Lytic versus stimulatory synapse in cytotoxic T lymphocyte/target cell interaction: manifestation of a dual activation threshold.
Activation of biological functions in T lymphocytes is determined by the molecular dynamics occurring at the T cell/opposing cell interface. In the present study, a central question of cytotoxic T lymphocyte (CTL) biology was studied at the single-cell level: can two distinct activation thresholds for cytotoxicity and cytokine production be explained by intercellular molecular dynamics between CTLs and targets? In this study, we combine morphological approaches with numerical analysis, which allows us to associate specific patterns of calcium mobilization with different biological responses. We show that CTLs selectively activated to cytotoxicity lack a mature immunological synapse while exhibiting a low threshold polarized secretion of lytic granules and spike-like patterns of calcium mobilization. This finding is contrasted by fully activated CTLs, which exhibit a mature immunological synapse and smooth and sustained calcium mobilization. Our results indicate that intercellular molecular dynamics and signaling characteristics allow the definition of two activation thresholds in individual CTLs: one for polarized granule secretion (lytic synapse formation) and the other for cytokine production (stimulatory synapse formation).


Aortorenal arterial autografts: long-term assessment.
Ninety-four iliac arterial autografts were placed in 86 hypertensive patients (age range, 7 to 57 years) during a 16-year period for renal artery reconstruction of nonatherosclerotic lesions. Ten patients were children, and nine patients had a solitary kidney. Ex vivo repair was used in 21 patients. There were no early postoperative deaths; one patient died three months postoperatively of unrelated disease. Two grafts occluded in the early postoperative period. Follow-up of one to 16 years (mean, 5.6 years) was available in 77 patients (93%). Forty-one patients underwent late arteriography at an average of 5.7 years postoperatively. Fifty-one patients (66%) were regarded as cured of their hypertension, the conditions of 25 patients (32%) improved, and the condition of one patient (1.2%) was unchanged. There were no late autograft occlusions. Atherosclerotic anastomotic stenosis developed in one autograft 13 years postoperatively and was relieved by reoperation. Dilation of one autograft was observed seven years postoperatively but did not increase on subsequent arteriograms. In five children, follow-up angiograms showed that the size of the autograft paralleled that of the contralateral renal artery. These results support the preferential use of an autograft during reconstruction of nonatherosclerotic lesions that cause renovascular hypertension.


Immunogenicity of panitumumab in combination chemotherapy clinical trials.
Panitumumab is a fully human antibody against the epidermal growth factor receptor that is indicated for the treatment of metastatic colorectal cancer (mCRC) after disease progression on standard chemotherapy. The purpose of this analysis was to examine the immunogenicity of panitumumab and to evaluate the effect of anti-panitumumab antibodies on pharmacokinetic and safety profiles in patients with mCRC receiving panitumumab in combination with oxaliplatin- or irinotecan-based chemotherapies. Three validated assays (two screening immunoassays and a neutralizing antibody bioassay) were used to detect the presence of anti-panitumumab antibodies in serum samples collected from patients enrolled in four panitumumab combination chemotherapy clinical trials. The impact of anti-panitumumab antibodies on pharmacokinetic and safety profiles was analyzed using population pharmacokinetic analysis and descriptive statistics, respectively. Of 1124 patients treated with panitumumab in combination with oxaliplatin- or irinotecan-based chemotherapy with postbaseline samples available for testing, 20 (1.8%) patients developed binding antibodies and 2 (0.2%) developed neutralizing antibodies. The incidence of anti-panitumumab antibodies was similar in patients with tumors expressing wild-type or mutant KRAS and in patients receiving oxaliplatin- or irinotecan-based chemotherapies. No evidence of an altered pharmacokinetic or safety profile was found in patients who tested positive for anti-panitumumab antibodies. The immunogenicity of panitumumab in the combination chemotherapy setting was infrequent and similar to the immunogenicity observed in the monotherapy setting. Panitumumab immunogenicity did not appear to alter pharmacokinetic or safety profiles. This low rate of immunogenicity may be attributed to the fully human nature of panitumumab.


Did tool-use evolve with enhanced physical cognitive abilities?
The use and manufacture of tools have been considered to be cognitively demanding and thus a possible driving factor in the evolution of intelligence. In this study, we tested the hypothesis that enhanced physical cognitive abilities evolved in conjunction with the use of tools, by comparing the performance of naturally tool-using and non-tool-using species in a suite of physical and general learning tasks. We predicted that the habitually tool-using species, New Caledonian crows and Galápagos woodpecker finches, should outperform their non-tool-using relatives, the small tree finches and the carrion crows in a physical problem but not in general learning tasks. We only found a divergence in the predicted direction for corvids. That only one of our comparisons supports the predictions under this hypothesis might be attributable to different complexities of tool-use in the two tool-using species. A critical evaluation is offered of the conceptual and methodological problems inherent in comparative studies on tool-related cognitive abilities.


Apolipoprotein E epsilon 4 is a determinant for Alzheimer-type pathologic features in tauopathies, synucleinopathies, and frontotemporal degeneration.
To determine if apolipoprotein E epsilon 4 influences the frequency of Alzheimer-type pathologic features in tauopathies, synucleinopathies, and frontotemporal degeneration and to determine if the frequency of Alzheimer-type pathologic features in synucleinopathies is similar to the frequency of such features in tauopathies and frontotemporal degeneration. A total of 285 patients with pathologically proven neurodegenerative disorders, including diffuse and transitional Lewy body disease, frontotemporal degeneration, progressive supranuclear palsy, corticobasal degeneration, and multiple system atrophy, with a mean age of 75.1 +/- 9.3 years, were suitable for genetic and pathological analysis. Disorders were grouped as tauopathies (progressive supranuclear palsy and corticobasal degeneration), synucleinopathies (Lewy body disease and multiple system atrophy), and frontotemporal degeneration. Braak neurofibrillary tangle staging and quantitative scores of senile plaques were used to determine the degree of concomitant Alzheimer-type pathologic features in each case, and apolipoprotein E genotype was determined from DNA isolated from frozen brain tissue. The relationship of apolipoprotein E epsilon 4 to Alzheimer-type pathologic features was determined. Across all neurodegenerative disorders, apolipoprotein E epsilon 4 and older age independently predicted the co-occurrence of Alzheimer-type pathologic features (P<.001), whereas female sex had a lesser effect (P = .03). When divided into the 3 subgroups (tauopathies, synucleinopathies, and frontotemporal degeneration), apolipoprotein E epsilon 4 had a similar effect, whereas older age and female sex were less predictive. There was a significant difference between the frequency of Alzheimer-type pathologic features in synucleinopathies and the frequency of such features in tauopathies and frontotemporal degeneration (P<.001 for both). The frequency of apolipoprotein E epsilon 4 allele was not significantly different among the 3 groups. Apolipoprotein E epsilon 4, independent of older age and sex, contributes to the co-occurrence of Alzheimer-type pathologic features in tauopathies, synucleinopathies, and frontotemporal degeneration, but this does not explain why Alzheimer-type pathologic features are significantly more likely to coexist with synucleinopathies than with either tauopathies or frontotemporal degeneration.


Effects of high-intensity intermittent running exercise in overweight children.
This study examined the effects of a 6-week intermittent exercise training, at different intensities, on body composition, functional walking and aerobic endurance in overweight children. Forty-eight overweight children (age: 10.4 ± 0.9 years) were randomly assigned to either intervention or control group. Lower and higher intensity intermittent exercise groups (LIIE and HIIE) performed intermittent running three times a week. LIIE performed more intervals at a lower intensity [16 intervals at 100% of individual maximal aerobic speed (MAS), 8 minutes in total], and HIIE performed fewer intervals at a higher intensity (12 intervals at 120% of MAS, 6 minutes in total). Each interval consisted of a 15-second run at the required speed, followed by a 15-second passive recovery. After 6 weeks, HIIE had a significantly (p < 0.05) higher percentage reduction in sum of skinfolds (i.e. calf and triceps), and significantly (p < 0.05) fewer steps during the functional obstacle performance, as compared with LIIE and control group. Significant improvement (p < 0.05) was found in intermittent aerobic endurance for HIIE as compared to the control group. Higher intensity intermittent training is an effective and time-efficient intervention for improving body composition, functional walking and aerobic endurance in overweight children.


An unusual cause of peritonitis in peritoneal dialysis patients: Pantoea agglomerans.
Peritonitis is a serious infection and early diagnosis and treatment is mandatory. A variety of microorganisms are identified in these cases and during recent years a new one was included, Pantoea agglomerans. In this case report, a female patient on continuous ambulatory peritoneal dialysis therapy with a peritonitis episode caused by this organism is described. The source of infection was thought to be due to contact of catheter with non-sterile surfaces. In microbiologic culture, this organism was identified and the patient successfully treated with a three week course of gentamicin therapy. The number of reported cases with this organism has increased in last years and various infection localizations and clinical progress patterns have been identified. In peritoneal dialysis patients presenting with peritonitis, this organism must be kept in mind.


Pseudorabies virus gene encoding glycoprotein gIII is not essential for growth in tissue culture.
We have established that in the Becker strain of pseudorabies virus (PRV), the glycoprotein gIII gene is not essential for growth in cell culture. This was accomplished by construction and analysis of viral mutants containing two defined deletion mutations affecting the gIII gene. These mutations were first constructed in vitro and introduced into Escherichia coli expression plasmids to verify structure and protein production. Each mutation was then crossed onto PRV by cotransfection of plasmid DNA and parental viral DNA by using gIII-specific monoclonal antibodies as selective and screening reagents. One resultant virus strain, PRV-2, contained an in-frame deletion of a 402-base-pair (bp) SacI fragment contained within the gIII gene. Another virus strain, PRV-10, contained a deletion of a 1,480-bp XhoI fragment removing 230 bp of the upstream, putative transcriptional control sequences and 87% of the gIII coding sequence. The deletion mutants were compared with parental virus by analysis of virion DNA, gIII specific RNA, and proteins reacting with gIII specific antibodies. Upon infection of PK15 cells, the deletion mutants did not produce any proteins that reacted with two gIII specific monoclonal antibodies. However, two species of truncated glycosylated proteins were observed in PRV-2 infected cells that reacted with antiserum raised against bacterially produced gIII protein. PRV-10 produced no detectable gIII-specific RNA or protein. PRV-10 could be propagated without difficulty in tissue culture. Virus particles lacking gIII were indistinguishable from parental PRV virus particles by analysis of infected-cell thin sections in the electron microscope. We therefore conclude that expression of the gIII gene was not absolutely essential for PRV growth in tissue culture.


Expression of alternative oxidase in Drosophila ameliorates diverse phenotypes due to cytochrome oxidase deficiency.
Mitochondrial dysfunction is a significant factor in human disease, ranging from systemic disorders of childhood to cardiomyopathy, ischaemia and neurodegeneration. Cytochrome oxidase, the terminal enzyme of the mitochondrial respiratory chain, is a frequent target. Lower eukaryotes possess alternative respiratory-chain enzymes that provide non-proton-translocating bypasses for respiratory complexes I (single-subunit reduced nicotinamide adenine dinucleotide dehydrogenases, e.g. Ndi1 from yeast) or III + IV [alternative oxidase (AOX)], under conditions of respiratory stress or overload. In previous studies, it was shown that transfer of yeast Ndi1 or Ciona intestinalis AOX to Drosophila was able to overcome the lethality produced by toxins or partial knockdown of complex I or IV. Here, we show that AOX can provide a complete or substantial rescue of a range of phenotypes induced by global or tissue-specific knockdown of different cIV subunits, including integral subunits required for catalysis, as well as peripheral subunits required for multimerization and assembly. AOX was also able to overcome the pupal lethality produced by muscle-specific knockdown of subunit CoVb, although the rescued flies were short lived and had a motility defect. cIV knockdown in neurons was not lethal during development but produced a rapidly progressing locomotor and seizure-sensitivity phenotype, which was substantially alleviated by AOX. Expression of Ndi1 exacerbated the neuronal phenotype produced by cIV knockdown. Ndi1 expressed in place of essential cI subunits produced a distinct residual phenotype of delayed development, bang sensitivity and male sterility. These findings confirm the potential utility of alternative respiratory chain enzymes as tools to combat mitochondrial disease, while indicating important limitations thereof.


Perinatal characteristics and obstetric complications as risk factors for asthma, allergy and eczema at the age of 6 years.
Considerable effort has been put into identifying early determinants for atopic disorders. Many studies have evaluated the role of fetal development and obstetric complications. However, the results are not unequivocal. To assess the relationship between perinatal characteristics and obstetric complications, and the presence of reported current asthma, allergy and eczema at the age of 6 years in the framework of a previously conducted study. Seven hundred families in the Netherlands with index children born in 1988-1990 were retrospectively selected. Data were extracted from the Municipal Health Service's records of health examinations of these children and their siblings. These examinations were carried out at the age of 6 years. The records contained data on reported atopic disorders and perinatal characteristics. Gestational age was inversely related to the risk of asthma (P for trend: 0.03). Children with low birth weight tended to have a lower risk of any allergy, albeit not significant (P=0.07). However, no link was found between neonatal head circumference and atopic disorders. The ratio of neonatal head circumference to birth weight was positively associated with the risk of atopic disorders, especially with the risk of asthma (odds ratio (OR)=1.87; 95% confidence interval (CI(95%))=[1.11, 3.15]). Vacuum extraction was a risk factor for allergy (OR=1.84, CI(95%)=[1.03, 3.28]), but not for asthma. Induced labour was positively associated with the risk of inhalant allergy (OR=2.22, CI(95%)=[1.09, 4.51]) and, to a lesser extent, asthma (OR=1.72, CI(95%)=[0.95, 3.10]). For caesarean section and forcipal extraction there were no such relationships. Prematurity is a risk factor for asthma reported at 6 years. A high ratio of head circumference to birth weight is a risk factor for any atopic disorder. Vacuum extraction was associated with a higher risk of allergy, and induced labour is a risk factor for inhalant allergy. All results should be viewed with the possibility of residual confounding.


Focused metabolomic profiling in the drug development process: advances from lipid profiling.
The highly parallel analytical technologies comprising 'omics promised to dramatically improve drug development efficiency by increasing knowledge and improving decision-making capabilities. On this point, the 'omics have largely been a disappointment. The major reason genomics, transcriptomics and proteomics fail to improve decision making capabilities is that they produce so many false positive results that it is difficult to be sure that findings are valid. Metabolomics is not immune to this problem but, when practiced effectively, the technology can reliably produce knowledge to aid in decision making. In particular, focused metabolomics platforms - those that restrict their target analytes to those measured well by the technology - can produce data with properties that maximize sensitivity and minimize the false discovery problem. The most developed focused metabolomics area is lipid profiling.


Sublingual nitroglycerin: improving patient compliance with a demonstration dose.
Forty-four patients were studied for compliance in the use of their first prescription for sublingual nitroglycerin. Fourteen HMO physicians participated in this randomized, prospective study by administering the first dose to half the patients during the office visit in which the diagnosis of angina pectoris was made. No other changes were made in the physicians' customary methods of diagnosing and treating angina pectoris. Patients who received the demonstration dose were significantly more likely to have used the sublingual nitroglycerin at least once before their second visit (75 percent of the study group compared with 44 percent of the control group). There was no difference between the groups in having the drug with them at the second visit. While serious reactions to sublingual nitroglycerin are rare, 6 patients experienced symptoms from the first dose, including 1 patient who fainted. The results suggest that compliance in using the initial prescription for sublingual nitroglycerin can be improved when the physician supervises the first dose.


Angiopoietin-like 4 (Angptl4): A glucocorticoid-dependent gatekeeper of fatty acid flux during fasting.
Angiopoietin-like 4 (Angptl4) is a secreted protein modulating triacylglycerol homeostasis. Its transcription is induced by glucocorticoids, which act to elevate circulating Angptl4 levels during fasting. In investigating the role of Angptl4 in glucocorticoid action, we identified that in addition to its known ability to inhibit lipoprotein lipase, Angptl4 stimulates intracellular adipocyte lipolysis. Fatty acid release by murine adipocytes following fasting or treatment with glucocorticoids or catecholamines is highly Angptl4-dependent. In fact, Angptl4 can directly stimulate cAMP-dependent PKA signaling and lipolysis when added to adipocytes. Here, we detail this novel Angptl4-dependent lipolytic regulatory mechanism and discuss its physiological and therapeutic implications.


Intermittent administration of inhaled tobramycin in patients with cystic fibrosis. Cystic Fibrosis Inhaled Tobramycin Study Group.
We conducted two multicenter, double-blind, placebo-controlled trials of intermittent administration of inhaled tobramycin in patients with cystic fibrosis and Pseudomonas aeruginosa infection. A total of 520 patients (mean age, 21 years) were randomly assigned to receive either 300 mg of inhaled tobramycin or placebo twice daily for four weeks, followed by four weeks with no study drug. Patients received treatment or placebo in three on-off cycles for a total of 24 weeks. The end points included pulmonary function, the density of P. aeruginosa in sputum, and hospitalization. The patients treated with inhaled tobramycin had an average increase in forced expiratory volume in one second (FEV1) of 10 percent at week 20 as compared with week 0, whereas the patients receiving placebo had a 2 percent decline in FEV1 (P<0.001). In the tobramycin group, the density of P. aeruginosa decreased by an average of 0.8 log10 colony-forming units (CFU) per gram of expectorated sputum from week 0 to week 20, as compared with an increase of 0.3 log10 CFU per gram in the placebo group (P<0.001). The patients in the tobramycin group were 26 percent (95 percent confidence interval, 2 to 43 percent) less likely to be hospitalized than those in the placebo group. Inhaled tobramycin was not associated with detectable ototoxic or nephrotoxic effects or with accumulation of the drug in serum. The proportion of patients with P. aeruginosa isolates for which the minimal inhibitory concentration of tobramycin was 8 microg per milliliter or higher increased from 25 percent at week 0 to 32 percent at week 24 in the tobramycin group, as compared with a decrease from 20 percent at week 0 to 17 percent at week 24 in the placebo group. In a 24-week study of patients with cystic fibrosis, intermittent administration of inhaled tobramycin was well tolerated and improved pulmonary function, decreased the density of P. aeruginosa in sputum, and decreased the risk of hospitalization.


Inhibition of NADPH cytochrome P450 reductase by the model sulfur mustard vesicant 2-chloroethyl ethyl sulfide is associated with increased production of reactive oxygen species.
Inhalation of vesicants including sulfur mustard can cause significant damage to the upper airways. This is the result of vesicant-induced modifications of proteins important in maintaining the integrity of the lung. Cytochrome P450s are the major enzymes in the lung mediating detoxification of sulfur mustard and its metabolites. NADPH cytochrome P450 reductase is a flavin-containing electron donor for cytochrome P450. The present studies demonstrate that the sulfur mustard analog, 2-chloroethyl ethyl sulfide (CEES), is a potent inhibitor of human recombinant cytochrome P450 reductase, as well as native cytochrome P450 reductase from liver microsomes of saline and beta-naphthoflavone-treated rats, and cytochrome P450 reductase from type II lung epithelial cells. Using rat liver microsomes from beta-naphthoflavone-treated rats, CEES was found to inhibit CYP 1A1 activity. This inhibition was overcome by microsomal cytochrome P450 reductase from saline-treated rats, which lack CYP 1A1 activity, demonstrating that the CEES inhibitory activity was selective for cytochrome P450 reductase. Cytochrome P450 reductase also generates reactive oxygen species (ROS) via oxidation of NADPH. In contrast to its inhibitory effects on the reduction of cytochrome c and CYP1A1 activity, CEES was found to stimulate ROS formation. Taken together, these data demonstrate that sulfur mustard vesicants target cytochrome P450 reductase and that this effect may be an important mechanism mediating oxidative stress and lung injury.


Effects of oxygen and glucose deprivation on vasoactivity in isolated bovine middle cerebral arteries.
The effects of oxygen and glucose deprivation on vasoactivity were investigated using helical strips of bovine middle cerebral artery. Hypoxia, created by reducing the PO2 of the bath, or oxidative inhibition with 2,4 dinitrophenol (DNP) or sodium azide, significantly reduced contractions induced by serotonin. Normal tonic contractions induced with fresh and aged whole blood, or 5-HT became phasic and quickly relaxed to baseline in a hypoxic environment. Glucose elimination from the Krebs medium, or the inhibition of the glycolytic pathway with iodoacetic acid (IAA), did not significantly reduce serotonin-induced contractions. However, contractions were inhibited more with the combination of oxygen and glucose deprivation, or DNP + IAA, than with oxygen deprivation alone. Efforts to produce rigor in this preparation by oxygen/substrate reduction or metabolic inhibition were unsuccessful. Tonic contractions induced by 70 mM potassium became phasic as the Ca++ concentration was reduced. Contractions resulting from the readdition of Ca++ to arteries exposed to calcium-free high potassium solution were significantly reduced in the presence of oxidative and/or glycolytic inhibitors. The uptake of 45Ca++, as measured by the lanthanum technique, decreased as the bath PO2 was reduced in both serotonin stimulated and unstimulated arteries. Glucose deprivation alone did not affect 45Ca++ uptake. This study suggests that hypoxia has a direct inhibitory affect on cerebral vasoactivity mediated by reductions in sarcoplasmic Ca++ uptake.


A novel cyclic peptide (Naturido) modulates glia-neuron interactions in vitro and reverses ageing-related deficits in senescence-accelerated mice.
The use of agents that target both glia and neurons may represent a new strategy for the treatment of ageing disorders. Here, we confirmed the presence of the novel cyclic peptide Naturido that originates from a medicinal fungus (Isaria japonica) grown on domestic silkworm (Bombyx mori). We found that Naturido significantly enhanced astrocyte proliferation and activated the single copy gene encoding the neuropeptide VGF and the neuron-derived NGF gene. The addition of the peptide to the culture medium of primary hippocampal neurons increased dendrite length, dendrite number and axon length. Furthermore, the addition of the peptide to primary microglial cultures shifted CGA-activated microglia towards anti-inflammatory and neuroprotective phenotypes. These findings of in vitro glia-neuron interactions led us to evaluate the effects of oral administration of the peptide on brain function and hair ageing in senescence-accelerated mice (SAMP8). In vivo analyses revealed that spatial learning ability and hair quality were improved in Naturido-treated mice compared with untreated mice, to the same level observed in the normal ageing control (SAMR1). These data suggest that Naturido may be a promising glia-neuron modulator for the treatment of not only senescence, but also Alzheimer's disease and other neurodegenerative diseases.


Allogeneic tooth transplantation in the dog.
Allogeneic tooth transplantation was evaluated as a functional and aesthetic treatment for dental fracture in the dog. Of 7 dogs that received tooth transplants, 5 were research animals and 2 were clinical patients. Canine teeth were transplanted immediately after extraction in the research dogs. Endodontic therapy had been performed on the affected canine tooth of one clinical patient. The other clinical patient had bilateral maxillary canine fractures 2 months earlier. One of the research dog transplants failed at 3 weeks as a result of improper surgical technique. Four of the research dogs had a solid implant for 18 months, after which time the dogs were euthanatized serially. All transplanted teeth were anchored firmly into the alveoli, but were nonviable. Root resorption, with bone replacement, was first noticed at 24 months. The transplanted tooth in the first clinical patient remained functional for 3 months, after which time the tooth was fractured. The right canine transplant in the second clinical patient failed by 3 months, probably because of preexisting periapical inflammation. The left transplanted tooth remains stable at 38 months. It was concluded that allogeneic tooth transplantation may have merit as a rapid and inexpensive method for replacement of fractured teeth in the dog. Function is compromised gradually as a result of root resorption and ankylosis, with tooth fracture likely to occur after 2 years.


Tracing Origins of the Salmonella Bareilly Strain Causing a Food-borne Outbreak in the United States.
Using a novel combination of whole-genome sequencing (WGS) analysis and geographic metadata, we traced the origins of Salmonella Bareilly isolates collected in 2012 during a widespread food-borne outbreak in the United States associated with scraped tuna imported from India. Using next-generation sequencing, we sequenced the complete genome of 100 Salmonella Bareilly isolates obtained from patients who consumed contaminated product, from natural sources, and from unrelated historically and geographically disparate foods. Pathogen genomes were linked to geography by projecting the phylogeny on a virtual globe and produced a transmission network. Phylogenetic analysis of WGS data revealed a common origin for outbreak strains, indicating that patients in Maryland and New York were infected from sources originating at a facility in India. These data represent the first report fully integrating WGS analysis with geographic mapping and a novel use of transmission networks. Results showed that WGS vastly improves our ability to delimit the scope and source of bacterial food-borne contamination events. Furthermore, these findings reinforce the extraordinary utility that WGS brings to global outbreak investigation as a greatly enhanced approach to protecting the human food supply chain as well as public health in general.


Pheochromocytoma after Cesarean Section.
Pheochromocytoma is a catecholamine-producing tumor. There are a very few reported cases of clinical pheochromocytoma. Here, we report a 27-year-old woman para 1 live 1 with chief complaint of headache, confusion, nausea, and vomiting 2 days after cesarean section. She was anxious and had palpitation. On physical examination, fever, tachycardia, tachypnea, high blood pressure, and right thyroid nodule were found. She was managed as pregnancy-induced hypertension at first. In laboratory data, epinephrine, norepinephrine, metanephrine, normetanephrine, and vanillylmandelic acid were increased in 24 h urine collection. An adrenal mass was detected in abdominal computed tomography. Regarding clinical and paraclinical findings, pheochromocytoma was diagnosed. The patient received medical treatment, but it was not effective; hence, she underwent adrenalectomy.


Comparison of hemodynamic measurements from invasive and noninvasive monitoring during early resuscitation.
Measurements obtained from the insertion of a pulmonary artery catheter (PAC) in critically ill and/or injured patients have traditionally assisted with resuscitation efforts. However, with the recent utilization of ultrasound in the intensive care unit setting, transthoracic echocardiography (TTE) has gained popularity. The purpose of this study is to compare serial PAC and TTE measurements and document levels of serum biomarkers during resuscitation. Over a 25-month period, critically ill and/or injured patients admitted to a Level I adult trauma center were enrolled in this 48-hour intensive care unit study. Serial PAC and TTE measurements were obtained every 12 hours (total = 5 points/patient). Serial levels of lactate, Δ base, troponin-1, and B-type natriuretic peptide were obtained. Pearson correlation coefficient and intraclass correlation (ICC) assessed relationship and agreement, respectively, between PAC and TTE measures of cardiac output (CO) and stroke volume (SV). Analysis of variance with post hoc pairwise determined differences over time. Of the 29 patients, 69% were male, with a mean age of 47.4 years ± 19.5 years and 79.3% survival. Of these, 25 of 29 were trauma with a mean Injury Severity Score of 23.5 ± 10.7. CO from PAC and TTE was significantly related (Pearson correlations, 0.57-0.64) and agreed with moderate strength (ICC, 0.66-0.70). SV from PAC and TTE was significantly related (Pearson correlations, 0.40-0.58) and agreed at a weaker level (ICC, 0.41-0.62). Tricuspid regurgitation was noted in 80% and mitral regurgitation in 50% to 60% of patients. Measurements of CO and SV were moderately strong in correlation and agreement which may suggest PAC measurements overestimate actual values. The significance of tricuspid regurgitation and mitral regurgitation during early resuscitation is unknown.


Surgically treated primary malignant tumor of small bowel: a clinical analysis.
To evaluate the clinical presentation, treatment and survival of patients with primary malignant tumor of small bowel (PMTSB). Clinicopathologic data about 141 surgically treated PMTSB patients (91 males and 50 females) at the median age of 53.5 years (range 23-79 years) were retrospectively analyzed. The most common initial clinical features of the patients were intermittent abdominal discomfort or vague abdominal pain (67.4%), abdominal mass (31.2%), bowel obstruction (24.1%), hemotochezia (21.3%), jaundice (16.3%), fever (14.2%), coexistence of bowel perforation and peritonitis (5.7%), coexistence of gastrointestinal bleeding and shock (5.0%), and intraabdominal bleeding (1.4%). Ileum was the most common site of tumor (44.7%), followed by jejunum (30.5%) and duodenum (24.8%). PMTSB had a nonspecific clinical presentation. Segmental bowel resection (n = 81) was the most common surgical procedure, followed by right hemi-colectomy (n = 15), pancreaticoduodenectomy (n = 10), and others (n = 19). Twenty-seven adenocarcinoma patients and 13 malignant lymphoma patients received adjuvant chemotherapy with 5-fluorouracil and cyclophosphamide, adriamycin, vincristine and prednisone, respectively. Information about 120 patients was obtained during the follow-up. The median survival time of PMTSB patients was 20.3 mo. The 1-, 3- and 5-year survival rate was 75.0% (90/120), 40.0% (48/120) and 20.8% (25/120), respectively. Adenocarcinoma was found in 73.7% (42/57), 21.1% (12/57) and 15.8% (9/57) of the patients, respectively. Gastrointestinal stromal tumor was observed in 80.0% (20/25), 72.0% (18/25) and 36.0% (9/25) of the patients, respectively. Carcinoid was detected in 100.0% (15/15), 80.0% (12/15) and 46.7% (7/15) of the patients, respectively. Malignant lymphoma was demonstrated in 69.2% (9/13), 30.8% (4/13) and 0% (0/13) of the patients, respectively. En bloc resection is the principal therapy for most PMTSB and chemotherapy is the important treatment modality for malignant lymphoma and other malignant tumors of small bowel which cannot be radically removed.


Antibody decoration of neurofilaments.
We have decorated neurofilaments with antibodies against three polypeptides (designated here as H [mol wt = 195,000], 45[mol wt = 145,000], and 46[mol wt = 73,000]) in an effort to understand the arrangement of these polypeptides within neurofilaments. The three polypeptides were purified and antibodies were raised against each. The cross-reactivity of the antibodies suggested that each polypeptide contains both shared and unique antigenic determinants. The differential reactivities of each antibody preparation were enhanced by adsorption with the two heterologous polypeptides, and the resulting preparations were used to decorate purified neurofilaments, which were then negatively stained and examined in an electron microscope. The appearance of the antibody-decorated structures led to the following conclusions: All three polypeptides are physically associated with the same neurofilament. However, the disposition of H and 46 within a filament is different; 46 antigens appear to be associated with a "central core" of the filament, whereas H antigens compose a structure more loosely and peripherally attached to the central core and periodically arranged along its axis. The antibody-decorated H-containing structure assumes variable configurations; in some cases it appears asa bridge connecting two filaments; in other cases it appears as a helix wrapping the central core with a period of approximately 1,000 A and an apparent unit length of approximately 1.5 periods. These configurations suggest several functional implications, including the possibility that H is a component of the cross-bridges observed between filaments in situ. We also note that the central core-helix relationship could be used in the design of an intracellular transport motor.


Critical Care Nurses' Experiences During the COVID-19 Pandemic: A US National Survey.
Given critical care nurses' high prepandemic levels of moral distress and burnout, the COVID-19 pandemic will most likely have a tremendous influence on intensive care unit (ICU) nurses' mental health and continuation in the ICU workforce. To describe the experiences of ICU nurses during the COVID-19 pandemic in the United States. Nurses who worked in ICUs in the United States during the COVID-19 pandemic were recruited to complete a survey from October 2020 through early January 2021 through social media and the American Association of Critical-Care Nurses. Three open-ended questions focused on the experiences of ICU nurses during the pandemic. Of 498 nurses who completed the survey, 285 answered the open-ended questions. Nurses reported stress related to a lack of evidence-based treatment, poor patient prognosis, and lack of family presence in the ICU. Nurses perceived inadequate leadership support and inequity within the health care team. Lack of consistent community support to slow the spread of COVID-19 or recognition that COVID-19 was real increased nurses' feelings of isolation. Nurses reported physical and emotional symptoms including exhaustion, anxiety, sleeplessness, and moral distress. Fear of contracting COVID-19 or of infecting family and friends was also prevalent. Intensive care unit nurses in the United States experienced unprecedented and immense burden during the COVID-19 pandemic. Understanding these experiences provides insights into areas that must be addressed to build and sustain an ICU nurse workforce. Studies are needed to further describe nurses' experiences during the COVID-19 pandemic and identify effective resources that support ICU nurse well-being.


Legalizing Cannabis: A physician's primer on the pulmonary effects of marijuana.
Habitual smoking of marijuana is associated with multiple respiratory symptoms such as cough, sputum production, and wheezing .These symptoms are not significantly different from those exhibited by tobacco smokers. Furthermore, endobronchial biopsies of habitual smokers of marijuana and /or tobacco have shown that both marijuana and cigarette smoking cause significant bronchial mucosal histopathology and that these effects are additive. Although marijuana smokers have minimal changes in pulmonary function studies as compared to tobacco smokers, they may develop bullous disease and spontaneous pneumothoraces. The relationship between marijuana smoking and lung cancer remains unclear due to design limitations of the studies published so far. These findings should warn individuals that marijuana smoking may result in serious short-term and long-term respiratory complications, and habitual marijuana use should be viewed with caution. The medical literature so far does not support routine evaluation by pulmonary function tests or imaging studies; until more definitive data is available, we do not recommend the regular use of these tests in the evaluation of habitual marijuana smokers.


Enhanced Killing and Antibiofilm Activity of Encapsulated Cinnamaldehyde against Candida albicans.
Candida sp. impelled opportunistic infection in immune-compromised patients ensuing from asymptomatic colonization to pathogenic forms. Moreover, slow spread of Candida species inducing refractory mucosal and invasive infections brings acute resistance to antifungal drugs. Hence, here we probed the effect of encapsulated preparation of cinnamaldehyde (CNMA) in multilamellar liposomes (ML) against Candida albicans. The efficacy of ML-CNMA against Candida biofilm was assessed by scanning electron microscopy, transmission electron microscopy, as well as light microscopy and its percent inhibition, was determined by XTT [2,3-bis-(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide] and crystal violet assay. ML-CNMA showed more fungicidal activity than free CNMA as well as multilamellar liposomal amphotericin B (ML-Amp B), which was further confirmed by spot test assay and Log-logistic dose-response analysis. Antifungal activity was driven by reactive oxygen species and cellular damage by sustained release of CNMA. Effect on hyphal formation during 48 h in presence/absence of ML-CNMA was observed under a microscope and further substantiated by RT-PCR by amplifying HWP1, the gene responsible for hyphal wall protein formation. Apoptotic programmed cell death was analyzed by FACS analysis which was further confirmed by cytochrome C release assay. This study elucidates the mechanistic insight of the enhanced antifungal activity of ML preparation of CNMA against Candida infections.


Correlation of inhibition of platelet aggregation after clopidogrel with post discharge bleeding events: assessment by different bleeding classifications.
To correlate inhibition of platelet aggregation (IPA) with bleeding events assessed by TIMI, GUSTO, and BleedScore scales in a large cohort of patients with coronary artery disease (CAD) and ischaemic stroke (IS) treated with chronic low-dose aspirin plus clopidogrel. Data from recent trials and registries suggest a link between increased risk of bleeding and cardiovascular mortality. However, the potential association of bleeding risk and IPA is not established. It may play a critical role for the safety of more aggressive platelet inhibition or/and individual tailoring of antiplatelet strategies. Secondary post hoc analyses of 5 microM ADP-induced IPA and bleeding complications assessed by TIMI, GUSTO, and BleedScore scales in a combined data set consisting of patients with documented CAD (n = 246) and previous IS (n = 117). Demographic characteristics differ substantially depending on the underlying vascular disease; however, IPA and bleeding risks were similar between CAD and IS. All three bleeding scales adequately captured serious haemorrhagic events, where the TIMI scale was the most exclusive, whereas BleedScore was the most inclusive. Over half of all patients experienced superficial event(s), most commonly occurring during two to three distinct bleeding episodes. There was no correlation between IPA and duration of antiplatelet therapy. Inhibition of platelet aggregation >50% strongly correlates with minor (r(2) = 0.58, P < 0.001; c-statistic = 0.92), but not severe (r(2) = 0.11, P = 0.038; c-statistic = 0.57), bleeding events. Chronic oral combination antiplatelet regimens are associated with a very high (56.5-60.7%) prevalence of superficial bleeding episodes, which are grossly underestimated in trials and registries. The role of such frequent mild complications for the overall benefit of antiplatelet therapy is entirely unknown, as is their effect on compliance. Although IPA is well suited for defining the risk of minor complications, prediction of more severe bleeding events may be challenging.


[Free radicals and antioxidants: human physiology, pathology and therapeutic aspects].
Oxygen has invaded progressively, and through the ages, an initially anaerobic world. Living organisms had to invent, in the course of evolution, diverse and ingenious defence systems, to survive the toxicity of this element, which was new for them. Strengthened by this experience over billions of years, the present superior organisms, and particularly human species, are thoroughly adapted to 21 per cent of atmospheric oxygen. Nevertheless, the equilibrium is fragile and the menace of oxygen hovers continually. This deleterious potential of oxygen is attributed to the formation, in vivo, of free radicals, a free radical being, by definition, any chemical species possessing one or several mismatched electrons. These free radicals are, in general, very active. They trigger chain reactions able to damage the different constituents of the living organism. Basic oxygen, must be pre-activated to manifest its toxicity. Such an activation can be achieved in two ways: it can be photodynamic, ending mainly in singlet oxygen, it can be reducing, followed by the formation of the anion hydrogen peroxide and of radical hydroxyl; the latter is the most reactive chemical species in the biological world. The reductive process is accelerated in the presence of transition metals, such as iron and copper, and/or specific enzymes (monoxygenase and certain oxydases). This activation takes place in different cellular compartments: mitochondria, microsomes, peroxysomes, cytoplasmic membrane. To this potential toxicity of oxygen the organism opposes different anti-oxidant defence systems. A first group works up the radical chain, inhibiting activation mechanisms. Such a group, as a consequence, warns of the initiation of radical reactions. The second group neutralizes the free radicals already formed and thus stops the chain of propagation. In this group can be found detoxifying enzymes, notably superoxide dismutase and catalase, producing jointly peroxidases, particularly peroxidase glutathions. Such enzymes for the most part have trace elements as cofactors. In this second group can also be found various molecules which act like 'substrate suicide', or as an anti-oxidant shield. Among these molecules, some can act in the lipidic phase, such as tocopherols, carotenoïds and ubiquinones. Other molecules which are lipophobic, mainly ascorbic acid and uric acid, are active in a hydrated environment. In the case of a weakening of such an antioxidant defence or excess production of radicals, a state of oxidative stress occurs. Uncontrolled, these radicals will damage different biological targets: lipids, DNA, proteins. Disturbances of cellular metabolism will occur, unless corrective defences intervene. The identification of these radical phenomena is an obligatory stage. But because of the very short life span of free radicals, identification poses a real analytical problem. However, three approaches are possible: identification of free radicals, either directly by means of paramagnetic electron resonance, or indirectly by identifying some more stable intermediates. evaluation of the traces of radical attack on biological molecules, for example by high performance liquid chromatography, gas-liquid chromatography, colorimetric tests, estimation of the antioxidant status, for example by colorimetric tests, immunoenzymatic methods, high performance liquid chromatography.


p18Ink4c and p53 Act as tumor suppressors in cyclin D1-driven primitive neuroectodermal tumor.
The retinoblastoma (RB) tumor suppressor pathway is likely important in primitive neuroectodermal tumors (PNET) of the brain. In fact, 10% to 15% of children born with RB mutations develop brain PNETs, commonly in the pineal gland. Cyclin D1, which in association with cyclin-dependent kinase (Cdk) 4 and Cdk6 phosphorylates and inactivates the RB protein, is expressed in 40% of sporadic medulloblastoma, a PNET of the cerebellum. To understand tumorigenic events cooperating with RB pathway disruption in brain PNET, we generated a transgenic mouse where cyclin D1 was expressed in pineal cells. Cyclin D1 enhanced pinealocyte proliferation, causing pineal gland enlargement. However, proliferation ceased beyond 2 weeks of age with reversal of Cdk4-mediated Rb phosphorylation despite continued expression of the transgene, and the pineal cells showed heterochromatin foci suggestive of a senescent-like state. In the absence of the p53 tumor suppressor, cell proliferation continued, resulting in pineal PNET that limited mouse survival to approximately 4 months. Interestingly, the Cdk inhibitor p18(Ink4c) was induced in the transgenic pineal glands independently of p53, and transgenic mice that lacked Ink4c developed invasive PNET, although at an older age than those lacking p53. Analogous to our mouse model, we found that children with heritable RB often had asymptomatic pineal gland enlargement that only rarely progressed to PNET. Our finding that the Cdk4 inhibitor p18(Ink4c) is a tumor suppressor in cyclin D1-driven PNET suggests that pharmacologic interventions to inhibit Cdk4 activity may be a useful chemoprevention or therapeutic strategy in cancer driven by primary RB pathway disruption.


Predictive Validity of a 2-Question Alcohol Screen at 1-, 2-, and 3-Year Follow-up.
The National Institute on Alcohol Abuse and Alcoholism (NIAAA) 2-question screen is a valid adolescent alcohol screening tool. No studies have examined if this tool predicts future alcohol problems. We conducted a study at 16 pediatric emergency departments to determine the tool's predictive validity for alcohol misuse and alcohol use disorders (AUDs). Participants (N = 4834) completed a baseline assessment battery. A subsample of participants completed the battery at 1, 2, and 3 years follow up. Of the 2209 participants assigned to follow-up, 1611 (73%) completed a 1-year follow-up, 1591 (72%) completed a 2-year follow-up, and 1377 (62%) completed a 3-year follow-up. The differences in AUDs between baseline NIAAA screen nondrinkers and lower-risk drinkers were statistically significant at 1 year (P = .0002), 2 years (P <.0001), and 3 years (P = .0005), as were the differences between moderate- and highest-risk drinkers at 1 and 2 years (P < .0001 and P = .0088, respectively) but not at 3 years (P = .0758). The best combined score for sensitivity (86.2% at 1 year, 75.6% at 2 years, and 60.0% at 3 years) and specificity (78.1% at 1 year, 79.2% at 2 years, and 80.0% at 3 years) was achieved by using "lower risk" and higher as a cutoff for the prediction of a Diagnostic and Statistical Manual of Mental Disorders, Fifth Edition diagnosis. The NIAAA 2-question screen can accurately characterize adolescent risk for future AUDs. Future studies are needed to determine optimaluse of the screen.


A concept for NASA's Mars 2016 astrobiology field laboratory.
The Mars Program Plan includes an integrated and coordinated set of future candidate missions and investigations that meet fundamental science objectives of NASA and the Mars Exploration Program (MEP). At the time this paper was written, these possible future missions are planned in a manner consistent with a projected budget profile for the Mars Program in the next decade (2007-2016). As with all future missions, the funding profile depends on a number of factors that include the exact cost of each mission as well as potential changes to the overall NASA budget. In the current version of the Mars Program Plan, the Astrobiology Field Laboratory (AFL) exists as a candidate project to determine whether there were (or are) habitable zones and life, and how the development of these zones may be related to the overall evolution of the planet. The AFL concept is a surface exploration mission equipped with a major in situ laboratory capable of making significant advancements toward the Mars Program's life-related scientific goals and the overarching Vision for Space Exploration. We have developed several concepts for the AFL that fit within known budget and engineering constraints projected for the 2016 and 2018 Mars mission launch opportunities. The AFL mission architecture proposed here assumes maximum heritage from the 2009 Mars Science Laboratory (MSL). Candidate payload elements for this concept were identified from a set of recommendations put forth by the Astrobiology Field Laboratory Science Steering Group (AFL SSG) in 2004, for the express purpose of identifying overall rover mass and power requirements for such a mission. The conceptual payload includes a Precision Sample Handling and Processing System that would replace and augment the functionality and capabilities provided by the Sample Acquisition Sample Processing and Handling system that is currently part of the 2009 MSL platform.


Improving the linearity of infrared diffuse reflection spectroscopy data for quantitative analysis: an application in quantifying organophosphorus contamination in soil.
Diffuse reflection data are presented for ethyl methylphosphonate in a fine Utah dirt sample as a model system for organophosphate-contaminated soil. The data revealed a chemometric artifact when the spectra were represented in Kubelka-Munk units that manifests as a linear dependence of spectral peak height on variations in the observed baseline position (i.e., the position of the observed transmission intensity where no absorption features occur in the sample spectrum). We believe that this artifact is the result of the mathematical process by which the raw data are converted into Kubelka-Munk units, and we developed a numerical strategy for compensating for the observed effect and restoring chemometric precision to the diffuse reflection data for quantitative analysis while retaining the benefits of linear calibration afforded by the Kubelka-Munk approach. We validated our Kubelka-Munk correction strategy by repeating the experiment using a simpler system--pure caffeine in potassium bromide. The numerical preprocessing includes conventional multiplicative scatter correction coupled with a baseline offset correction that facilitates the use of quantitative diffuse reflection data in the Kubelka-Munk formalism for the quantitation of contaminants in a complex soil matrix, but is also applicable to more fundamental diffuse reflection quantitative analysis experiments.


Worry and rumination: Explanatory roles in the relation between pain and anxiety and depressive symptoms among college students with pain.
Pain affects a significant proportion of college students in the United States and has been linked to anxiety and depressive symptoms. Rumination and worry, two transdiagnostic factors linked to comorbidity, may explain the relationship between pain and mental health symptoms. Current Study: The current study examined worry and rumination as explanatory factors in the relationship between pain and anxiety and depressive symptoms in a sample of college students with pain (n = 1,577; 79.9% female). Results indicated that both rumination and worry explained the relationship between pain and depressive and social anxiety symptoms, while rumination alone explained the relationship between pain and anxious arousal symptoms. The current study provides novel empirical evidence that worry and rumination each help explain the relationship between pain and anxiety and depressive symptoms among college students with current pain, and college students in pain may benefit from targeted psychosocial strategies aimed at decreasing worry and ruminative responses.


Mouse models of Tay-Sachs and Sandhoff diseases differ in neurologic phenotype and ganglioside metabolism.
Tay-Sachs and Sandhoff diseases are clinically similar neurodegenerative disorders. These two sphingolipidoses are characterized by a heritable absence of beta-hexosaminidase A resulting in defective GM2 ganglioside degradation. Through disruption of the Hexa and Hexb genes in embryonic stem cells, we have established mouse models corresponding to each disease. Unlike the two human disorders, the two mouse models show very different neurologic phenotypes. Although exhibiting biochemical and pathologic features of the disease, the Tay-Sachs model showed no neurological abnormalities. In contrast, the Sandhoff model was severely affected. The phenotypic difference between the two mouse models is the result of differences in the ganglioside degradation pathway between mice and humans.


The hidden research paper.
To determine whether the views expressed in a research paper are accurate representations of contributors' opinions about the research being reported. Purposive sampling of 10 research articles published in The Lancet; qualitative analysis of answers to 6 questions about the meaning of the study put to contributors who were listed on the byline of these articles. Fifty-four contributors listed on the bylines of the 10 articles were evaluated, and answers to questions were compared between contributors within research groups and against the published research report. A total of 36 (67%) of 54 contributors replied to this survey. Important weaknesses were often admitted on direct questioning but were not included in the published article. Contributors frequently disagreed about the importance of their findings, implications, and directions for future research. I could find no effort to study systematically past evidence relating to the investigators' own findings in either survey responses or the published article. Overall, the diversity of contributor opinion was commonly excluded from the published report. I found that discussion sections were haphazardly organized and did not deal systematically with important questions about the study. A research paper rarely represents the opinions of those scientists whose work it reports. The findings described herein reveal evidence of (self-)censored criticism, obscured meanings, confused assessment of implications, and failures to indicate directions for future research. There is now empirical support for the introduction of structured discussion sections in research papers. Editors might also explore ways to recover the plurality of contributors' opinions.


NEDD8 recruits E2-ubiquitin to SCF E3 ligase.
NEDD8/Rub1 is a ubiquitin (Ub)-like post-translational modifier that is covalently linked to cullin (Cul)-family proteins in a manner analogous to ubiquitylation. NEDD8 is known to enhance the ubiquitylating activity of the SCF complex (composed of Skp1, Cul-1, ROC1 and F-box protein), but the mechanistic role is largely unknown. Using an in vitro reconstituted system, we report here that NEDD8 modification of Cul-1 enhances recruitment of Ub-conjugating enzyme Ubc4 (E2) to the SCF complex (E3). This recruitment requires thioester linkage of Ub to Ubc4. Our findings indicate that the NEDD8-modifying system accelerates the formation of the E2-E3 complex, which stimulates protein polyubiquitylation.


Acute Limonene Toxicity in Escherichia coli Is Caused by Limonene Hydroperoxide and Alleviated by a Point Mutation in Alkyl Hydroperoxidase AhpC.
Limonene, a major component of citrus peel oil, has a number of applications related to microbiology. The antimicrobial properties of limonene make it a popular disinfectant and food preservative, while its potential as a biofuel component has made it the target of renewable production efforts through microbial metabolic engineering. For both applications, an understanding of microbial sensitivity or tolerance to limonene is crucial, but the mechanism of limonene toxicity remains enigmatic. In this study, we characterized a limonene-tolerant strain of Escherichia coli and found a mutation in ahpC, encoding alkyl hydroperoxidase, which alleviated limonene toxicity. We show that the acute toxicity previously attributed to limonene is largely due to the common oxidation product limonene hydroperoxide, which forms spontaneously in aerobic environments. The mutant AhpC protein with an L-to-Q change at position 177 (AhpC(L177Q)) was able to alleviate this toxicity by reducing the hydroperoxide to a more benign compound. We show that the degree of limonene toxicity is a function of its oxidation level and that nonoxidized limonene has relatively little toxicity to wild-type E. coli cells. Our results have implications for both the renewable production of limonene and the applications of limonene as an antimicrobial.


Comparison of efficacy of thoracic paravertebral block with oblique subcostal transversus abdominis plane block in open cholecystectomy.
Sensory afferent nerve branches of lower six thoracic and upper lumbar nerves innervate the anterior abdominal wall and are the therapeutic focus of local anesthetics to provide analgesia for the abdominal surgical incision. Central neuraxial and regional analgesia can provide better control of pain due to right subcostal incision used in open cholecystectomy and attenuate the need for opioids. The earlier studies which showed the benefit of the thoracic paravertebral block (TPVB) for analgesia after upper abdominal surgeries did not compare TPVB with oblique subcostal transversus abdominis plane (OSTAP) block. Therefore, the current study compares the analgesic efficacy of TPVB and OSTAP block in open cholecystectomy. Seventy consenting adults scheduled for open cholecystectomy were allocated to one of the two groups: ultrasound-guided TPVB (Group I) and ultrasound-guided OSTAP block (Group II). The primary objective of this study is to assess and compare tramadol consumption in 48 h in both the groups along with VAS in post anesthesia care unit, and then at 2, 4, 6, 8, 12, 24, and 48 h. The secondary objective of the study is to assess the incidence of PONV. The number of doses of rescue analgesia required was less in Group I when compared with Group II (P < 0.001). Patients in Group I had significantly lower pain scores than Group II. Although in the initial 8 h, both groups had comparable pain scores, after 8 h, these scores were significantly lower in patients in Group I. Less postoperative nausea and vomiting was seen in Group I patients (11.7%) in comparison to Group II (18.1%). Ultrasound-guided TPVB is superior to OSTAP block because of its association with decreased postoperative opioid consumption, lower VAS score, and reduction in opioid-related side effects. Thus, it should be strongly considered as a part of multimodal analgesia regimen in upper abdominal surgeries.


The role of social cognition skills and social determinants of health in predicting symptoms of mental illness.
Social factors, such as social cognition skills (SCS) and social determinants of health (SDH), may be vital for mental health, even when compared with classical psycho-physical predictors (demographic, physical, psychiatric, and cognitive factors). Although major risk factors for psychiatric disorders have been previously assessed, the relative weight of SCS and SDH in relation to classical psycho-physical predictors in predicting symptoms of mental disorders remains largely unknown. In this study, we implemented multiple structural equation models (SEM) from a randomized sample assessed in the Colombian National Mental Health Survey of 2015 (CNMHS, n = 2947, females: 1348) to evaluate the role of SCS, SDH, and psycho-physical factors (totaling 17 variables) as predictors of mental illness symptoms (anxiety, depression, and other psychiatric symptoms). Specifically, we assessed the structural equation modeling of (a) SCS (emotion recognition and empathy skills); (b) SDH (including the experience of social adversities and social protective factors); (c) and classical psycho-physical factors, including psychiatric antecedents, physical-somatic factors (chronic diseases), and cognitive factors (executive functioning). Results revealed that the emotion recognition skills, social adverse factors, antecedents of psychiatric disorders and chronic diseases, and cognitive functioning were the best predictors of symptoms of mental illness. Moreover, SCS, particularly emotion recognition skills, and SDH (experiences of social adversities, familial, and social support networks) reached higher predictive values of symptoms than classical psycho-physical factors. Our study provides unprecedented evidence on the impact of social factors in predicting symptoms of mental illness and highlights the relevance of these factors to track early states of disease.


Gut microbiota, chemotherapy and the host: the influence of the gut microbiota on cancer treatment.
The gut microbiota exists in a dynamic balance between symbiosis and pathogenesis and can influence almost any aspect of host physiology. Growing evidence suggests that the gut microbiota not only plays a key role in carcinogenesis but also influences the efficacy and toxicity of anticancer therapy. The microbiota modulates the host response to chemotherapy via numerous mechanisms, including immunomodulation, xenometabolism and alteration of community structure. Furthermore, exploitation of the microbiota offers opportunities for the personalisation of chemotherapeutic regimens and the development of novel therapies. In this article, we explore the host-chemotherapeutic microbiota axis, from basic science to clinical research, and describe how it may change the face of cancer treatment.


Video-based assessment for laparoscopic fundoplication: initial development of a robust tool for operative performance assessment.
While better technical performance correlates with improved outcomes, there is a lack of procedure-specific tools to perform video-based assessment (VBA). SAGES is developing a series of VBA tools with enough validity evidence to allow reliable measurement of surgeon competence. A task force was established to develop a VBA tool for laparoscopic fundoplication using an evidence-based process that can be replicated for additional procedures. The first step in this process was to seek content validity evidence. Forty-two subject matter experts (SME) in laparoscopic fundoplication were interviewed to obtain consensus on procedural steps, identify potential variations in technique, and to generate an inventory of required skills and common errors. The results of these interviews were used to inform creation of a task inventory questionnaire (TIQ) that was delivered to a larger SME group (n = 188) to quantify the criticality and difficulty of the procedural steps, the impact of potential errors associated with each step, the technical skills required to complete the procedure, and the likelihood that future techniques or technologies may change the presence or importance of any of these factors. Results of the TIQ were used to generate a list of steps, skills, and errors with strong validity evidence. Initial SMEs interviewed included fellowship program directors (45%), recent fellows (24%), international surgeons (19%), and highly experienced super SMEs with quality outcomes data (12%). Qualitative analysis of interview data identified 6 main procedural steps (visualization, hiatal dissection, fundus mobilization, esophageal mobilization, hiatal repair, and wrap creation) each with 2-5 sub steps. Additionally, the TIQ identified 5-10 potential errors for each step and 11 key technical skills required to perform the procedure. Based on the TIQ, the mean criticality and difficulty scores for the 11/21 sub steps included in the final scoring rubric is 4.66/5 (5 = absolutely essential for patient outcomes) and 3.53/5 (5 = difficulty level requires significant experience and use of alternative strategies to accomplish consistently), respectively. The mean criticality and frequency scores for the 9/11 technical skills included is 4.51/5 and 4.51/5 (5 = constantly used ≥ 80% of the time), respectively. The mean impact score of the 42/47 errors incorporated into the final rubric is 3.85/5 (5 = significant error that is unrecoverable, or even if recovered, likely to have a negative impact on patient outcome). A rigorous, multi-method process has documented the content validity evidence for the SAGES video-based assessment tool for laparoscopic fundoplication. Work is ongoing to pilot the assessment tool on recorded fundoplication procedures to establish reliability and further validity evidence.


Novel diagnostic and therapeutic techniques for surveillance of dysplasia in patients with inflammatory bowel disease.
The risk for developing dysplasia and colorectal cancer in patients with longstanding inflammatory bowel disease (IBD) involving the colon is well documented. Random biopsies during white-light, standard-definition colonoscopy (33 to 50 biopsies) with or without dye spraying chromoendoscopy has been the recommended strategy in North America to detect dysplastic lesions in IBD. However, there are several limitations to this approach including poor physician adherence, poor sensitivity, increased procedure time and considerable cost. The new generation of high-definition endoscopes with electronic filter technology provide an opportunity to visualize colonic mucosal and vascular patterns in minute detail, and to identify subtle flat, multifocal, polypoid and pseudopolypoid neoplastic and non-neoplastic lesions. The application of these new technologies in IBD is slowly being adopted in clinical practice. In addition, the advent of confocal laser endomicroscopy provides an opportunity to explore real-time histology, thus redefining the understanding and characterization of the lesions in IBD. There is emerging evidence that serrated adenomas are also associated with longstanding IBD colitis and may be recognized as another important contributing factor to colorectal cancer development. The circumscribed neoplastic lesions can be treated using endoscopic therapeutic management such as mucosal resection or, especially, endoscopic submucosal dissection. This may replace panproctocolectomy in selected patients. The authors review the potential of these techniques to transform endoscopic diagnosis and therapeutic management of dysplasia in IBD.


Sialylated IgG-Fc: a novel biomarker of chronic inflammatory demyelinating polyneuropathy.
Sialylation in Fc portion of IgG plays a crucial role in the pathogenesis of autoimmune diseases and the working mechanism of intravenous immunoglobulin (IVIG). We aim to test whether IgG-Fc sialylation is a biomarker of disease activity for chronic inflammatory demyelinating polyneuropathy (CIDP). By using specific lectins for sialylation, galactosylation and agalactosylation, lectin-enzyme assay and lectin blotting with pretreatment of IgG degradating enzyme of Streptococcus pyogenes were performed to compare the glycosylation levels of serum IgG-Fc (1) between patients of untreated CIDP (n=107) and normal control subjects (n=27), (2) among patients with untreated CIDP of different clinical severities and (3) before and after IVIG treatment of patients with CIDP (n=12). Sialylation and galactosylation of IgG-Fc were significantly reduced in patients with CIDP than normal control subjects (p=0.003 and 0.033, respectively), whereas agalactosylation was increased in CIDP (p=0.21). Ratios of sialylated/agalactosylated IgG-Fc levels were significantly reduced in CIDP (p<0.001) and inversely related to disease severity (p=0.044). After IVIG treatment, levels of sialylated IgG-Fc significantly increased (p=0.003). Sialylation of IgG-Fc is reduced in CIDP. Its level correlated with clinical severity and increased after IVIG treatment. Sialylated as well as ratio of sialylated/agalactosylated IgG-Fc could be new measures to monitor the disease severity and treatment status in CIDP.


Minim typing--a rapid and low cost MLST based typing tool for Klebsiella pneumoniae.
Here we report a single nucleotide polymorphism (SNP) based genotyping method for Klebsiella pneumoniae utilising high-resolution melting (HRM) analysis of fragments within the multilocus sequence typing (MLST) loci. The approach is termed mini-MLST or Minim typing and it has previously been applied to Streptococcus pyogenes, Staphylococcus aureus and Enterococcus faecium. Six SNPs were derived from concatenated MLST sequences on the basis of maximisation of the Simpsons Index of Diversity (D). DNA fragments incorporating these SNPs and predicted to be suitable for HRM analysis were designed. Using the assumption that HRM alleles are defined by G+C content, Minim typing using six fragments was predicted to provide a D = 0.979 against known STs. The method was tested against 202 K. pneumoniae using a blinded approach in which the MLST analyses were performed after the HRM analyses. The HRM-based alleles were indeed in accordance with G+C content, and the Minim typing identified known STs and flagged new STs. The tonB MLST locus was determined to be very diverse, and the two Minim fragments located herein contribute greatly to the resolving power. However these fragments are refractory to amplification in a minority of isolates. Therefore, we assessed the performance of two additional formats: one using only the four fragments located outside the tonB gene (D = 0.929), and the other using HRM data from these four fragments in conjunction with sequencing of the tonB MLST fragment (D = 0.995). The HRM assays were developed on the Rotorgene 6000, and the method was shown to also be robust on the LightCycler 480, allowing a 384-well high through-put format. The assay provides rapid, robust and low-cost typing with fully portable results that can directly be related to current MLST data. Minim typing in combination with molecular screening for antibiotic resistance markers can be a powerful surveillance tool kit.


An evolutionary perspective on salt, hypertension, and human genetic variability.
Natural selection for electrolyte conservation has likely been the norm throughout human evolutionary history. However, the current patterns of excessive dietary salt intake create the potential for salt overload. Under these conditions, hypertension may be considered an expected pathological response to an evolutionarily new constraint. The transatlantic Middle Passage may have created a genetic bottleneck for salt conservation in African-Americans. Although the initial consequences of this important historical event probably constricted genetic variability and further magnified the potential for salt-sensitive hypertension, the Middle Passage undoubtedly also served as a more generalized major source of environmental stress and may have stimulated subsequent hereditary diversity in the survivors of this holocaust and their descendants. Accelerated rates of mutation, genetic recombination, and transposable genetic elements in conjunction with enhanced opportunities for gene flow, new selective pressures, and drift have all contributed to the tremendous heterogeneity of contemporary African-Americans. It is unlikely that a single genetic event, even of the severity of the Middle Passage, can account comprehensively for the apparent susceptibility of this macroethnic group to high blood pressure and hypertension.


Nanogels and Microgels: From Model Colloids to Applications, Recent Developments, and Future Trends.
Nanogels and microgels are soft, deformable, and penetrable objects with an internal gel-like structure that is swollen by the dispersing solvent. Their softness and the potential to respond to external stimuli like temperature, pressure, pH, ionic strength, and different analytes make them interesting as soft model systems in fundamental research as well as for a broad range of applications, in particular in the field of biological applications. Recent tremendous developments in their synthesis open access to systems with complex architectures and compositions allowing for tailoring microgels with specific properties. At the same time state-of-the-art theoretical and simulation approaches offer deeper understanding of the behavior and structure of nano- and microgels under external influences and confinement at interfaces or at high volume fractions. Developments in the experimental analysis of nano- and microgels have become particularly important for structural investigations covering a broad range of length scales relevant to the internal structure, the overall size and shape, and interparticle interactions in concentrated samples. Here we provide an overview of the state-of-the-art, recent developments as well as emerging trends in the field of nano- and microgels. The following aspects build the focus of our discussion: tailoring (multi)functionality through synthesis; the role in biological and biomedical applications; the structure and properties as a model system, e.g., for densely packed arrangements in bulk and at interfaces; as well as the theory and computer simulation.


Predictors of long-term treatment effect of rivastigmine in Alzheimer's disease: a role for beta-amyloid plasma levels?
Cholinesterase inhibitors (ChEI) are currently the mainstream symptomatic treatment of patients with Alzheimer's disease (AD). To this end, the response to the treatment with ChEI is clinically difficult to predict. Several demographic, clinical and biological variables have been proposed as pre-treatment predictors of long-term therapy efficacy. The aim of the study was to confirm our initial observations of the significance of a change in plasma levels of beta-amyloid (Abeta) peptides after initial treat-ment with rivastigmine for predicting clinical response to ChEI. Fifty-four carefully selected subjects (37 females) satisfying criteria for mild (n = 25) or moderate (n = 29) AD were included in the study. Rivastigmine was prescribed at the initial dose of 3 mg/day b.i.d.; the dose was escalated to the maximum tolerated one in at least 4-week intervals. The response to treatment was assessed using the ADAS-Cog and CDR scales. Whole blood samples were collected twice: before the first rivastigmine dose and at the 2nd week on active treatment. Levels of Abeta(1-40) and Abeta(1-42) were measured in plasma using a commercially available ELISA. We confirmed that higher initial disease severity (higher ADAS-Cog scores) and the increase in the con-centration of plasma Abeta(1-42) peptide following 2 weeks of treatment with an initial dose of rivastigmine increased the chance of a clinically meaningful response to ChEI therapy in AD patients after 2 years of follow-up. A change in plasma Abeta(1-42) level might constitute a novel biochemical predictor of long-term rivastigmine treatment efficacy in AD.


Renal Decapsulation Prevents Intrinsic Renal Compartment Syndrome in Ischemia-Reperfusion-Induced Acute Kidney Injury: A Physiologic Approach.
Acute kidney injury is a serious complication with unacceptably high mortality that lacks of specific curative treatment. Therapies focusing on the hydraulic behavior have shown promising results in preventing structural and functional renal impairment, but the underlying mechanisms remain understudied. Our goal is to assess the effects of renal decapsulation on regional hemodynamics, oxygenation, and perfusion in an ischemic acute kidney injury experimental model. In piglets, intra renal pressure, renal tissue oxygen pressure, and dysoxia markers were measured in an ischemia-reperfusion group with intact kidney, an ischemia-reperfusion group where the kidney capsule was removed, and in a sham group. Decapsulated kidneys displayed an effective reduction of intra renal pressure, an increment of renal tissue oxygen pressure, and a better performance in the regional delivery, consumption, and extraction of oxygen after reperfusion, resulting in a marked attenuation of acute kidney injury progression due to reduced structural damage and improved renal function. Our results strongly suggest that renal decapsulation prevents the onset of an intrinsic renal compartment syndrome after ischemic acute kidney injury.


The efficacy of ginkgo for elderly people with dementia and age-associated memory impairment: new results of a randomized clinical trial.
To evaluate the efficacy, the dose-dependence, and the durability of the effect of the ginkgo biloba special extract EGb 761 (ginkgo) in older people with dementia or age-associated memory impairment. A 24-week, randomized, double-blind, placebo-controlled, parallel-group, multicenter trial. Homes for the elderly in the southern part of the Netherlands. Older persons with dementia (either Alzheimer's dementia or vascular dementia; mild to moderate degree) or age-associated memory impairment (AAMI). 214 Participants were recruited from 39 homes for the elderly. The participants were allocated randomly to treatment with EGb 761 (2 tablets per day, total dosage either 240 (high dose) or 160 (usual dose) mg/day) or placebo (0 mg/d). The total intervention period was 24 weeks. After 12 weeks of treatment, the initial ginkgo users were randomized once again to either continued ginkgo treatment or placebo treatment. Initial placebo use was prolonged after 12 weeks. Outcomes were assessed after 12 and 24 weeks of intervention. Outcome measures included neuropsychological testing (trail-making speed (NAI-ZVT-G), digit memory span (NAI-ZN-G), and verbal learning (NAI-WL)), clinical assessment (presence and severity of geriatric symptoms (SCAG), depressive mood (GDS), self-perceived health and memory status (report marks)), and behavioral assessment (self-reported level of instrumental daily life activities). An intention-to-treat analysis showed no effect on each of the outcome measures for participants who were assigned to ginkgo (n = 79) compared with placebo (n = 44) for the entire 24-week period. After 12 weeks of treatment, the combined high dose and usual dose ginkgo groups (n = 166) performed slightly better with regard to self-reported activities of daily life but slightly worse with regard to self-perceived health status compared with the placebo group (n = 48). No beneficial effects of a higher dose or a prolonged duration of ginkgo treatment were found. We could not detect any subgroup that benefited from ginkgo. Ginkgo use was also not associated with the occurrence of (serious) adverse events. The results of our trial suggest that ginkgo is not effective as a treatment for older people with mild to moderate dementia or age-associated memory impairment. Our results contrast sharply with those of previous ginkgo trials.


Immobilizing a visible light-responsive photocatalyst on a recyclable polymeric composite for floating and suspended applications in water treatment.
A visible light responsive TiO2/Ag3PO4 (10 : 1) nanocomposite was prepared and successfully immobilized (12 wt%) in a spherical polymeric matrix consisting of polysulfone and alginate (10 : 6). The resulted beads featured a sponge-like structure with interconnected macrovoids and micropores, and showed high adsorption and visible-light photocatalytic activity towards various wastewater pollutants, including the widely used dye - methylene blue (k = 0.0321 min-1), and two emerging pharmaceutical contaminants - diclofenac (k = 0.018 min-1) and triclosan (k = 0.052 min-1). As determined, the ˙OH radical and h+ are the primary reactive oxygen species responsible for the photodegradation. The composite photocatalytic beads are also effective in bacterial inactivation and degradation of acyl-homoserine lactones (AHLs), the bacterial quorum sensing autoinducers triggering biofilms, thus exhibiting a promising future in wastewater disinfection and biofilm retardation. Additionally, these beads could be used in inter-switchable suspended or buoyant forms, and be effectively regenerated by H2O2 treatment, and used for multiple cycles without any significant loss in photoactivity. With these unique features, the prepared visible-light photocatalytic beads could be easily applied in large-scale water and wastewater treatment systems.


Practical tests for distinguishing slow light from saturable absorption.
A series of practical tests of slow light (light with reduced group velocity) in saturable absorbers is proposed. These include experimental tests for saturable absorption, which can mimic slow light effects in saturable media, the dependence of slow light on the mutual coherence of pump and probe, since both slow and fast light effects can be simulated with incoherent sources, and the influence of polarization. The principal requirements for practical observation of spectral hole burning are reviewed and shown to be achievable for a wide range of saturable media with the narrow line sources now available.


In vivo volumetric imaging of chicken retina with ultrahigh-resolution spectral domain optical coherence tomography.
The chicken retina is an established animal model for myopia and light-associated growth studies. It has a unique morphology: it is afoveate and avascular; oxygen and nutrition to the inner retina is delivered by a vascular tissue (pecten) that protrudes into the vitreous. Here we present, to the best of our knowledge, the first in vivo, volumetric high-resolution images of the chicken retina. Images were acquired with an ultrahigh-resolution optical coherence tomography (UHROCT) system with 3.5 µm axial resolution in the retina, at the rate of 47,000 A-scans/s. Spatial variations in the thickness of the nerve fiber and ganglion cell layers were mapped by segmenting and measuring the layer thickness with a semi-automatic segmentation algorithm. Volumetric visualization of the morphology and morphometric analysis of the chicken retina could aid significantly studies with chicken retinal models of ophthalmic diseases.


Knowledge and behaviours related to herbal remedies: a cross-sectional epidemiological study in adults in Middle Anatolia, Turkey.
The use of herbal remedies is common in Turkey. This study aimed to define the patterns of herbal remedy use among subjects aged 18 or above and to describe factors associated with use of herbal remedies. A cross-sectional study was conducted with a multistage sampling method between September 2007 and April 2008. A total of 3876 adults participated (98.1% response rate), and 1518 (39.2%) admitted using herbal remedies (95% CI 37.6-40.7%). Univariate analyses showed that compared with non-users, herbal users were mostly female, were more highly educated and were more likely to live in smaller households of one to four people. Logistic regression showed that being aged 18-27 (OR = 1.23, P = 0.028), being female (OR = 2.22, P < 0.001), being educated in a secondary school (OR = 1.47, P < 0.001) or high school (OR = 2.77, P < 0.001), perceiving their health status as 'good' (OR = 2.61, P < 0.001) and having health-related problems (OR = 2.80, P < 0.001) were the factors associated with the use of herbal remedies. The most commonly used herbs were lime, mint, rosehip and lemon. The reasons for using herbal remedies included that they are natural products (79.8% of the 1518 users), for health enhancement (58.9%) and to overcome health problems (32.2%). The decision to use herbal remedies was mainly based on recommendations from the mass media (45.1%). Only 29.1% of users obtained information from their physicians or health providers, and only 37.9% informed their doctors. Nearly three in five people in this study reported using a herbal remedy to overcome health problems or for health enhancement. Herbal remedies are likely to be used by the young people, females, those with higher education, those with good or excellent perceived health status and those with chronic illness, and it seems essential to offer informational programmes for them. The lack of communication on herbal remedies between patient and physician needs to be addressed.


Proteolytic cleavage of chemerin protein is necessary for activation to the active form, Chem157S, which functions as a signaling molecule in glioblastoma.
Chemerin is a chemoattractant involved in innate and adaptive immunity as well as an adipokine implicated in adipocyte differentiation. Chemerin circulates as an inactive precursor in blood whose bioactivity is closely regulated through proteolytic processing at its C terminus. We developed methodology for production of different recombinant chemerin isoforms (chem163S, chem157S, and chem155A) which allowed us to obtain large quantities of these proteins with purity of >95%. Chem158K was generated from chem163S by plasmin cleavage. Characterization by mass spectrometry and Edman degradation demonstrated that both the N and C termini were correct for each isoform. Ca(2+) mobilization assays showed that the EC(50) values for chem163S and chem158K were 54.2 ± 19.9 nm and 65.2 ± 13.2 nm, respectively, whereas chem157S had a ∼50-fold higher potency with an EC(50) of 1.2 ± 0.7 nm. Chem155A had no agonist activity and weak antagonist activity, causing a 50% reduction of chem157S activity at a molar ratio of 100:1. Similar results were obtained in a chemotaxis assay. Because chem158K is the dominant form in cerebrospinal fluid from patients with glioblastoma (GBM), we examined the significance of chemerin in GBM biology. In silico analysis showed chemerin mRNA was significantly increased in tissue from grade III and IV gliomas. Furthermore, U-87 MG cells, a human GBM line, express the chemerin receptors, chemokine-like receptor 1 and chemokine receptor-like 2, and chem157S triggered Ca(2+) flux. This study emphasized the necessity of appropriate C-terminal proteolytic processing to generate the likely physiologic form of active chemerin, chem157S, and suggested a possible role in malignant GBM.


Distribution of selected healthcare resources for influenza pandemic response in Cambodia.
Human influenza infection poses a serious public health threat in Cambodia, a country at risk for the emergence and spread of novel influenza viruses with pandemic potential. Prior pandemics demonstrated the adverse impact of influenza on poor communities in developing countries. Investigation of healthcare resource distribution can inform decisions regarding resource mobilization and investment for pandemic mitigation. A health facility survey performed across Cambodia obtained data on availability of healthcare resources important for pandemic influenza response. Focusing on five key resources considered most necessary for treating severe influenza (inpatient beds, doctors, nurses, oseltamivir, and ventilators), resource distributions were analyzed at the Operational District (OD) and Province levels, refining data analysis from earlier studies. Resources were stratified by respondent type (hospital vs. District Health Office [DHO]). A summary index of distribution inequality was calculated using the Gini coefficient. Indices for local spatial autocorrelation were measured at the OD level using geographical information system (GIS) analysis. Finally, a potential link between socioeconomic status and resource distribution was explored by mapping resource densities against poverty rates. Gini coefficient calculation revealed variable inequality in distribution of the five key resources at the Province and OD levels. A greater percentage of the population resides in areas of relative under-supply (28.5%) than over-supply (21.3%). Areas with more resources per capita showed significant clustering in central Cambodia while areas with fewer resources clustered in the northern and western provinces. Hospital-based inpatient beds, doctors, and nurses were most heavily concentrated in areas of the country with the lowest poverty rates; however, beds and nurses in Non-Hospital Medical Facilities (NHMF) showed increasing concentrations at higher levels of poverty. There is considerable heterogeneity in healthcare resource distribution across Cambodia. Distribution mapping at the local level can inform policy decisions on where to stockpile resources in advance of and for reallocation in the event of a pandemic. These findings will be useful in determining future health resource investment, both for pandemic preparedness and for general health system strengthening, and provide a foundation for future analyses of equity in health services provision for pandemic mitigation planning in Cambodia.


Deterministic direct reprogramming of somatic cells to pluripotency.
Somatic cells can be inefficiently and stochastically reprogrammed into induced pluripotent stem (iPS) cells by exogenous expression of Oct4 (also called Pou5f1), Sox2, Klf4 and Myc (hereafter referred to as OSKM). The nature of the predominant rate-limiting barrier(s) preventing the majority of cells to successfully and synchronously reprogram remains to be defined. Here we show that depleting Mbd3, a core member of the Mbd3/NuRD (nucleosome remodelling and deacetylation) repressor complex, together with OSKM transduction and reprogramming in naive pluripotency promoting conditions, result in deterministic and synchronized iPS cell reprogramming (near 100% efficiency within seven days from mouse and human cells). Our findings uncover a dichotomous molecular function for the reprogramming factors, serving to reactivate endogenous pluripotency networks while simultaneously directly recruiting the Mbd3/NuRD repressor complex that potently restrains the reactivation of OSKM downstream target genes. Subsequently, the latter interactions, which are largely depleted during early pre-implantation development in vivo, lead to a stochastic and protracted reprogramming trajectory towards pluripotency in vitro. The deterministic reprogramming approach devised here offers a novel platform for the dissection of molecular dynamics leading to establishing pluripotency at unprecedented flexibility and resolution.


The medial amygdalar nucleus: a novel glucose-sensing region that modulates the counterregulatory response to hypoglycemia.
To determine whether the medial amygdalar nucleus (MAN) represents a novel brain glucose-sensing region involved in the detection of hypoglycemia and generation of a counterregulatory hormone response. Fura-2 calcium imaging was used to assess glucose responsivity in neurons isolated from the MAN and single-cell real-time reverse transcription PCR used to examine gene expression within glucose-responsive neurons. In vivo studies with local MAN perfusion of the glucoprivic agent, 2-deoxyglucose (2-DG), under normal and hypoglycemic conditions and also after MAN lesioning with ibotenic acid, were used to examine the functional role of MAN glucose sensors. In addition, retrograde neuronal tracer studies were used to examine reciprocal pathways between the MAN and the ventromedial hypothalamus (VMH). The MAN contains a population of glucose-sensing neurons (13.5%), which express glucokinase, and the selective urocortin 3 (UCN3) receptor CRH-R2, but not UCN3 itself. Lesioning the MAN suppressed, whereas 2-DG infusion amplified, the counterregulatory response to hyperinsulinemic hypoglycemia in vivo. However, 2-DG infusion to the MAN or VMH under normoglycemic conditions had no systemic effect. The VMH is innervated by UCN3 neurons that arise mainly from the MAN, and ∼1/3 of MAN UCN3 neurons are active during mild hypoglycemia. The MAN represents a novel limbic glucose-sensing region that contains characteristic glucokinase-expressing glucose-sensing neurons that respond directly to manipulations of glucose availability both in vitro and in vivo. Moreover, UCN3 neurons may provide feedback inhibitory regulation of the counterregulatory response through actions within the VMH and the MAN.


Dynamics of postdecisional processing of confidence.
Most cognitive theories assume that confidence and choice happen simultaneously and are based on the same information. The 3 studies presented in this article instead show that confidence judgments can arise, at least in part, from a postdecisional evidence accumulation process. As a result of this process, increasing the time between making a choice and confidence judgment improves confidence resolution. This finding contradicts the notion that confidence judgments are biased by decision makers seeking confirmatory evidence. Further analysis reveals that the improved resolution is due to a reduction in confidence in incorrect responses, while confidence in correct responses remains relatively constant. These results are modeled with a sequential sampling process that allows evidence accumulation to continue after a choice is made and maps the amount of accumulated evidence onto a confidence rating. The cognitive modeling analysis reveals that the rate of evidence accumulation following a choice does slow relative to the rate preceding choice. The analysis also shows that the asymmetry between confidence in correct and incorrect choices is compatible with state-dependent decay in the accumulated evidence: Evidence consistent with the current state results in a deceleration of accumulated evidence and consequently evidence appears to have a decreasing impact on observed confidence. In contrast, evidence inconsistent with the current state results in an acceleration of accumulated evidence toward the opposite direction and consequently evidence appears to have an increasing impact on confidence. Taken together, this process-level understanding of confidence suggests a simple strategy for improving confidence accuracy: take a bit more time to make confidence judgments.


Microsurgical anatomy of the white matter tracts in hemispherotomy.
Hemispherotomy is a surgical procedure performed for refractory epileptic seizures due to wide hemispheric damage. To describe the microanatomy of the white matter tracts transected in a hemispherotomy and the relationship of the surgical landmarks used during the intraventricular callosotomy. The cortical and subcortical structures were examined in 32 hemispheres. Incision of the temporal stem along the inferior limiting sulcus crosses the insulo-opercular fibers, uncinate, inferior occipitofrontal and middle longitudinal fasciculi, anterior commissure, and optic and auditory radiations. The incision along the superior limiting sulcus transects insulo-opercular fibers and the genu and posterior limb of internal capsule. The incision along the anterior limiting sulcus crosses the insulo-opercular fibers, anterior limb of the internal capsule, anterior commissure, and the anterior thalamic bundle. The disconnection of the posterior part of the corpus callosum may be incomplete if the point at which the last cortical branch of the anterior cerebral artery (ACA) turns upward and disappears from the view through the intraventricular exposure is used as the landmark for estimating the posterior extent of the callosotomy. This ACA branch turns upward before reaching the posterior edge of the splenium in 85% of hemispheres. The falx, followed to the posterior edge of the splenium, is a more reliable landmark for completing the posterior part of an intraventricular callosotomy. The fiber tracts disconnected in hemispherotomy were reviewed. The falx is a more reliable guide than the ACA in completing the posterior part of the intraventricular callosotomy.