as a tumor suppressor gene, has received increasing attention owing to its close relationship with oxidative stress and inflammation. However, the association between ARDS and RGS6 has not been reported. Congruously regulated G protein-coupled receptor (GPCR)-related genes and differentially expressed genes (DEGs) in an acute lung injury (ALI) model were identified, and functional enrichment analysis was conducted. In an in vivo study, the effects of RGS6 knockout were studied in a mouse model of ALI induced by lipopolysaccharide (LPS). HE staining, ELISA, and immunohistochemistry were used to evaluate pathological changes and the degree of inflammation. In vitro, qRT‒PCR, immunofluorescence staining, and western blotting were used to determine the dynamic changes in RGS6 expression in cells. The RGS6 overexpression plasmid was constructed for transfection. qRT‒PCR was used to assess proinflammatory factors transcription. Western blotting and flow cytometry were used to evaluate apoptosis and reactive oxygen species (ROS) production. Organoid culture was used to assess the stemness and self-renewal capacity of alveolar epithelial type II cells (AEC2s). A total of 110 congruously regulated genes (61 congruously upregulated and 49 congruously downregulated genes) were identified among GPCR-related genes and DEGs in the ALI model. RGS6 was downregulated in vivo and in vitro in the ALI model. RGS6 was expressed in the cytoplasm and accumulated in the nucleus after LPS stimulation. Compared with the control group, we found higher mortality, more pronounced body weight changes, more serious pulmonary edema and pathological damage, and more neutrophil infiltration in the RGS6 knockout group upon LPS stimulation in vivo. Moreover, AEC2s loss was significantly increased upon RGS6 knockout. Organoid culture assays showed slower alveolar organoid formation, fewer alveolar organoids, and impaired development of new structures after passaging upon RGS6 knockout. In addition, RGS6 overexpression decreased ROS production as well as proinflammatory factor transcription in macrophages and decreased apoptosis in epithelial cells. RGS6 plays a protective role in ALI not only in early inflammatory responses but also in endogenous lung stem cell regeneration.


Do faculty and resident physicians discuss their medical errors?
Discussions about medical errors facilitate professional learning for physicians and may provide emotional support after an error, but little is known about physicians' attitudes and practices regarding error discussions with colleagues. Survey of faculty and resident physicians in generalist specialties in Midwest, Mid-Atlantic and Northeast regions of the US to investigate attitudes and practices regarding error discussions, likelihood of discussing hypothetical errors, experience role-modelling error discussions and demographic variables. Responses were received from 338 participants (response rate = 74%). In all, 73% of respondents indicated they usually discuss their mistakes with colleagues, 70% believed discussing mistakes strengthens professional relationships and 89% knew at least one colleague who would be a supportive listener. Motivations for error discussions included wanting to learn whether a colleague would have made the same decision (91%), wanting colleagues to learn from the mistake (80%) and wanting to receive support (79%). Given hypothetical scenarios, most respondents indicated they would likely discuss an error resulting in no harm (77%), minor harm (87%) or major harm (94%). Fifty-seven percent of physicians had tried to serve as a role model by discussing an error and role-modelling was more likely among those who had previously observed an error discussion (OR 4.17, CI 2.34 to 7.42). Most generalist physicians in teaching hospitals report that they usually discuss their errors with colleagues, and more than half have tried to role-model discussions. However, a significant number of these physicians report that they do not usually discuss their errors and some do not know colleagues who would be supportive listeners.


Funnel plots and choropleth maps in cancer risk communication: a comparison of tools for disseminating population-based incidence data to stakeholders.
Population-based cancer registries provide epidemiological cancer information, but the indicators are often too complex to be interpreted by local authorities and communities, due to numeracy and literacy limitations. The aim of this paper is to compare the commonly used visual formats to funnel plots to enable local public health authorities and communities to access valid and understandable cancer incidence data obtained at the municipal level. A funnel plot representation of standardised incidence ratio (SIR) was generated for the 82 municipalities of the Palermo Province with the 2003-2011 data from the Palermo Province Cancer Registry (Sicily, Italy). The properties of the funnel plot and choropleth map methodologies were compared within the context of disseminating epidemiological data to stakeholders. The SIRs of all the municipalities remained within the control limits, except for Palermo city area (SIR=1.12), which was sited outside the upper control limit line of 99.8%. The Palermo Province SIRs funnel plot representation was congruent with the choropleth map generated from the same data, but the former resulted more informative as shown by the comparisons of the weaknesses and strengths of the 2 visual formats. Funnel plot should be used as a complementary valuable tool to communicate epidemiological data of cancer registries to communities and local authorities, visually conveying an efficient and simple way to interpret cancer incidence data.


Oxytocin and sexual behaviour in the male rat and rabbit.
In male New Zealand white rabbits, it was shown that oxytocin but not vasopressin concentrations in plasma were markedly raised after ejaculation. In male Wistar rats, oxytocin infused into the internal carotid artery reduced the number of intromissions made before ejaculation but had no other significant effect. Infusion of oxytocin into the third ventricle increased the latencies to the first mount and intromission and lengthened post-ejaculatory refractory periods. It is suggested that oxytocin released into the periphery during coitus, while not essentially involved in ejaculation, may exert effects on the genital periphery. Behavioural effects of centrally administered oxytocin suggest that it may play a role in the neural mechanisms underlying post-ejaculatory refractoriness.


Recursion formula for reflectance and the enhanced effect on the light group velocity control of the stratified and phase-shifted volume index gratings.
We derived a recursion formula for the reflectance of the stratified and phase-shifted volume index gratings. The characteristics of the reflectance spectra of the stratified and phase-shifted volume index gratings were studied based on the recursion formula. It is shown that narrow bandwidth transparency peaks appear within the stop-band of the reflectance spectrum of the volume index gratings due to the intervention of the homogeneous buffer layers that induce the phase-shifts between neighboring volume index gratings. The spectral positions of the transparency peaks can be shifted within the stop-band by controlling the phase-shift, i.e., the buffer layer thickness. The described properties may find applications in addressable band-pass filter, switching, wavelength division multiplexing, and de-multiplexing. The dispersion near the transparency peaks of the stratified and phase-shifted volume index grating is found to be sharply enhanced as compared to the uniform volume index gratings. Significantly enhanced control on the group velocity of light by several orders of magnitude while keeping high transmittance is demonstrated in the stratified and phase-shifted volume index grating.


False-positive radio-iodinated metaiodobenzylguanidine (123I-MIBG) accumulation in a mast cell-infiltrated infantile haemangioma.
Radio-iodinated metaiodobenzylguanidine ((123)I-MIBG) is used for the detection and staging of neuroblastoma, pheochromcytoma and other neuroendocrine tumours in diagnostic nuclear medicine. A specific uptake and storage mechanism provides the basis for imaging with (123)I-MIBG. Nevertheless, cases of false-positive (123)I-MIBG scintigraphy with accumulation in non-chromaffin tumours have been described. Here, we present a case of a false-positive (123)I-MIBG scan in a case of a mast-cell infiltrated infantile haemangioma and discuss the possible uptake mechanism.


Prostate segmentation algorithm using dyadic wavelet transform and discrete dynamic contour.
Knowing the location and the volume of the prostate is important for ultrasound-guided prostate brachytherapy, a commonly used prostate cancer treatment method. The prostate boundary must be segmented before a dose plan can be obtained. However, manual segmentation is arduous and time consuming. This paper introduces a semi-automatic segmentation algorithm based on the dyadic wavelet transform (DWT) and the discrete dynamic contour (DDC). A spline interpolation method is used to determine the initial contour based on four user-defined initial points. The DDC model then refines the initial contour based on the approximate coefficients and the wavelet coefficients generated using the DWT. The DDC model is executed under two settings. The coefficients used in these two settings are derived using smoothing functions with different sizes. A selection rule is used to choose the best contour based on the contours produced in these two settings. The accuracy of the final contour produced by the proposed algorithm is evaluated by comparing it with the manual contour outlined by an expert observer. A total of 114 2D TRUS images taken for six different patients scheduled for brachytherapy were segmented using the proposed algorithm. The average difference between the contour segmented using the proposed algorithm and the manually outlined contour is less than 3 pixels.


Subserosal injection of hyaluronic acid may prevent perforation after endoscopic resection.
To investigate the protective effect of subserosal injection of hyaluronic acid (HA) after endoscopic resection (ER) using ex vivo and in vivo studies. As the first examination, technical application of subserosal injection was tested 10 times using resected porcine stomachs. As the second examination, ER was applied to make six mucosal defects per stomach in three live minipigs and thermal damage was given on the proper muscle layer by using hemostatic forceps. Following the thermocoagulation, 1 mL of normal saline and HA, respectively, was injected targeting the subserosal layer in two mucosal defects each and the rest kept no injection as the control. The minipigs were recovered from the anesthesia and kept fasting until euthanasia which was carried out around 24 h after the procedures. Ex vivo study revealed that complete and partial subserosal injection was possible two (20%) and four (40%) times, respectively. In vivo study revealed that no postoperative perforation occurred at any point of the thermocoagulation. Apparent retention of hyaluronic acid was identified at only two (33%) points where HA was injected. This study failed to show preventative effects of subserosal injection of HA on postoperative perforation due to technical faults. However, this concept has a possibility to change strategy of ER with further technical innovation.


Short term histopathological effects of GaAlAs laser on experimentally induced TMJ osteoarthritis in rabbits.
The aim of this study was to evaluate the biostimulation (BS) effect of the gallium-aluminum-arsenide (GaAlAs) diode laser by histopathology with an experimental osteoarthritis (OA) model in the temporomandibular joints (TMJ) of rabbits, in the early period. GaAlAs diode laser is used for pain reduction in TMJ disorders. Twenty-four adult male New Zealand white rabbits were randomly divided into three equal groups: Control Group (CG), Study Group 1 (SG-1), and Study Group 2 (SG-2). Mono-iodoacetate (MIA) was administered to the right TMJs of all rabbits. The rabbits did not undergo any treatment for four weeks to allow the development of osteoarthritis. In SG-1, laser BS was applied to the rabbits at 940 nm, 5 W, and 15 J/cm2 in continuous wave mode at 48-hour intervals for 14 sessions; and in SG-2, laser BS was applied with the same parameters at 24-hour intervals for 28 sessions. Laser BS was not applied to the rabbits in CG. All rabbits were sacrificed simultaneously. The TMJ cartilage, osteochondral junction, chondrocyte appearance, and subchondral ossification were evaluated histopathologically. There was no statistically significant difference between the groups in terms of cartilage, osteochondral junction, chondrocyte appearance, and subchondral ossification values (p > 0.05). The laser BS protocol used in the study had no positive histopathological effects on TMJ OA in the early period.


Expansion and Retroviral Transduction of Primary Murine T Cells for CAR T-Cell Therapy.
The development of chimeric antigen receptor (CAR) T cells has been a revolutionary technology for the treatment of relapsed and refractory leukemias and lymphomas. The synthetic CAR molecule redirects T cell function toward tumor surface-expressed antigens through a single-chain variable fragment (scFv) fused to CD3z and intracellular costimulatory domains. Here, we describe a protocol for the generation of CAR T cells using primary mouse T cells and a gammaretroviral vector encoding a CAR transgene. This protocol outlines several transduction and expansion methods based on the use of two transduction enhancers, RetroNectin® and Vectofusin®-1, and cell culture systems such as conventional plates or G-Rex® devices.


[Function of an inhibitor of cyclin-dependent kinase p27/Kip in cells transformed by E1A + E1B19 kDa + E1A + cHa-Ras, differing in their ability to realize a G1-block during serum starvation].
We studied the capability of E1A + cHa-ras and E1A + E1B19kDa transformants to undergo the G1/S arrest of the cell cycle following depletion of serum growth factors. It has been shown that serum starvation induced the G1/S arrest both in normal rat embryo fibroblasts (REF) and in E1A + E1B19kDa transformants, whereas E1A + cHa-ras transformed cells lost this feature. To analyse the mechanisms underlying these differences, we studied the expression of p27/KIP, its intracellular distribution and association with E1A oncoproducts. The content of the p27/KIP inhibitor of cyclin-dependent kinases was found to change a little upon transformation by two complementary oncogene pairs. However, serum starvation for 24 h led to a significant increase in the content of p27/KIP in E1A + E1B19kDa transformants, while E1A + cHa-ras cells accumulated p27/KIP less markedly. According to the immunofluorescence study, the p27/KIP inhibitor is located in the nucleus of both normal and transformed cells. Moreover, serum starvation did not lead to its inhibition due to redistribution to the cytoplasm in both cell lines. Also, we were unable to detect association of p27/KIP with E1A oncoproducts in immunoprecipitated complexes. The obtained data indicate that, in contrast to E1A + cHa-ras transformants, in E1A + E1B19kDa cells the p27/KIP inhibitor is functional and it is capable of inducing the G1/S block after serum starvation.


A novel copper-binding protein with characteristics of a metallothionein from a clinical isolate of Candida albicans.
It is known that clinical isolates of Candida albicans exhibit a high level of resistance to copper salts, although the molecular basis of this resistance is not clear. To investigate this, a novel copper-binding protein was purified from a clinical isolate of C. albicans. The protein was extracted from yeast cells after an induction period of 10 h in a copper-containing suspension medium. It was further purified by size-exclusion chromatography, ultrafiltration and reverse-phase HPLC. All protein fractions were analysed for their protein and copper contents. The copper/protein ratio increased steadily throughout the purification process; the most highly purified fraction showed a 210-fold increase compared to the whole-cell extract, with a recovery of 0.03%. The molecular mass of the protein was 10,000 Da and a reconstitution study using the purified apoprotein suggested that the equivalent extent of Cu(I) binding was approximately 14 mol eq. The amino-terminal segment of the copper-binding protein revealed three Cys-Xaa-Cys motifs, which is typical of a metallothionein (MT), and showed significant homology with mammalian MTs with respect to the positions of the cysteine residues. This is the first report of the isolation of a copper-binding protein from C. albicans.


Tuberculosis vaccines: past, present and future.
The current vaccine against tuberculosis protects against severe forms of the disease in children but confers variable effectiveness against pulmonary disease. With tuberculosis eradication on the horizon new vaccines with better protection than Mycobacterium bovis bacillus Calmette-Guérin (BCG) are needed. This review will outline the most promising tuberculosis vaccine candidates from selected publications. The enormous effort of the scientific community in the last 10 years has generated hundreds of tuberculosis vaccine candidates. These include sub-unit vaccines and live vaccines such as recombinant BCG and other attenuated live vaccines. Some of these are being included for the first time in phase I clinical trials. For more than 80 years now no new tuberculosis vaccine has successfully been developed. There is now renewed optimism that vaccines superior to BCG can be developed in the coming years. The goal is to obtain a new generation of vaccines effective against more transmissible forms of tuberculosis. As a first step, good candidate vaccines able to boost BCG and improve BCG protection could be a reality in the near future. Tuberculosis vaccine candidates, able to replace the currently used BCG and make the eradication of tuberculosis feasible, can be expected in the mid-term, and live vaccines are reliable and promising candidates.


Pollinators differ in their contribution to the male fitness of a self-incompatible composite.
Reproductive fitness in plants is often determined by the quantity and quality of pollen transferred by pollinators. However, many fitness studies measure only female fitness or rely on proxies for male fitness. Here we assessed how five bee taxon groups affect male fitness in a prairie plant by quantifying pollen removal, visitation, and siring success using paternity assignments and a unique pollinator visitation experiment. In Echinacea angustifolia, we measured per-visit pollen removal for each pollinator taxon and estimated the number of pollen grains needed for successful ovule fertilization. Additionally, we directly measured pollinator influence on siring by allowing only one bee taxon to visit each pollen-donor plant, while open-pollinated plants acted as unrestricted pollen recipients. We genotyped the resulting offspring, assigned paternity, and used aster statistical models to quantify siring success. Siring success of pollen-donor plants differed among the five pollinator groups. Nongrooming male bees were associated with increased siring success. Bees from all taxa removed most of the flowering head's pollen in one visit. However, coneflower-specialist bee Andrena helianthiformis removed the most pollen per visit. Female fitness and proxy measures of male fitness, such as pollinator visitation and pollen removal, did not align with our direct quantifications of male fitness. Our results illustrate the need for more studies to directly quantify male fitness, and we caution against using male fitness proxy measures. In addition, conservation efforts that preserve a diverse pollinator community can benefit plants in fragmented landscapes.


Smooth Pursuit and Saccades after Sport-Related Concussion.
Smooth pursuit eye movements (SPEMs) and saccadic eye movements are both commonly impaired following sport-related concussion (SRC). Typical oculomotor assessments measure individual eye movements in a series of restrictive tests designed to isolate features such as response times. These measures lack ecological validity for athletes because athletes are adept at simple tasks designed for the general population. Yet, because eye movement metrics are sensitive and well-characterized neuroanatomically, it would be valuable to test whether athletes exhibit abnormal eye movements with more challenging tasks. To address this gap in knowledge, we collected eye-tracking data during a sport-like task to gain insight on gaze behavior during active self-motion. SPEMs and saccadic eye movements were recorded during a sport-like visual task within 24-48 h following SRC. Thirty-six Division I student-athletes were divided into SRC and control (CON) groups. All participants completed two blocks of the Wii Fit© soccer heading game (WF) while wearing a monocular infrared eye tracker. Eye movement classification systems quantified saccadic amplitude (SA), velocity (SV), and count (SC); as well as SPEM velocity (SPV) and amplitude (SPA). Separate Mann-Whitney U tests evaluated SPA and SC and found no significant effects (SPA, p = 0.11; SC, p = 0.10). A multi-variate analysis of variance (MANOVA) for remaining variables revealed SPV was significantly greater in CON (p < 0.05), but the SRC group had greater SA and SV (p < 0.05). These findings suggest that during a sport-like task, to maintain foveation SRC subjects used larger amplitude, faster saccades, but exhibited slower SPEMs. Measuring oculomotor function during ecologically valid, sport-like tasks may serve as a concussion biomarker and provide insights into eye movement control after SRC.


[Studies on the iodide metabolism and the expression of thyroglobulin and thyroid peroxidase mRNA in the thyroid of BB/W rats].
BioBreeding/Worcester (BB/W) rats develop insulin dependent diabetes mellitus (IDDM) and lymphocytic thyroiditis (LT) spontaneously. Our previous studies have shown that BB/W (Saitama-Tokyo colony) rats develop LT at about 10 weeks of age. Their serum TSH values increase as LT extends, although their serum thyroid hormone levels remain normal. This indicates that BB/W rats suffer from subclinical hypothyroidism. To investigate whether BB/W rats have a defect in iodide metabolism, the thyroidal radioactive iodine uptake (RAIU) in BB/W rats was examined. Thyroidal RAIU at 3hr in both 8 and 16 week-old BB/W rats was significantly higher than that in age-matched normal Wistar rats. On the other hand, BB/W rats had significantly lower 48hr thyroidal RAIU than normal Wistar rats. This suggests that BB/W rats appear to have some defects in iodide metabolism, especially in iodide organification even before the development of LT. The expression of thyroid peroxidase (TPO) and thyroglobulin (Tg) mRNA in BB/W and Wistar rats was then examined using the Northern blot analysis. The expression of both TPO and Tg mRNA was greatly decreased in BB/W rats compared with that in Wistar rats despite the high serum TSH levels in BB/W rats. This indicates that BB/W rats may have pretranslational defects in TPO and Tg synthesis, resulting in the impaired thyroid hormone synthesis. In the present study, it has been demonstrated that BB/W rats appear to have a defect(s) in iodide metabolism possibly due to some abnormalities in TPO and Tg synthesis.


[Characteristics of isoenzymes of glucose-6-phosphate dehydrogenase in estrogen-dependent and estrogen-independent mammary tumors of experimental animals].
Estrogen-dependent and estrogen-independent mammary tumours were produced in experimental animals. Four isozymes of glucose 6-phosphate dehydrogenase were identified in each tumour tupe. It was found that ovariectomy and estradiol administration change selectively the activity of glucose 6-phosphate dehydrogenase I in estrogen-dependent tumours, while no effect on the enzyme or any of its isozymes was found in estrogen- in dependent tumours. Other hormones (prolactin, insulin) induce the activity of glucose 6-phosphate dehydrogenase III rather than that of glucose 6-phosphate dehydrogenase I in normal mammary tissue. The idea of existence of a fine regulation of the enzyme activity through selective activation or repression of individual isozymes is confirmed.


Stunting in infancy is associated with atypical activation of working memory and attention networks.
Stunting is associated with poor long-term cognitive, academic and economic outcomes, yet the mechanisms through which stunting impacts cognition in early development remain unknown. In a first-ever neuroimaging study conducted on infants from rural India, we demonstrate that stunting impacts a critical, early-developing cognitive system-visual working memory. Stunted infants showed poor visual working memory performance and were easily distractible. Poor performance was associated with reduced engagement of the left anterior intraparietal sulcus, a region involved in visual working memory maintenance and greater suppression in the right temporoparietal junction, a region involved in attentional shifting. When assessed one year later, stunted infants had lower problem-solving scores, while infants of normal height with greater left anterior intraparietal sulcus activation showed higher problem-solving scores. Finally, short-for-age infants with poor physical growth indices but good visual working memory performance showed more positive outcomes suggesting that intervention efforts should focus on improving working memory and reducing distractibility in infancy.


Microsatellite analysis reveals high genetic diversity but low genetic structure in Ethiopian indigenous cattle populations.
Ethiopian cattle are under threat from uncontrolled mating practices and are at high risk of becoming genetically homogeneous. Therefore, to evaluate genetic diversity, population structure and degree of admixture, 30 microsatellite markers were genotyped using 351 DNA samples from 10 Ethiopian cattle populations and the Holstein breed. The mean number of alleles per cattle population ranged from 6.93 +/- 2.12 in Sheko to 7.50 +/- 2.35 in Adwa. The mean observed and expected heterozygosities were 0.674 +/- 0.015 and 0.726 +/- 0.019 respectively. Ethiopian cattle populations have maintained a high level of within-population genetic differentiation (98.7%), the remainder being accounted for by differentiation among populations (1.3%). A highly significant deficiency in heterozygotes was detected within populations (F(IS) = 0.071; P < 0.001) and total inbreeding (F(IT) = 0.083; P < 0.001). The study populations were highly admixed but distinct from pure Bos taurus and Bos indicus breeds. The various levels of admixture and high genetic diversity make Ethiopian cattle populations suitable for future genetic improvement and utilization under a wide range of agro-ecologies in Ethiopia.


A dose-response evaluation of rumen-protected niacin in thermoneutral or heat-stressed lactating Holstein cows.
Twenty-four multiparous high-producing dairy cows (40.0±1.4kg/d) were used in a factorial design to evaluate effects of 2 environments [thermoneutral (TN) and heat stress (HS)] and a dose range of dietary rumen-protected niacin (RPN; 0, 4, 8, or 12g/d) on body temperature, sweating rate, feed intake, water intake, production parameters, and blood niacin concentrations. Temperature-humidity index values during TN never exceeded 68 (stress threshold), whereas temperature-humidity index values during HS were above 68 for 24h/d. The HS environment increased hair coat and skin, rectal, and vaginal temperatures; respiration rate; skin and hair coat evaporative heat loss; and water intake and decreased DMI (3.5kg/d), milk yield (4.1kg/d), 4% fat-corrected milk (2.7kg/d), and milk protein yield (181.7g/d). Sweating rate increased during HS (12.7g/m(2) per h) compared with TN, but this increase was only 10% of that reported in summer-acclimated cattle. Niacin supplementation did not affect sweating rate, dry-matter intake, or milk yield in either environment. Rumen-protected niacin increased plasma and milk niacin concentrations in a linear manner. Heat stress reduced niacin concentration in whole blood (7.86 vs. 6.89μg/mL) but not in milk. Reduced blood niacin concentration was partially corrected by dietary RPN. An interaction existed between dietary RPN and environment; dietary RPN linearly increased water intake in both environments, but the increase was greater during HS conditions. Increasing dietary RPN did not influence skin temperatures. During TN, supplementing 12g/d of RPN increased hair coat (unshaved skin; 30.3 vs. 31.3°C at 1600h) but not shaved skin (32.8 vs. 32.9°C at 1600h) temperature when compared with 0g/d at all time points, whereas the maximum temperature (18°C) of the room was lower than skin temperature. These data suggest that dietary RPN increased water intake during both TN and HS and hair coat temperature during TN; however, core body temperature was unaffected. Thus, encapsulated niacin did not improve thermotolerance of winter-acclimated lactating dairy cows exposed to moderate thermal stress in Arizona.


Naturally processed tissue- and differentiation stage-specific autologous peptides bound by HLA class I and II molecules of chronic myeloid leukemia blasts.
Structural analysis of naturally processed peptides bound to the HLA class I and class II molecules of chronic myeloid leukemia (CML) blast cells was performed to characterize the antigen processing and autoantigen repertoire in this hematopoietic malignancy. Self-peptides derived from the carboxy-terminal end of the breakpoint cluster region (bcr) protein, as well as several differentiation stage- and tissue-specific self-antigens characteristic of early stages of myeloid differentiation, such as c-fes, c-pim, granulocyte-macrophage colony-stimulating factor receptor alpha chain, proteinase 3, and cathepsin G, were identified. A common characteristic of several of the high copy-number self-peptides identified in this study is the participation of their parent proteins in signal transduction or myeloid effector function. Because bcr-abl junctional peptides bind to a limited number of major histocompatibility complex (MHC) class I alleles, an effective peptide-based immunotherapy strategy for CML requires identification of further tumor-associated or tissue-specific peptide antigens binding to common MHC alleles such as HLA-A2. The differentiation stage- and tissue-specific MHC-bound peptides found in this study, as well as the naturally processed proteins from which they are derived, may represent autoantigens towards which T-cell responses may potentially be developed for immunotherapy of hematopoietic malignancies such as CML.


Safety and efficacy of retroperitoneal sutureless zero ischemia laparoscopic partial nephrectomy for low nephrometry score masses.
To evaluate oncological feasibility and oncological and functional results of retroperitoneal sutureless zero ischemia laparoscopic partial nephrectomy (LPN). Patients with posterior renal masses with low nephrometry score (RENAL ≤ 7) treated who underwent retroperitoneal sutureless zero ischemia.in a single center from January 2016 to November 2017. Clinical, surgical and pathological data were prospectively collected. Complications were reported according to the modified Clavien classification. Retroperitoneal sutureless zero ischemia laparoscopic partial nephrectomy was performed on 15 patients. The indication for nephron-sparing surgery was elective in 11 (73%) patients and imperative in 4 (27%). Median RENAL score was 5 (IQR: 5-7), median tumor diameter 25 mm (IQR: 20-35). In 11 cases, the tumor was located polar (85%), and in 2 cases hilar (15%). There were no intraoperative complications. No cases were converted to radical nephrectomy, and in no case parenchyma suture was necessary. Median operative time was 90 min (IQR:40-150), in no case clamping of the renal artery was necessary, median hospital stay was 4 days, median estimated blood loss (EBL) was 310 (180-500) ml. Pathological analysis showed renal cell carcinoma in 11 patients (85%), 9 (60%) staged T1a and 2 (13%) T1b. In 4 (27%) an oncocytoma was found. There were no positive surgical margins. One patient developed a major postoperative complication (postoperative renal bleeding requiring super-selective embolization). Trifecta rate was 93%. Sutureless retroperitoneal zero ischemia LPN for the treatment of low-complexity posterior renal masses showed to be safe and feasible. Longer follow-up and higher numbers of patients are, however, warranted to draw definitive conclusions on functional outcomes.


A first evaluation of the usefulness of Kudzu starch in cultural heritage restoration.
In recent times, the use of natural and harmless products for the environment and restorer is taking place in the field of Cultural Heritage restoration. In this sense, wheat, rice and corn starches as adhesives, have suitable characteristics without toxicity risks. A new starch in this field, is the Kudzu, an almost pure compound (99.5% starch) that is processed by a natural way from a plant called Pueraria lobata. This is a preliminary study of the potential use of Kudzu starch for the restoration of Cultural Heritage, focusing, firstly, in its capacity as adhesive through a comparative evaluation with common starches. The accelerated aging process carried out proved that Kudzu ensures optimal chromatic behaviour. On the other hand, the main problem in starch paste is the biological colonization. The daidzein, a natural antimicrobial compound implicit in Kudzu starch, confirmed the resistance to microorganism in this preliminary approach. The evaluation of the adhesive capacity, and the reversibility of the starches, suggest that Kudzu starch is a valid adhesive in the field of paper restoration. Thus, the potential of this starch in the conservation of Cultural Heritage is evidenced and its use as cleaner, resistance to biological colonization and consolidant is promising.


Polymerization of Ethylene Oxide, Propylene Oxide, and Other Alkylene Oxides: Synthesis, Novel Polymer Architectures, and Bioconjugation.
The review summarizes current trends and developments in the polymerization of alkylene oxides in the last two decades since 1995, with a particular focus on the most important epoxide monomers ethylene oxide (EO), propylene oxide (PO), and butylene oxide (BO). Classical synthetic pathways, i.e., anionic polymerization, coordination polymerization, and cationic polymerization of epoxides (oxiranes), are briefly reviewed. The main focus of the review lies on more recent and in some cases metal-free methods for epoxide polymerization, i.e., the activated monomer strategy, the use of organocatalysts, such as N-heterocyclic carbenes (NHCs) and N-heterocyclic olefins (NHOs) as well as phosphazene bases. In addition, the commercially relevant double-metal cyanide (DMC) catalyst systems are discussed. Besides the synthetic progress, new types of multifunctional linear PEG (mf-PEG) and PPO structures accessible by copolymerization of EO or PO with functional epoxide comonomers are presented as well as complex branched, hyperbranched, and dendrimer like polyethers. Amphiphilic block copolymers based on PEO and PPO (Poloxamers and Pluronics) and advances in the area of PEGylation as the most important bioconjugation strategy are also summarized. With the ever growing toolbox for epoxide polymerization, a "polyether universe" may be envisaged that in its structural diversity parallels the immense variety of structural options available for polymers based on vinyl monomers with a purely carbon-based backbone.


The simultaneous effects of spatial and social networks on cholera transmission.
This study uses social network and spatial analytical methods simultaneously to understand cholera transmission in rural Bangladesh. Both have been used separately to incorporate context into health studies, but using them together is a new and recent approach. Data include a spatially referenced longitudinal demographic database consisting of approximately 200,000 people and a database of all laboratory-confirmed cholera cases from 1983 to 2003. A complete kinship-based network linking households is created, and distance matrices are also constructed to model spatial relationships. A spatial error-social effects model tested for cholera clustering in socially linked households while accounting for spatial factors. Results show that there was social clustering in five out of twenty-one years while accounting for both known and unknown environmental variables. This suggests that environmental cholera transmission is significant and social networks also influence transmission, but not as consistently. Simultaneous spatial and social network analysis may improve understanding of disease transmission.


Temperature Dependence of the Resonant Magnetoelectric Effect in Layered Heterostructures.
The dependence of the resonant direct magnetoelectric effect on temperature is studied experimentally in planar composite structures. Samples of rectangular shapes with dimensions of 5 mm × 20 mm employed ferromagnetic layers of either an amorphous (metallic glass) alloy or nickel with a thickness of 20-200 μm and piezoelectric layers of single crystalline langatate material or lead zirconate titanate piezoelectric ceramics with a thickness of 500 μm. The temperature of the samples was varied in a range between 120 and 390 K by blowing a gaseous nitrogen stream around them. It is shown that the effective characteristics of the magnetoelectric effect-such as the mechanical resonance frequency fr, the quality factor Q and the magnitude of the magnetoelectric coefficient αE at the resonance frequency-are contingent on temperature. The interrelations between the temperature changes of the characteristics of the magnetoelectric effect and the temperature variations of the following material parameters-Young's modulus Y, the acoustic quality factor of individual layers, the dielectric constant ε, the piezoelectric modulus d of the piezoelectric layer as well as the piezomagnetic coefficients λ(n) of the ferromagnetic layer-are established. The effect of temperature on the characteristics of the nonlinear magnetoelectric effect is observed for the first time. The results can be useful for designing magnetoelectric heterostructures with specified temperature characteristics, in particular, for the development of thermally stabilized magnetoelectric devices.


Acetabular retroversion is associated with osteoarthritis of the hip.
Primary osteoarthritis of the hip may have a structural basis. It was hypothesized that the radiographic appearance of acetabular retroversion could be created by altering the morphologic features of the acetabular walls, and that acetabular retroversion, as defined on an anteroposterior radiograph of the pelvis, is associated with osteoarthritis of the hip. A model pelvis was used to simulate normal, augmented, deficient, and rotated walls of the acetabulum, and radiographs were taken to compare the projections of the modified acetabular walls with the known plain radiographic appearance of a retroverted acetabulum. One hundred thirty-one good quality anteroposterior radiographs of the pelvis taken before total hip arthroplasty for idiopathic hip osteoarthritis were compared with 99 good quality radiographs taken for nonorthopaedic reasons. The prevalence of radiographic acetabular retroversion is 20% among patients with idiopathic hip osteoarthritis and 5% among the general population. The appearance of acetabular retroversion on an anteoroposterior radiograph of the pelvis is created by deficiency of the posterior wall of the acetabulum. There is a statistically significant association between radiographic acetabular retroversion and hip osteoarthritis. These findings have applicability to understanding the mechanical etiology of hip osteoarthritis, and to surgical technique during periacetabular osteotomy and total hip arthroplasty.


Function and dynamics of PKD2 in Chlamydomonas reinhardtii flagella.
To analyze the function of ciliary polycystic kidney disease 2 (PKD2) and its relationship to intraflagellar transport (IFT), we cloned the gene encoding Chlamydomonas reinhardtii PKD2 (CrPKD2), a protein with the characteristics of PKD2 family members. Three forms of this protein (210, 120, and 90 kD) were detected in whole cells; the two smaller forms are cleavage products of the 210-kD protein and were the predominant forms in flagella. In cells expressing CrPKD2-GFP, about 10% of flagellar CrPKD2-GFP was observed moving in the flagellar membrane. When IFT was blocked, fluorescence recovery after photobleaching of flagellar CrPKD2-GFP was attenuated and CrPKD2 accumulated in the flagella. Flagellar CrPKD2 increased fourfold during gametogenesis, and several CrPKD2 RNA interference strains showed defects in flagella-dependent mating. These results suggest that the CrPKD2 cation channel is involved in coupling flagellar adhesion at the beginning of mating to the increase in flagellar calcium required for subsequent steps in mating.


Pulmonary chemoreflex sensitivity is enhanced by prostaglandin E2 in anesthetized rats.
Stimulation of vagal pulmonary C-fiber afferents by chemical irritants is believed to be responsible for eliciting the pulmonary chemoreflex (apnea, bradycardia, and hypotension). This study was carried out in anesthetized Sprague-Dawley rats to determine whether the pulmonary chemoreflex was altered by prostaglandin E2 (PGE2), which is one of the major inflammatory mediators in the lungs and is known to enhance the sensitivity of C-fiber afferents in several other organ systems. Capsaicin injected at a dose just above the stimulation threshold (0.25 or 0.5 microgram/kg i.v.) elicited a very mild respiratory and cardiovascular depression. In sharp contrast, during a constant infusion of PGE2 (1.5 micrograms.kg-1.min-1 i.v.), the same dose of capsaicin triggered a long apnea, with the expiratory duration reaching 843% of the baseline expiratory duration, accompanied by intense bradycardia and hypotension. Similarly, the pulmonary chemoreflex response elicited by a bolus injection of phenyl biguanide (1 or 2 micrograms/kg i.v.) was also greatly augmented by PGE2. These enhanced responses were completely abolished, by a perineural capsaicin treatment of both cervical vagi to selectively block the conduction of C fibers, suggesting the involvement of these afferents. Electrophysiological recording of pulmonary C-fiber afferent activity further supported our conclusion that the sensitivity of these sensory endings to capsaicin challenge was potentiated by PGE2.


Desulfocarbo indianensis gen. nov., sp. nov., a benzoate-oxidizing, sulfate-reducing bacterium isolated from water extracted from a coal bed.
A novel, strictly anaerobic, sulfate-reducing bacterium, designated strain SCBM(T), was isolated from water extracted from a coal bed in Indiana, USA. The isolate was characterized by a polyphasic taxonomic approach that included phenotypic and genotypic characterizations. Cells of strain SCBM(T) were vibrio-shaped, polarly flagellated, Gram-negative, motile, oxidase-negative and weakly catalase-positive. Growth of strain SCBM(T) was observed at NaCl concentrations ranging from 0 to 300 mM. However, no growth was observed when 1 M or more NaCl was present. Growth was observed at 16-37 °C, with optimal growth at 30 °C. The optimum pH for growth was 7, although growth was observed from pH 6.5 to 8. The doubling time under optimal growth conditions (30 °C, pH 7, 2.5 mM benzoate, 14 mM sulfate) was 2.7 days. Bicarbonate, HEPES, PIPES and MES were effective buffers for growth of strain SCBM(T), but citrate inhibited growth. When sulfate was provided as the electron acceptor, strain SCBM(T) grew autotrophically with hydrogen as the electron donor and heterotrophically on benzoate, formate, acetate, pyruvate, butyrate, fumarate, succinate and palmitate. None of the substrates tested supported fermentative growth. Thiosulfate and sulfate were used as electron acceptors coupled to benzoate oxidation, but sulfite, elemental sulfur, DMSO, anthraquinone 2,6-disulfonate, nitrate, nitrite, ferric citrate, hydrous iron oxide and oxygen were not. The G+C content of genomic DNA was 62.5 mol%. The major cellular fatty acids were anteiso-C(15 : 0) and C(18 : 1)ω7c. Phylogenetic analysis based on 16S rRNA gene sequencing placed strain SCBM(T) into a distinct lineage within the class Deltaproteobacteria. The closest, cultivated phylogenetic relative of strain SCBM(T) was Desulfarculus baarsii DSM 2075(T), with only 91.7% 16S rRNA gene sequence identity. On the basis of phenotypic and genotypic analyses, strain SCBM(T) represents a novel genus and species of sulfate-reducing bacteria, for which the name Desulfocarbo indianensis gen. nov., sp. nov. is proposed. The type strain of Desulfocarbo indianensis is SCBM(T) ( = DSM 28127(T) = JCM 19826(T)). Desulfocarbo is the second genus of the order Desulfarculales.


Expedited Safety Reporting to Sponsors Through the Implementation of an Alert System for Clinical Trial Management at an Academic Medical Center: Retrospective Design Study.
Early detection or notification of adverse event (AE) occurrences during clinical trials is essential to ensure patient safety. Clinical trials take advantage of innovative strategies, clinical designs, and state-of-the-art technologies to evaluate efficacy and safety, however, early awareness of AE occurrences by investigators still needs to be systematically improved. This study aimed to build a system to promptly inform investigators when clinical trial participants make unscheduled visits to the emergency room or other departments within the hospital. We developed the Adverse Event Awareness System (AEAS), which promptly informs investigators and study coordinators of AE occurrences by automatically sending text messages when study participants make unscheduled visits to the emergency department or other clinics at our center. We established the AEAS in July 2015 in the clinical trial management system. We compared the AE reporting timeline data of 305 AE occurrences from 74 clinical trials between the preinitiative period (December 2014-June 2015) and the postinitiative period (July 2015-June 2016) in terms of three AE awareness performance indicators: onset to awareness, awareness to reporting, and onset to reporting. A total of 305 initial AE reports from 74 clinical trials were included. All three AE awareness performance indicators were significantly lower in the postinitiative period. Specifically, the onset-to-reporting times were significantly shorter in the postinitiative period (median 1 day [IQR 0-1], mean rank 140.04 [SD 75.35]) than in the preinitiative period (median 1 day [IQR 0-4], mean rank 173.82 [SD 91.07], P≤.001). In the phase subgroup analysis, the awareness-to-reporting and onset-to-reporting indicators of phase 1 studies were significantly lower in the postinitiative than in the preinitiative period (preinitiative: median 1 day, mean rank of awareness to reporting 47.94, vs postinitiative: median 0 days, mean rank of awareness to reporting 35.75, P=.01; and preinitiative: median 1 day, mean rank of onset to reporting 47.4, vs postinitiative: median 1 day, mean rank of onset to reporting 35.99, P=.03). The risk-level subgroup analysis found that the onset-to-reporting time for low- and high-risk studies significantly decreased postinitiative (preinitiative: median 4 days, mean rank of low-risk studies 18.73, vs postinitiative: median 1 day, mean rank of low-risk studies 11.76, P=.02; and preinitiative: median 1 day, mean rank of high-risk studies 117.36, vs postinitiative: median 1 day, mean rank of high-risk studies 97.27, P=.01). In particular, onset to reporting was reduced more in the low-risk trial than in the high-risk trial (low-risk: median 4-0 days, vs high-risk: median 1-1 day). We demonstrated that a real-time automatic alert system can effectively improve safety reporting timelines. The improvements were prominent in phase 1 and in low- and high-risk clinical trials. These findings suggest that an information technology-driven automatic alert system effectively improves safety reporting timelines, which may enhance patient safety.


Robust video object cosegmentation.
With ever-increasing volumes of video data, automatic extraction of salient object regions became even more significant for visual analytic solutions. This surge has also opened up opportunities for taking advantage of collective cues encapsulated in multiple videos in a cooperative manner. However, it also brings up major challenges, such as handling of drastic appearance, motion pattern, and pose variations, of foreground objects as well as indiscriminate backgrounds. Here, we present a cosegmentation framework to discover and segment out common object regions across multiple frames and multiple videos in a joint fashion. We incorporate three types of cues, i.e., intraframe saliency, interframe consistency, and across-video similarity into an energy optimization framework that does not make restrictive assumptions on foreground appearance and motion model, and does not require objects to be visible in all frames. We also introduce a spatio-temporal scale-invariant feature transform (SIFT) flow descriptor to integrate across-video correspondence from the conventional SIFT-flow into interframe motion flow from optical flow. This novel spatio-temporal SIFT flow generates reliable estimations of common foregrounds over the entire video data set. Experimental results show that our method outperforms the state-of-the-art on a new extensive data set (ViCoSeg).


Transcriptome profiling of bovine milk oligosaccharide metabolism genes using RNA-sequencing.
This study examines the genes coding for enzymes involved in bovine milk oligosaccharide metabolism by comparing the oligosaccharide profiles with the expressions of glycosylation-related genes. Fresh milk samples (n = 32) were collected from four Holstein and Jersey cows at days 1, 15, 90 and 250 of lactation and free milk oligosaccharide profiles were analyzed. RNA was extracted from milk somatic cells at days 15 and 250 of lactation (n = 12) and gene expression analysis was conducted by RNA-Sequencing. A list was created of 121 glycosylation-related genes involved in oligosaccharide metabolism pathways in bovine by analyzing the oligosaccharide profiles and performing an extensive literature search. No significant differences were observed in either oligosaccharide profiles or expressions of glycosylation-related genes between Holstein and Jersey cows. The highest concentrations of free oligosaccharides were observed in the colostrum samples and a sharp decrease was observed in the concentration of free oligosaccharides on day 15, followed by progressive decrease on days 90 and 250. Ninety-two glycosylation-related genes were expressed in milk somatic cells. Most of these genes exhibited higher expression in day 250 samples indicating increases in net glycosylation-related metabolism in spite of decreases in free milk oligosaccharides in late lactation milk. Even though fucosylated free oligosaccharides were not identified, gene expression indicated the likely presence of fucosylated oligosaccharides in bovine milk. Fucosidase genes were expressed in milk and a possible explanation for not detecting fucosylated free oligosaccharides is the degradation of large fucosylated free oligosaccharides by the fucosidases. Detailed characterization of enzymes encoded by the 92 glycosylation-related genes identified in this study will provide the basic knowledge for metabolic network analysis of oligosaccharides in mammalian milk. These candidate genes will guide the design of a targeted breeding strategy to optimize the content of beneficial oligosaccharides in bovine milk.