Selection of endogenous control and identification of significant microRNA deregulations in cervical cancer.
Cervical cancer causes approximately 350,000 deaths each year. The availability of sensitive and specific diagnostic tests to detect cervical cancer in its early stages is essential to improve survival rates. In this study, we compared two strategies for selecting endogenous controls: miRNA profiling by small-RNA sequencing and a commercially available microfluidic card with 30 recommended endogenous controls preloaded by the manufacturer. We used the RefFinder algorithm and coefficient of variation to select endogenous controls. We selected the combination of miR-181a-5p and miR-423-3p as the most optimal normalizer. In the second part of this study, we determined the differential expression (between tumor/non-tumor groups) of microRNA in cervical cancer FFPE tissue samples. We determined the comprehensive miRNA expression profile using small-RNA sequencing technology and verified the results by real-time PCR. We determined the relative expression of selected miRNAs using the 2-ΔΔCt method. We detected statistically significant upregulation of miR-320a-3p, miR-7704, and downregulation of miR-26a-5p in the tumor group compared to the control group. The combination of these miRNAs may have the potential to be utilized as a diagnostic panel for cervical cancer. Using ROC curve analysis, the proposed panel showed 93.33% specificity and 96.97% sensitivity with AUC = 0.985. We proposed a combination of miR-181a-5p and miR-423-3p as optimal endogenous control and detected potentially significant miRNAs (miR-320a-3p, miR-7704, miR-26a-5p). After further validation of our results, these miRNAs could be used in a diagnostic panel for cervical cancer.


Correlation between Volume of Apical Periodontitis Determined by Cone-beam Computed Tomography Analysis and Endotoxin Levels Found in Primary Root Canal Infection.
This clinical study was conducted to correlate the levels of endotoxins and bacterial counts found in primary endodontic infection with the volume of periapical bone destruction determined by cone-beam computed tomography (CBCT) analysis. Moreover, the levels of bacteria and endotoxins were correlated with the development of clinical features. Twenty-four root canals with primary endodontic disease and apical periodontitis were selected. Clinical features such as pain on palpation, pain on percussion, and previous episode of pain were recorded. The volume (cubic millimeters) of periapical bone destruction was determined by CBCT analysis. Endotoxins and bacterial samplings were collected by using sterile/apyrogenic paper points. Endotoxins were quantified by using limulus amebocyte lysate assay (KQCL test), and bacterial count (colony-forming units [CFU]/mL) was determined by using anaerobic culture techniques. Data were analyzed by Pearson correlation and multiple logistic regression (P < .05). Endotoxins and bacteria were detected in 100% of the root canal samples (24 of 24), with median values of 10.92 endotoxin units (EU)/mL (1.75-128 EU/mL) and 7.5 × 10(5) CFU/mL (3.20 × 10(5)-8.16 × 10(6) CFU/mL), respectively. The median volume of bone destruction determined by CBCT analysis was 100 mm(3) (10-450 mm(3)). The multiple regression analysis revealed a positive correlation between higher levels of endotoxins present in root canal infection and larger volume of bone destruction (P < .05). Moreover, higher levels of endotoxins were also correlated with the presence of previous pain (P < .05). Our findings revealed that the levels of endotoxins found in root canal infection are related to the volume of periapical bone destruction determined by CBCT analysis. Moreover, the levels of endotoxin are related to the presence of previous pain.


Synthesis of Nitrogen Containing Chalcone: A Highly Sensitive and Selective Fluorescent Chemosensor for the Fe3+ Metal Ion in Aqueous Media.
Title compound (PTPO) was synthesized by the reaction of 2-acetyl pyridine with 3,4,5-trimethoxy benzaldehyde. The structure of the compound has been confirmed by spectroscopic techniques. Interaction of chalcone with different metal cations was analyzed based on the fluorescence behavior. Results show that chalcone act as on-off switching fluorescent chemosensor for selective and sensitive detection iron metal ion. Mechanism of quenching and complexation were resolute by Benesi-Hildebrand, Stern-Volmer plot and Job-plot.


Inhibition of noradrenaline release via presynaptic 5-HT1B receptors of the rat vena cava.
In the rat inferior vena cava preincubated with 3H-noradrenaline, the effects of nine serotonin (5-HT) receptor agonists and of eight antagonists (including two beta-adrenoceptor blocking agents) on the electrically evoked 3H overflow were determined. 1. 5-HT, 5-carboxamido-tryptamine, 5-methoxy-3(1,2,3,6-tetrahydropyridine-4-yl)-1H-indole (RU 24969), 5-methoxytryptamine, N,N-dimethyl-5HT, tryptamine and 5-aminotryptamine inhibited the evoked 3H overflow. The potencies of these agonists in inhibiting overflow were significantly correlated with their affinities for 5-HT1B binding sites, but not with their affinities for 5-HT1A, 5-HT1C or 5-HT2 binding sites. 8-Hydroxy-2-(di-n-propylamino)tetralin (8-OH-DPAT), a 5-HT1A receptor agonist, and ipsapirone, a partial agonist at these receptors, did not inhibit overflow. 2. Cyanopindolol facilitated the evoked 3H overflow, an effect which was abolished by propranolol. The maximum inhibition of overflow obtainable with 5-HT was diminished by cyanopindolol. 3. The concentration-response curve for 5-HT was shifted to the right by metitepine, metergoline, quipazine, 6-chloro-2-(1-piperazinyl)pyrazine (MK 212) and propranolol which, given alone, did not affect 3H overflow. The apparent pA2 values of these antagonists tended to be correlated with their affinities for 5-HT1B (but not 5-HT1A, 5-HT1C or 5-HT2) binding sites. Ketanserin, a 5-HT2 receptor antagonist, and spiperone, which blocks 5-HT2 and 5-HT1A but not 5-HT1B or 5-HT1C receptors, failed to antagonize the effect of 5-HT.(ABSTRACT TRUNCATED AT 250 WORDS)


Distribution trends of Salmonella serovars in India (2001-2005).
A total of 3079 samples were received and identified at the National Salmonella and Escherichia Centre (NSEC), Central Research Institute, Kasauli, India during 2001-2005. Out of these, 2098 samples were from humans, 250 from animals, and 726 from meat, vegetables, seafood and the environment. The Salmonella strains isolated were distributed among 35 different Salmonella serovars. The most common serovars from humans were Salmonella Typhi (73%) and Salmonella Paratyphi A (24%) among typhoidal serovars, and Salmonella Worthington (28.2%) and Salmonella Typhimurium (22.5%) among non-typhoidal serovars. The other frequently isolated serovars from different sources were Salmonella Gallinarum, Salmonella Typhimurium, Salmonella Enteritidis and Salmonella Dublin. Analysis of the prevalence of the 10 most common serovars in 18 states indicated that different serovars are distributed in different parts of India. The results from this study may be helpful in formulating preventive strategies to control the spread of infection in the country.


A homogeneous population of lymphokine-activated killer (LAK) cells is incapable of killing virus-, bacteria-, or parasite-infected macrophages.
Previous reports have suggested a role for natural killer (NK) cells in directly lysing host cells infected with bacteria and other intracellular microorganisms. Here, we determined the inability of a highly homogeneous population of lymphokine activated killer (LAK) cells to kill macrophages infected with the following intracellular parasites: Mycobacterium avium, Listeria monocytogenes, Legionella pneumophila, Toxoplasma gondii, and Trypanosoma cruzi. In parallel cytotoxicity assays, LAK cells lysed the tumor targets YAC-1 and P815 effectively. Furthermore, we were able to demonstrate that influenza-specific cytotoxic T lymphocytes (CTL), but not LAK cells, were efficient killers of influenza virus-infected macrophages.


A national analysis of the relationship between hospital factors and post-cardiac arrest mortality.
We sought to generate national estimates for post-cardiac arrest mortality, to assess trends, and to identify hospital factors associated with survival. We used a national sample of US hospitals to identify patients resuscitated after cardiac arrest from 2000 to 2004 to describe the association between hospital factors (teaching status, location, size) and mortality, length of stay, and hospital charges. Analyses were performed using logistic regression. A total of 109,739 patients were identified. In-hospital mortality was 70.6%. A 2% decrease in unadjusted mortality from 71.6% in 2000 to 69.6% in 2004 (OR 0.96, P < 0.001) was observed. Mortality was lower at teaching hospitals (OR 0.58, P = 0.001), urban hospitals (OR 0.63, P = 0.004), and large hospitals (OR 0.55, P < 0.001). Mortality after in-hospital cardiac arrest decreased over 5 years. Mortality was lower at urban, teaching, and large hospitals. There are implications for dissemination of best practices or regionalization of post-cardiac arrest care.


ASTM lights the way for tissue engineered medical products standards: jump start for combination medical products that restore biological function of human tissues.
Everybody hopes for better health and restoration of impaired bodily function, and now that hope is illuminated by the promise of powerful biological tools that make human cells grow and replace human tissue. ASTM Committee F04 on Medical and Surgical Materials and Devices is taking the lead by defining some of those tools as standards that can be used for the development, production, testing, and regulatory approval of medical products.


Imaging of craniopharyngioma.
Craniopharyngiomas are present with a wide range of appearances, but the existence of cysts, calcification, and enhancement in a suprasellar tumor strongly favors the diagnosis. There is a significant differential diagnosis that must be considered. The pre- and postoperative imaging of craniopharyngioma is reviewed.


Epidemiology of acute asthma: IgE antibodies to common inhalant allergens as a risk factor for emergency room visits.
In recent years the morbidity and mortality of asthma has increased, although the etiology is still poorly understood. Most patients with asthma suffer acute attacks that are commonly treated in hospital emergency rooms (ER). In the present study, asthma in adults was studied with acute attacks as a marker for the disease; 102 patients first observed at a university hospital ER with acute airway obstruction were compared to 118 patients observed at the same ER with any diagnosis other than shortness of breath to evaluate allergy as a risk factor for asthma in adults. Sera were assayed for IgE antibody (Ab) to dust mites, cockroach, cat dander, and grass and ragweed pollen. The results demonstrate that in adults younger than 50 years of age, the prevalence of IgE Abs was fourfold greater among subjects with asthma than among control subjects (46/67 versus 12/81; odds ratio, 10.1; 95% confidence interval, 4.9 to 20.7). The population attributable risk for the presence of IgE Ab to one of the five allergens was greater than 50%. Among individuals older than 50 years of age, the prevalence of serum IgE Abs was not significantly increased among patients with acute airway obstruction. In the whole group, the prevalence of IgE Abs to different allergens demonstrated significant seasonal and socioeconomic differences, suggesting that the associated risk is related to exposure to those allergens. The results establish that, with acute attacks of asthma as a marker for adult asthma, the presence of serum IgE Abs to common inhalant allergens is a major risk factor.(ABSTRACT TRUNCATED AT 250 WORDS)


In vivo measurements of relaxivities in the rat kidney cortex.
The aim of this study was to implement a novel noninvasive method to derive the in vivo T1 relaxivity (R1) and T2 relaxivity (R2) in the rat kidney cortex. A two-compartment gadolinium diethylene triamine pentaacetic acid (Gd-DTPA) distribution model was established to estimate the bolus and infusion dosages of Gd-DTPA necessary for obtaining the required steady-state concentration levels. After a single bolus injection of (99m)Tc-DTPA, several blood samples were collected. Based on considerations from the applied two-compartment model, a steady-state concentration was predicted approximately 5-10 minutes after the bolus injection. The plasma concentration levels of Gd-DTPA were measured by simultaneous injection of (99m)Tc-DTPA. Three regions in the cortex (upper, central, and lower) of both rat kidneys were used. A statistical evaluation resulted in the following in vivo relaxivities found at 7 T: R1 = 1.04 +/- 0.08 mM(-1)s(-1) and R2 = 10.78 +/- 0.83 mM(-1)s(-1). Using a 95% confidence interval, no intracortical differences were detected. The relaxivities R1 and R2 calculated in the intact rat kidney cortex were distinctly different from relaxivities found in human plasma: (22 degrees C) 4.42 +/- 0.07 mM(-1)s(-1) (r2> 0.98) and R2 = 5.75 +/- 0.17 mM(-1)s(-1) (r2> 0.98), respectively. The measurements showed a marked difference between in vitro and in vivo relaxivities. Comparison of the distribution rates in pig, human, and rats shows a distinct proportionality between size and renal function.


Venous drainage of the stomach in the domestic rabbit (Oryctolagus cuniculus f. domestica, breed large Chinchilla) and the domestic cat (Felis catus L. f. domestica).
Two basic patterns of venous drainage of the stomach were found in rabbits. In the first, which is highly prevalent (27 cases-90.0%), the v. gastroepiploica dextra is present while no such vein is present in the second one. Other venous channels are represented by the v. gastrica sinistra and by tributaries of the v. lienalis. In cats there are also two basic patterns of the stomach venous drainage. The first highly prevalent type without the v. gastroepiploica dextra was found in 23 cases (76.0%); in the second type this vein was present. In rabbits the v. gastroepiploica sinistra was observed in 25 cases (82.5%), in cats in only two cases (6.6%). The v. gastroepiploica dextra in the rabbit is usually a tributary of the v. portae, while in cats it is a tributary of the v. lienalis. In ten rabbits (33.3%) there was a venous arch around the curvatura ventriculi major, while in cats it was absent. The great difference in the stomach venous pattern in rabbits and cats can be explained by the different zoological position of these two kinds of animals.


Neoadjuvant M-VAC (methotrexate, vinblastine, doxorubicin and cisplatin) effect on the primary bladder lesion.
Of 50 patients with bladder cancer given 1 to 5 cycles of neoadjuvant methotrexate, vinblastine, doxorubicin and cisplatin in a pilot phase I and II study 63 per cent of 41 with pure transitional cell stage T2-4 lesions responded. While significant downstaging occurred by transurethral resection of the bladder in 70 per cent and by cytology in 60 per cent of the patients, the final T response rate by all noninvasive clinical staging procedures, including sonography and computerized tomography, revealed complete remission in 24 per cent and partial remission in 39 per cent. Of 30 patients who underwent pathological staging 33 per cent achieved stage P0 and 17 per cent stage Tis disease or P less than T. Despite extensive re-evaluation by transurethral resection of the bladder and other noninvasive staging procedures, a clinical staging error (T versus P) of 38 per cent was observed. Of the other 9 patients 4 with mixed nontransitional cell histological findings at presentation never achieved complete remission, although 3 had resolution of all transitional cell elements and 5 (10 per cent) were inevaluable. The toxicity of the regimen was generally acceptable but 6 per cent of the patients required hospitalization for neutropenic fever. While this active regimen can clinically (T) and pathologically (P) induce downstaging in a significant number of patients with primary bladder tumors, this pilot study has raised serious questions concerning the design of future nonrandomized and randomized neoadjuvant studies.


Virological surveillance of influenza and other respiratory viruses during six consecutive seasons from 2006 to 2012 in Catalonia, Spain.
Most attention is given to seasonal influenza and respiratory syncytial virus outbreaks, but the cumulative burden caused by other respiratory viruses (RV) is not widely considered. The aim of the present study is to describe the circulation of RV in the general population during six consecutive seasons from 2006 to 2012 in Catalonia, Spain. Cell culture, immunofluorescence and PCR-based assays were used for the RV laboratory-confirmation and influenza subtyping. Phylogenetic and molecular characterizations of viral haemagglutinin, partial neuraminidase and matrix 2 proteins were performed from a representative sampling of influenza viruses. A total of 6315 nasopharyngeal samples were collected, of which 64% were laboratory-confirmed, mainly as influenza A viruses and rhinoviruses. Results show the significant burden of viral aetiological agents in acute respiratory infection, particularly in the youngest cases. The study of influenza strains reveals their continuous evolution through either progressive mutations or by segment reassortments. Moreover, the predominant influenza B lineage was different from that included in the recommended vaccine in half of the studied seasons, supporting the formulation and use of a quadrivalent influenza vaccine. Regarding neuraminidase inhibitors resistance, with the exception of the 2007/08 H275Y seasonal A(H1N1) strains, no other circulating influenza strains carrying known resistance genetic markers were found. Moreover, all circulating A(H1N1)pdm09 and A(H3N2) strains finally became genetically resistant to adamantanes. A wide knowledge of the seasonality patterns of the RV in the general population is well-appreciated, but it is a challenge due to the unpredictable circulation of RV, highlighting the value of local and global RV surveillance.


Immune cell composition in the endometrium of patients with a complete molar pregnancy: Effects on outcome.
In 15% of patients with complete hydatidiform mole (CHM), disease progresses to post-molar gestational trophoblastic neoplasia (GTN) after curettage. Tumor infiltrating lymphocytes (TILs) are essential in overcoming disease in many tumors. Infiltrating lymphocyte composition and density may influence trophoblast regression and development of post-molar GTN. We analyzed immune cell composition and density in curettaged endometrium of patients with CHM which spontaneously regressed, and of patients with CHM which progressed to post-molar GTN. Sixteen patients with CHM and spontaneous regression, and 16 patients with CHM which progressed to post-molar GTN were selected. Immune cell composition and density of natural killer (NK) cells, natural killer T (NKT)-like cells, Cytotoxic T cells, T-Regulatory and T-Helper cells, were determined by multiplex immunohistochemistry (mIHC). Curettaged endometrium of patients with CHM and spontaneous regression contained a slightly higher number of immune cells compared to patients with CHM which progressed to post-molar GTN. NKT-like cell density was significantly higher in patients with spontaneous regression compared to patients with CHM which progressed to post-molar GTN (483 ± 296 vs.295 ± 143 (mean ± SD), p = 0.03) respectively. NKT-like cell density in the spontaneous regression group was split in 'high' and 'low' (i.e. above and below the median number of NKT-like cells). In patients with high NKT-like cell density, hCG normalized earlier than in patients with low NKT-like cell density (9.5 weeks, (range 3.7-14) vs. 12.9 weeks, (range 8.6-17.9), p = 0.05). A high number of NKT-like cells in the endometrium of CHMs may contribute to spontaneous regression of molar trophoblast cells.


Bodies of children in the Institute of Anatomy in Halle (Saale), Germany, between 1920 and 1945.
At the Institute of Anatomy and Cell Biology in Halle (Saale) 74 children's bodies of unknown historical provenance are being held in storage. The aim of this study was the evaluation of their identities, the circumstances of their acquisition, as well as the documentation of their individual characteristics. For these purposes, all bodies were comprehensively examined and photo-documented. Furthermore, CT-scans of 29 bodies were performed and information was collected from various local and national archives. Although most of the bodies were found to be those of stillborn children and infants, five children were between two and twelve years old, according to an age estimate by body-length and carpal bone analysis. The CT-scans revealed the cause of death for some of the children. The embalming method indicates that the bodies date from the first decades of the 20th century, and archival sources containing documents from 1920 to 1960 strongly suggest that these children's bodies were acquired by Institute of Anatomy between 1920 and 1942. During that period, a total of 2602 children's bodies were delivered to the Institute of Anatomy and registered in the communal burial records. At this point, there is no evidence that these children might have been victims of National Socialist crimes. It is planned to give them a dignified burial.


Rapid migration of grafted cortical astrocytes from suspension grafts placed in host thoracic spinal cord.
The cerebral cortices from 14-day gestation rat embryos were prelabeled with Phaseolus vulgaris leucoagglutin (PHAL) and homografted as a cell suspension into host thoracic spinal cord. Animals were sacrificed at 7, 14, 21 and 30 days postimplantation (DPI). Paraffin sections of cervical, thoracic and lumbar spinal cord were double-labeled for the presence of glial fibrillary acidic protein (GFAP) a specific marker for astrocytes, and PHAL, utilized as a marker for graft-derived cells. PHAL-GFAP positive cells were found throughout the thoracic spinal cord at all time periods, indicating that grafted astrocytes migrated along all 3 axes of the host spinal cord (rostral-caudal, dorsal-ventral and left-right). At 30 DPI, graft-derived astrocytes were found in host cervical and lumbar spinal cord. There appeared to be a minimal delay in the onset of migration.


Assessment of postoperative outcomes of hypospadias repair with validated questionnaires.
A standardized assessment for the optimal repair of hypospadias remains elusive. This study utilized validated questionnaires to assess the postoperative functional, cosmetic, and psychosocial outcomes of hypospadias repair. 172 patients who underwent hypospadias repair under the care of a single surgeon were identified. 25 agreed for follow-up using the validated questionnaires of Hypospadias Objective Scoring Evaluation (HOSE), Pediatric Penile Perception Scale (PPPS), and Pediatric Quality of Life Inventory (PedsQL™4.0). Mean follow-up was 59months postoperatively (range 7-113months). Techniques used included tubularized incised plate urethroplasty, meatal advancement and glanuloplasty, and a 2-stage repair. 23 of 25 patients achieved a HOSE score of 14 or more (maximum of 16). The PPPS scores correlated with severity of the hypospadias. Those with glanular hypospadias (mean score=10) scored higher than those with coronal (mean score=9) and penile/penoscrotal hypospadias (mean score=7). There was no correlation between PedsQL™4.0 scores and the severity of hypospadias or procedure used. Validated questionnaires revealed generally good functional, cosmetic, and early psychosocial outcomes after hypospadias repair. The use of validated questionnaires in routine follow-up sessions may facilitate objective assessment of both functional outcomes and patient satisfaction.


Enhancement of transfection by physical concentration of DNA at the cell surface.
Efficient DNA transfection is critical for biological research and new clinical therapies, but the mechanisms responsible for DNA uptake are unknown. Current nonviral transfection methods, empirically designed to maximize DNA complexation and/or membrane fusion, are amenable to enhancement by a variety of chemicals. These chemicals include particulates, lipids, and polymer complexes that optimize DNA complexation/condensation, membrane fusion, endosomal release, or nuclear targeting, which are the presumed barriers to gene delivery. Most chemical enhancements produce a moderate increase in gene delivery and a limited increase in gene expression. As a result, the efficiency of transfection and level of gene expression after nonviral DNA delivery remain low, suggesting the existence of additional unidentified barriers. Here, we tested the hypothesis that DNA transfection efficiency is limited by a simple physical barrier: low DNA concentration at the cell surface. We used dense silica nanoparticles to concentrate DNA-vector (i.e. DNA-transfection reagent) complexes at the surface of cell monolayers; manipulations that increased complex concentration at the cell surface enhanced transfection efficiency by up to 8.5-fold over the best commercially available transfection reagents. We predict that manipulations aimed at optimizing DNA complexation or membrane fusion have a fundamental physical limit; new methods designed to increase transfection efficiency must increase DNA concentration at the target cell surface without adding to the toxicity.


COMFORT™SM communication for oncology nurses: Program overview and preliminary evaluation of a nationwide train-the-trainer course.
The COMFORT Communication Course for Oncology Nurses is a train-the-trainer program funded by the National Cancer Institute (R25) that provides nationwide communication training to improve patient-centered communication in cancer care. The purpose of this article is to provide an overview of the program and present an evaluation of three courses. The curriculum contains seven modules addressing palliative care communication. Pre-course survey of needs, post-course feedback, and follow-up at 6 and 12 months were used to evaluate the program. To date, three courses have been presented to 269 nurses from 34 states and Washington D.C. Post-course evaluations showed high satisfaction with course design, content, and faculty. At 12 months, course participants had implemented institution-wide system changes and communication skill building. On average, each nurse trained 37 other healthcare providers. The COMFORT communication course provides the essential communication skills and tools oncology nurses need to provide quality care across the cancer continuum. Training is needed to prepare oncology nurses with the skills to provide patient-centered communication across the cancer continuum. These skills include training others in communication and implementing process improvement. The COMFORT communication train-the-trainer model is an effective approach to meet this need.


Model biases in rice phenology under warmer climates.
Climate-induced crop yields model projections are constrained by the accuracy of the phenology simulation in crop models. Here, we use phenology observations from 775 trials with 19 rice cultivars in 5 Asian countries to compare the performance of four rice phenology models (growing-degree-day (GDD), exponential, beta and bilinear models) when applied to warmer climates. For a given cultivar, the difference in growing season temperature (GST) varied between 2.2 and 8.2 °C in different trials, which allowed us to calibrate the models for lower GST and validate under higher GST, with three calibration experiments. The results show that in warmer climates the bilinear and beta phenology models resulted in gradually increasing bias for phenology predication and double yield bias per percent increase in phenology simulation bias, while the GDD and exponential models maintained a comparatively constant bias. The phenology biases were primarily attributed to varying phenological patterns to temperature in models, rather than on the size of the calibration dataset. Additionally, results suggest that model simulations based on multiple cultivars provide better predictability than using one cultivar. Therefore, to accurately capture climate change impacts on rice phenology, we recommend simulations based on multiple cultivars using the GDD and exponential phenology models.


Variations in mortality and morbidity by gestational age among infants born at term.
To examine the risks of infant death and neonatal morbidity by week of gestation at term. National U.S. birth cohort study on the basis of singleton live births in 1995-2001 at 37 to 41 completed weeks gestational age (GA), with exclusion of congenital anomalies. Main outcomes included neonatal, postneonatal, and cause-specific infant death; low-Apgar score at 5 minutes; receipt of neonatal mechanical ventilation >or=30 minutes; neonatal seizures; birth injury; and meconium aspiration syndrome. To reduce confounding by indication, we carried out a secondary analysis restricted to low-risk deliveries. In non-Hispanic white women, the mortality rate decreased with increasing GA from 37 to 39 weeks, remained stable from 39 to 40 weeks, and then (for neonatal death) increased at 41 weeks. Rates of low 5-minute Apgar score and mechanical ventilation showed a U-shaped relation across term GAs, and rates of meconium aspiration syndrome and birth injury rose with increasing GA. Results were similar among infants born to low-risk mothers and in non-Hispanic black women. Term infants show considerable heterogeneity across gestational age in neonatal and late infant outcomes, even when born to mothers at low risk. Recent trends toward earlier labor induction may have adverse health impacts.


Aquifer remediation using surfactant-enhanced gas sparging applied to target the contaminant source.
The surfactant-enhanced gas sparging process designed to specifically target the source zone of an organic contaminant in an aquifer with minimal usage of injected additives was investigated using a physical model. Aqueous solutions of the anionic surfactant Sodium dodecylbenzne sulfonate (SDBS) and/or the thickener Sodium carboxymethylcellulose (SCMC) were applied in a contaminated horizontal layer in the simulated laboratory aquifer model followed by gas sparging. Fluorescein sodium salt (FSS) was added to the SDBS/SCMC solutions and represented the organic contaminant. Air and ozone were injected to generate gas sparging. A modified surfactant-enhanced ozone sparging method was also tested by applying additional air venting ports installed in the aquifer above the gas injection zone. Both non-aqueous phase liquid (NAPL) and water-dissolved TCA were applied to the SDBS-applied region to evaluate the removal of contaminants during gas sparging. A significant expansion of the de-saturated zone for the SDBS-applied region was observed during air sparging. During ozone sparging, the fluorescence by FSS in the SDBS-applied layer disappeared over a much wider range than that of the control experiment. SCMC application enhanced the performance of the SDBS-applied gas sparging process. The TCA mass removed by volatilization during air sparging from the SDBS-applied layer was 2.3 times the application in the absence of SDBS. Among five regions of injected NAPL contamination located above the single gas injection port, and during 2 h of ozone sparging, with SDBS applied, more than 50% of fluorescence in the NAPL was removed, whereas under the same conditions with no SDBS applied, less than 30% was removed. Diverted gas flow through the venting ports installed in the aquifer model induced a horizontally expanded oxidative reaction zone during ozone sparging. This study demonstrates enhanced gas sparging performance for the removal of contaminants from the aquifer with limited usage of additives applied specifically to the source zone.


Methohexital-Induced Seizure in a Patient Undergoing Conscious Sedation.
Methohexital is a short-acting barbiturate used for procedural sedation in the emergency department (ED). As with other sedatives, adverse effects with methohexital include excess sedation and hypotension, but this agent can also lower the seizure threshold. We report a patient who developed a generalized seizure after administration of methohexital. A 60-year-old man presented to the ED by ambulance with chest pain and shortness of breath. Paramedics had administered adenosine for supraventricular tachycardia without conversion before arrival to the ED. He had no history of seizures. His initial vital signs in the ED included heart rate of 189 beats/min with a supraventricular rhythm, blood pressure 137/108 mm Hg, respiration 22 breaths/min, and oxygen saturation of 98% on room air. It was decided to attempt synchronized electrical cardioversion, and methohexital 1 mg/kg (120 mg) was administered over 2 min for moderate sedation. Within 15 s of methohexital administration, the patient developed a generalized seizure that lasted for 90 s. After seizure termination, he was successfully cardioverted, returned to his previous baseline level of consciousness within 20 min, and discharged without further problems with a follow-up referral to neurology. WHY SHOULD AN EMERGENCY PHYSICIAN BE AWARE OF THIS?: Methohexital is a short-acting barbiturate used for moderate sedation. Its adverse effects are unique in that it can lower the seizure threshold in some patients. Alternative agents for sedation should be considered in individuals with possible seizure disorders.


Gradual painless visual loss: chronic optic neuropathies.
The clinician must be the ultimate medical detective when dealing with chronic optic neuropathies. History taking is crucial. Clinical examination may require supplementation with visual field testing, fluorescein angiography, ocular and orbital ultrasound imaging, CT and MR imaging, blood test data, and cerebrospinal fluid or tissue biopsy data to determine the specific diagnosis. This supplementation is labor-intensive and time-consuming; the visual loss usually will progress throughout the process, frustrating and frightening the patient and physician. The final common pathway is gradual optic atrophy; the appearance of the optic nerve is rarely adequate to determine the cause of the visual loss. This article includes tables that review diagnostic aids and therapies, and lists the frequency with which several disease entities were encountered over 15 years in one tertiary care neuro-ophthalmic practice. If a specific cause is discernible, then a specific therapy may be available. This approach has the best chance of saving the patient's vision with the least toxicity caused by erroneous trials. By necessity, the work-up for these patients is expensive, but the cost of not pursuing the cause is irrevocable, permanent blindness.


Is clinical experience important for obtaining the primary stability of dental implants with aggressive threads? An ex vivo study.
The aim of this study was to investigate the clinicians' experience on maintaining the primary stability of implants with aggressive threads belonging to a novel dental implant system. Three hundred implants with aggressive threads were inserted in fresh bovine ribs mimicking Type IV bone by five clinicians which were classified according to their previous experience of total number of implant insertion. An independent examiner measured the primary stability of all implants after insertion by using resonance frequency analysis (RFA), electronic percussive testing (EPT) and removal torque methods. No significant differences were detected between the stability values measured by the clinicians (p> 0.05) except the Periotest values (PTVs) of the non-experienced clinician. PTVs of the non-experienced clinician were significantly higher than the PTVs of the expert and good clinicians (p<0.05). Significantly higher stability values were detected in the secondary insertion of the non-experienced clinician as compared to her initial insertion values (p<0.05). No significant differences were detected between the first and second measurements of the other clinicians (p> 0.05). Within the limitations of this ex-vivo study, it may be concluded that experience does not play an important role in maintaining the stability of implants with aggressive threads.


Spatio-temporal patterns of zooplankton community in the Yellow River estuary: Effects of seasonal variability and water-sediment regulation.
Zooplankton community is ecological important because of its high sensitivity to environmental changes especially in estuarine areas. The Yellow River estuary (YRE) in China is the fifth biggest estuary in the world with significant seasonal characteristics and anthropogenic influence of Water-Sediment Regulation (WSR). This study investigated the spatio-temporal patterns of zooplankton in the YRE to explore the response of zooplankton to seasonal variation and WSR. Results suggested that the temporal patterns of zooplankton were mainly characterized by seasonal shift of dominant species. Hierarchical cluster analysis and non-metric multidimensional scaling determined summer, summer-autumn and winter-spring three zooplankton assemblages. Zooplankton spatial distributions represented seasonal consistency, in which the abundance generally showed a decreasing gradient from the river mouth to sea. WSR caused a high species replacement rate in July-August (80.36%) and a dramatic abundance decline from 4224.60 ind./m3 to 1541.10 ind./m3 with persistency and hysteresis effect. The high zooplankton abundance moved seaward in spatial distribution after WSR. Summer spatial pattern was determined with two and three zooplankton station assemblages, which was more clear after WSR. Redundancy analysis identified SSS, SST and transparency as important factors structuring zooplankton spatio-temporal patterns, in which SSS was the key one. The results provide a necessary reference for understanding the response of zooplankton community in estuarine areas to spontaneous changes and anthropogenic factors, and can help the protection of estuarine ecosystems and the formulation of hydrological regulatory policies.


Development of a doxycycline-inducible lentiviral plasmid with an instant regulatory feature.
Lentiviruses provide highly efficient gene delivery vehicles in both dividing and non-dividing cells. Inducible gene expression systems often employ a specific cell line that constitutively expresses a regulatory protein for transgene expression. As one of such inducible expression systems the Tet-On system uses a cell line expressing reverse tetracycline-responsive transcriptional activator (rtTA). The rtTA protein binds to the tetracycline-responsive element (TRE) in the promoter and activates transcription of a transgene in a doxycycline-dependent manner. To establish a universal and instant regulatory system without generating Tet-On cell lines, the cDNAs of rtTA and a testing target gene (PPM1B) were cloned in the bi-directional TRE-containing promoters. Here, we examined whether a basal leaky expression of rtTA allows instantly inducible expression of both rtTA itself and the target gene, PPM1B in a single plasmid using the two mini-CMV promoters. Transient transfection of the lentiviral plasmids into human embryonic kidney HEK293T cells showed a significant induction of PPM1B expression in response to doxycycline, suggesting that these lentiviral plasmids can be used as an instantly inducible mammalian expression vector. However, the expression of rtTA by lentiviral transduction shows a minimal expression without a consistent response to doxycycline, suggesting that the utility of these lentiviral vectors is limited. A potential solution to overcome lentiviral transgene inactivation is proposed.


[Genome-wide association studies].
Within the last year, genome-wide association (GWA) studies have identified a large number of robust associations between genetic variants and common diseases. Two key premises underlie this burst of discovery. First, the HapMap project has provided a catalogue of human genetic variation. Second, genotyping microarrays can now assess up to 1 million SNPs, allowing hypothesis-free screening of the entire genome. An important next step will be to elucidate the mechanisms behind genotype-phenotype associations. Ultimately, the goal is improved prevention, diagnosis and therapy.


Antibiotic resistance patterns and therapeutic outcomes of pediatric Helicobacter pylori infection in a high-migrant Singaporean cohort.
Variation in Helicobacter pylori (H. pylori) disease in terms of prevalence and antibiotic resistance prevails globally requiring a need to develop region-specific surveillance. We aimed to assess the influence of immigration factors upon the interpretation of local Singaporean epidemiological trends in antimicrobial susceptibility patterns and therapeutic outcomes in children with culture-positive H. pylori. We retrospectively analyzed eradication outcomes of children with culture-proven H. pylori infections between 2011 and 2020 at our center, and we also analyzed the antimicrobial susceptibility profiles of the corresponding H. pylori isolates. The cohort was classified into two groups: (1) Native Singaporeans and (2) Non-native Singaporeans (First-/Second-generation immigrants and Non-residents) to correlate with resistance patterns and eradication outcomes. H. pylori culture was done via Kirby-Bauer disk diffusion for the era 2011-2016 and bioMérieux E test for 2016-2020. A total of 70 children (median age 14 [2-17] years) were included in the analysis. 42.9% (30/70) of the cohort displayed some form of antibiotic resistance; clarithromycin resistance was the most prevalent (30.0%), followed by metronidazole (27.5%) and amoxicillin (7.1%). Comparing to natives, non-native Singaporeans were significantly younger at presentation (mean 11.7 vs. 13.7 years, p = 0.043), and a significantly higher proportion of non-natives carried clarithromycin-resistant (51.4% vs. 8.6%, p < 0.001), metronidazole-resistant (47.1% vs. 8.6%, p < 0.001), or multidrug-resistant (resistant to ≥2 drugs) (40.0% vs. 2.9%, p < 0.001] strains. Non-natives were significantly more likely to fail first-line eradication therapy (48.5% failure vs. 23.3%, p = 0.038). The proportion of pan-sensitive H. pylori was significantly lower in first-generation (25.0%, p = 0.001) and second-generation (42.9%, p = 0.018) immigrants compared to natives (82.86%). These conclusions did not vary when the analysis was repeated for each culture method. An antibiotic susceptibility-based approach should be advocated for all patients but especially so for non-natives, who are at higher risk for antimicrobial resistant strains and poorer eradication outcomes.


Outcome for adolescent and young adult patients with osteosarcoma: a report from the Children's Oncology Group.
There are conflicting data regarding age as a prognostic factor in osteosarcoma. The authors conducted a study evaluating the impact of age on prognosis in children and young adults with osteosarcoma enrolled on North American cooperative group trials. Patients with high-grade osteosarcoma of any site enrolled on North American cooperative group trials CCG-7943, POG-9754, INT-0133, and AOST0121 were included in this study. Primary tumor site, age, sex, ethnicity, histologic response, and presence of metastatic disease at diagnosis were evaluated for their impact on overall survival (OS) and event-free survival (EFS). A total of 1054 patients were eligible and had complete data available for the study. Age was not significantly associated with any other presenting covariate analyzed except sex. Age 18 or older was associated with a statistically significant poorer EFS (P = .019) and OS (P = .043). The 10-year EFS and OS in patients <10, 10 to 17, and ≥18 years old were 55%, 55%, 37% and 68%, 60%, 41%, respectively. The poorer EFS in patients ≥18 years old was because of an increased rate of relapse. Presence of metastatic disease at diagnosis, poor histologic response, and pelvic tumor site were also associated with a poorer prognosis. In multivariate analysis, age continued to be associated with poorer EFS (P = .019) and OS (P = .049). In osteosarcoma, age 18 to 30 years is associated with a statistically significant poorer outcome because of an increased rate of relapse. Poorer outcome in adolescent and young adult patients is not explained by tumor location, histologic response, or metastatic disease at presentation.


Rabies and cerebrospinal nematodosis in woodchucks (Marmota monax) from New York.
Abnormal behavior, suggestive of rabies, is not an unusual finding in woodchucks (Marmota monax). Necropsy records, rabies antibody tests, and necropsy examinations performed by the authors indicate that migration of ascarid larvae usually is responsible for the abnormal behavior, and that rabies is an infrequent finding. Baermannization of brain tissue is suggested for a differential diagnosis.


Production and properties of a staphylococcin genetically controlled by the staphylococcal plasmid for exfoliative toxin synthesis.
Previous data from this laboratory showed that certain phage group 2 staphylococci contain a large 56S virulence plasmid containing genes that code for both exfoliative toxin (ET) and a specific staphylococcin. Optimal cultural conditions for bacteriocin production were similar to those found for ET production. The bacteriocin is an extracellular product produced in small quantities that can be neither extracted from cell pellets with 1 M NaCl nor induced with mitomycin C. The staphylococcin is active against a wide variety of gram-positive organisms and also against group 2 staphylococcal strains that have been cured of the plasmid carrying the staphylococcin marker. The bacteriocin is not inactivated by oxidation, mechanical agitation, or boiling for 15 min. It is sensitive to the action of trypsin and Pronase but not lysostaphin and is stable within a pH range of 4 to 9. It has an isoelectric point of approximately 7.7. Removal of the ampholytes and glycerol from electrofocused staphylococcin preparations resulted in total loss of bacteriocin activity.


Ultrastructural observations of delayed implantation in the Japanese long-fingered bat, Miniopterus schreibersii fuliginosus.
Embryonic development in Japanese long-fingered bats proceeded very slowly during and after the delayed implantation period (mid-October to mid-December). The primitive amniotic cavity and endoderm were formed before implantation. At the preimplantation stage (before hibernation) the corpus luteum cells appeared active, but became less active at the implantation stage (in hibernation). Activity was again apparent at the early placentation stage (after arousal).


The contribution of spectral analysis to the diagnosis of carotid artery disease.
Pulsed and continuous-wave Doppler examinations with spectral analysis were performed on 258 carotid arteries in 220 patients and compared with multiplane contrast angiography. Each artery was examined for stenosis and assigned to one of four groups: normal to 19%, 20% to 49%, 50% to 99%, and occlusion. The incidence of disease was 70%. Diagnostic sensitivity with all noninvasive techniques varied from 76% to 90% depending on the degree of stenosis, with an overall accuracy rate of 86%. In an attempt to evaluate the diagnostic contribution of spectral analysis, stenosis classification by peak frequency alone was compared with that of the full laboratory profile, including spectral analysis. In the 147 carotid arteries in which such data were available, diagnostic sensitivity was improved with spectral analysis by 8% to 29% depending on the degree of stenosis. Detection of occlusions was unchanged. The major contribution of spectral analysis was in detecting non-flow-limiting stenoses of 20% to 49%. Spectral analysis added little to the diagnostic ability of peak frequency to detect lesions with greater than 50% stenosis. Spectral analysis of either pulsed Doppler or continuous-wave Doppler signals is an accurate noninvasive method for evaluating carotid bifurcation disease, which particularly improves the detection of non-flow-limiting stenoses.


A Nurse-Led Bridging Program to Reduce 30-Day Readmissions of Older Patients Discharged From Acute Care Units.
Older hospitalized patients are at high risk of early readmissions, requiring the implementation of enhanced coordinated transition programs on discharge. The objective of this study was to evaluate the impact of a nurse-led transition bridging program on the rate of unscheduled readmissions of older patients within 30 days from discharge from geriatric acute care units. A stepped-wedge cluster randomized trial. Seven hundred five patients aged ≥75 years hospitalized in one of 10 acute geriatric units, with at least 2 readmission risk-screening criteria (derived from the Triage Risk Screening Tool), were included from July 2015 to August 2016. The intervention condition consisted in a nurse-led hospital-to-home bridging program with 4 weeks postdischarge follow-up (2 home visits and 2 telephone calls). Unscheduled hospital readmission or emergency department (ED) visits were compared in intervention and control condition within 30 days from discharge. The rate of 30-day readmission or ED visit was 15.5% in the intervention condition vs 17.6% in the control condition [hazard ratio stratified on clusters: 0.61 (upper limit unilateral 95% confidence interval = 1.11), P = .09]. Rate of presence of professional caregivers was increased in the intervention condition (P < .001). Although the intervention resulted in an increase in the rate of implementation of a package of care at the 4-week of follow-up, we could not demonstrate a reduction in the rate of 30-day readmissions or ED visits of older patients at risk of readmission. These findings support the evaluation of this type of program on the longer term.


UvrB protein of Corynebacterium pseudotuberculosis complements the phenotype of knockout Escherichia coli and recognizes DNA damage caused by UV radiation but not 8-oxoguanine in vitro.
In prokaryotic cells, the UvrB protein plays a central role in nucleotide excision repair, which is involved in the recognition of bulky DNA lesions generated by chemical or physical agents. The present investigation aimed to characterize the uvrB gene of Corynebacterium pseudotuberculosis (CpuvrB) and evaluate its involvement in the DNA repair system of this pathogenic organism. In computational analysis, the alignment of the UvrB protein sequences of Escherichia coli, Mycobacterium tuberculosis, Bacillus caldotenax and Corynebacterium pseudotuberculosis showed high similarity and the catalytic amino acid residues and functional domains are preserved. A CpUvrB model was constructed by comparative modeling and presented structural similarity with the UvrB of E. coli. Moreover, in molecular docking analysis CpUvrB showed favorable interaction with EcUvrA and revealed a preserved ATP incorporation site. Heterologous functional complementation assays using E. coli uvrB-deficient cells exposed to UV irradiation showed that the CpUvrB protein contributed to an increased survival rate in relation to those in the absence of CpUvrB. Damaged oligonucleotides containing thymine dimer or 8-oxoguanine lesion were synthesized and incubated with CpUvrB protein, which was able to recognize and excise UV irradiation damage but not 8-oxoguanine. These results suggest that CpUvrB is involved in repairing lesions derived from UV light and encodes a protein orthologous to EcUvrB.


Bacterial assemblages of urban microbiomes mobilized by runoff waters match land use typologies and harbor core species involved in pollutant degradation and opportunistic human infections.
Cities are patchworks of urban catchments divided into functional units according to their commercial, residential and industrial activities, and socio-urbanistic patterns. The hypothesis of city surface microbiomes being structured by socio-urbanistic variables leading to an emergence of synurbic taxa was tested. According to the r/K microbial ecology theory, a gradient of well-adapted synurbic K-strategists and of opportunistic -r-strategists should occur over city surfaces. K-strategists would be core components while r-ones would be transiently detected. To resolve these patterns, sub-catchments (n = 21) of an area of high commercial and industrial activities were investigated over three time periods covering one year. The sub-catchments' land use patterns and associated human behaviors were converted into socio-urbanistic variables and groupings. Bacterial cells mobilized by runoffs per sub-catchment were recovered, and analyzed by classical approaches, microbial source tracking DNA assays and DNA meta-barcoding approaches. Relationships between these datasets, the runoff physico-chemical properties, and descriptors of the socio-urbanistic groupings were investigated. 16S rRNA meta-barcoding analyses showed evidence of the occurrence of K- and r-like strategists. Twenty-eight core genera were identified, and correlation networks revealed large bacterial modules organized around actinobacterial taxa involved in hydrocarbon degradation processes. Other bacterial networks were related to the occurrences of hygienic wastes, and involved bacteria originating from fecal contaminations. Several r-strategists like Sulfurospirillum were recorded and found associated to point source pollutions. The tpm-metabarcoding approach deciphered these r / K strategists at the species level among more than ten genera. Nine core K-like Pseudomomas species were identified. The P. aeruginosa human opportunistic pathogen and P. syringae phytopathogens were part of these K-strategists. Other tpm-harboring bacterial pathogens showed r-like opportunistic distribution patterns. Correlation network analyses indicated a strong incidence of hygienic wastes and hydrocarbon-pollutions on tpm-harboring bacteria. These analyses demonstrated the occurrence of core synurbic bacterial K-strategists over city surfaces.


Description and Validation of Flow-Through Chambers of Respirometry for Measuring Gas Exchange in Animal Trials.
Indirect calorimetry (IC) is a widely used method to study animal energy metabolism by measuring gas exchange. The accuracy of IC depends on detecting variations in signals reflecting the metabolic response, which can be challenging due to measurement noise and external factors. This study proposes a methodology to validate IC systems, including an easy-to-use spreadsheet for data computing, to verify accuracy and detect whole-system leaks. We conducted a recovery test using a simulation of CO2 dynamics in MS Excel and injecting a known CO2 concentration into four respirometry chambers. The thought flow rate of CO2 was observed and compared to the expected rate from the simulation. Data filtering and computing, including a detailed calculation of signals calibration, Bartholomew transformation, and noise reduction, was developed to obtain the gas exchange and heat production parameters using an open-circuit IC system. The results from the recovery test in our system show that the proposed methodology is accurate and precise. The proposed methodology and recovery test can be used to standardize the validation of IC systems together with adequate data computing, providing accurate measurements of animal energy metabolism in different environmental conditions and energy utilization from feeds.


Sheep grazing in the North Atlantic region: A long-term perspective on environmental sustainability.
Sheep grazing is an important part of agriculture in the North Atlantic region, defined here as the Faroe Islands, Greenland, Iceland, Norway and Scotland. This process has played a key role in shaping the landscape and biodiversity of the region, sometimes with major environmental consequences, and has also been instrumental in the development of its rural economy and culture. In this review, we present results of the first interdisciplinary study taking a long-term perspective on sheep management, resource economy and the ecological impacts of sheep grazing, showing that sustainability boundaries are most likely to be exceeded in fragile environments where financial support is linked to the number of sheep produced. The sustainability of sheep grazing can be enhanced by a management regime that promotes grazing densities appropriate to the site and supported by area-based subsidy systems, thus minimizing environmental degradation, encouraging biodiversity and preserving the integrity of ecosystem processes.


Gene trapping uncovers sex-specific mechanisms for upstream stimulatory factors 1 and 2 in angiotensinogen expression.
A single-nucleotide polymorphism (C/A) located within an E-box at the -20 position of the human angiotensinogen (AGT) promoter may regulate transcriptional activation through differential recruitment of the transcription factors upstream stimulatory factor (USF) 1 and 2. To study the contribution of USF1 on AGT gene expression, mice carrying a (-20C) human AGT (hAGT) transgene were bred with mice harboring a USF1 gene trap allele designed to knock down USF1 expression. USF1 mRNA was reduced relative to controls in liver (9 ± 1%), perigenital adipose (16 ± 3%), kidney (17 ± 1%), and brain (34 ± 2%) in double-transgenic mice. This decrease was confirmed by electrophoretic mobility shift assay. Chromatin immunoprecipitation analyses revealed a decrease in USF1, with retention of USF2 binding at the hAGT promoter in the liver of male mice. hAGT expression was reduced in the liver and other tissues of female but not male mice. The decrease in endogenous AGT expression was insufficient to alter systolic blood pressure at baseline but caused reduced systolic blood pressure in female USF1 gene trap mice fed a high-fat diet. Treatment of USF1 knockdown males with intravenous adenoviral short hairpin RNA targeting USF2 resulted in reduced expression of USF1, USF2, and hAGT protein. Our data from chromatin immunoprecipitation assays suggests that this decrease in hAGT is attributed to decreased USF2 binding to the hAGT promoter. In conclusion, both USF1 and USF2 are essential for AGT transcriptional regulation, and distinct sex-specific and tissue-specific mechanisms are involved in the activities of these transcription factors in vivo.


Clinical significance and preventive strategies for contrast-induced nephropathy.
Contrast-induced nephropathy continues to be a common cause of in-hospital acute kidney injury. Published studies on pathogenesis, clinical significance, diagnosis, and preventive measures have dramatically increased significantly in the past several years. This review will focus on new developments in contrast-induced nephropathy. Studies on the clinical significance of contrast-induced nephropathy are reviewed along with initial reports of biomarkers in diagnosing this complication of iodinated contrast administration. Emerging literature on the relative nephrotoxicity of iso-osmolar versus low-osmolar contrast media and the value of bicarbonate hydration are discussed. More recent preventive measures using prostacyclin, 'statins', and erythropoietin are also reviewed. Contrast-induced nephropathy is an increasing cause of acute kidney injury and is associated with significant mortality and morbidity. Future developments in this field will focus on refining the clinical significance of this complication, earlier diagnosis with biomarkers, clarifying the role for bicarbonate and iso-osmolar contrast agents as preventive strategies, and the introduction of new prophylactic techniques on the basis of an improved understanding of pathogenesis at the cellular level.


Characterization of an active spore photoproduct lyase, a DNA repair enzyme in the radical S-adenosylmethionine superfamily.
The major photoproduct in UV-irradiated Bacillus spore DNA is a unique thymine dimer called spore photoproduct (SP, 5-thyminyl-5,6-dihydrothymine). The enzyme spore photoproduct lyase (SP lyase) has been found to catalyze the repair of SP dimers to thymine monomers in a reaction that requires S-adenosylmethionine. We present here the first detailed characterization of catalytically active SP lyase, which has been anaerobically purified from overexpressing Escherichia coli. Anaerobically purified SP lyase is monomeric and is red-brown in color. The purified enzyme contains approximately 3.1 iron and 3.0 acid-labile S(2-) per protein and has a UV-visible spectrum characteristic of iron-sulfur proteins (410 nm (11.9 mM(-1) cm(-1)) and 450 nm (10.5 mM(-1) cm(-1))). The X-band EPR spectrum of the purified enzyme shows a nearly isotropic signal (g = 2.02) characteristic of a [3Fe-4S]1+ cluster; reduction of SP lyase with dithionite results in the appearance of a new EPR signal (g = 2.03, 1.93, and 1.89) with temperature dependence and g values consistent with its assignment to a [4Fe-4S]1+ cluster. The reduced purified enzyme is active in SP repair, with a specific activity of 0.33 micromol/min/mg. Only a catalytic amount of S-adenosylmethionine is required for DNA repair, and no irreversible cleavage of S-adenosylmethionine into methionine and 5'-deoxyadenosine is observed during the reaction. Label transfer from [5'-3H]S-adenosylmethionine to repaired thymine is observed, providing evidence to support a mechanism in which a 5'-deoxyadenosyl radical intermediate directly abstracts a hydrogen from SP C-6 to generate a substrate radical, and subsequent to radical-mediated beta-scission, a product thymine radical abstracts a hydrogen from 5'-deoxyadenosine to regenerate the 5'-deoxyadenosyl radical. Together, our results support a mechanism in which S-adenosylmethionine acts as a catalytic cofactor, not a substrate, in the DNA repair reaction.


Non-Expressed Donor KIR3DL1 Alleles May Represent a Risk Factor for Relapse after T-Replete Haploidentical Hematopoietic Stem Cell Transplantation.
KIR3DL1 alleles are expressed at different levels on the natural killer (NK) cell surface. In particular, the non-expressed KIR3DL1*004 allele appears to be common in Caucasian populations. However, the overall distribution of non-expressed KIR3DL1 alleles and their clinical relevance after T-replete haploidentical hematopoietic stem cell transplantation (hHSCT) with post-transplant cyclophosphamide remain poorly documented in European populations. In a cohort of French blood donors (N = 278), we compared the distribution of expressed and non-expressed KIR3DL1 alleles using next-generation sequencing (NGS) technology combined with multi-color flow cytometry. We confirmed the predominance of the non-expressed KIR3DL1*004 allele. Using allele-specific constructs, the phenotype and function of the uncommon KIR3DL1*019 allotype were characterized using the Jurkat T cell line and NKL transfectants. Although poorly expressed on the NK cell surface, KIR3DL1*019 is retained within NK cells, where it induces missing self-recognition of the Bw4 epitope. Transposing our in vitro observations to a cohort of hHSCT patients (N = 186) led us to observe that non-expressed KIR3DL1 HSC grafts increased the incidence of relapse in patients with myeloid diseases. Non-expressed KIR3DL1 alleles could, therefore, influence the outcome of hHSCT.


Green productivity divergence and factor endowments: Evidence from the Yellow River Basin, China.
Using panel data for the Yellow River Basin (YRB) of China from 2006 to 2017, we investigate cross-city conventional total factor productivity (TFP) and green TFP convergence, and the moderating effects of relative factor endowments on TFP growth. Allowing for cross-city and cross-time variation in the production function, we estimate TFP across cities using a nonlinear varying coefficient model, and decompose it into various input-embedded and input-free productivity components based on the new growth accounting, covering all growth-driving channels. This paper then employs a conditional convergence framework to examine whether convergence occurs, through which channels, and the effects of relative factor endowments on them. Empirical results show that lagging cities fail to achieve TFP catch-up, and that the divergence of capital-embedded and labor-embedded productivity instead triggers a widening of the cross-city TFP gap. Part of the cause of this increase in these gaps is that cities with relatively high capital deepening and capital-to-energy ratio are experiencing rapid TFP growth by driving the quality of capital and labor. Nor have these effects been altered in examining environmentally constrained or green TFP convergence.


Impact of COVID-19 pandemic early response measures on myocardial infarctions and acute cardiac care in Singapore.
The COVID -19 pandemic impacted acute myocardial infarction (AMI) attendances, ST-elevation myocardial infarction (STEMI) treatments, and outcomes. We collated data from majority of primary percutaneous coronary intervention (PPCI)-capable public healthcare centres in Singapore to understand the initial impact COVID-19 had on essential time-critical emergency services. We present data comparisons from 'Before Disease Outbreak Response System Condition (DORSCON) Orange', 'DORSCON Orange to start of circuit breaker (CB)', and during the first month of 'CB'. We collected aggregate numbers of weekly elective PCI from four centres and AMI admissions, PPCI, and in-hospital mortality from five centres. Exact door-to-balloon (DTB) times were recorded for one centre; another two reported proportions of DTB times exceeding targets. Median weekly elective PCI cases significantly decreased from 'Before DORSCON Orange' to 'DORSCON Orange to start of CB' (34 vs 22.5, P = 0.013). Median weekly STEMI admissions and PPCI did not change significantly. In contrast, the median weekly non-STEMI (NSTEMI) admissions decreased significantly from 'Before DORSCON Orange' to 'DORSCON Orange to start of CB' (59 vs 48, P = 0.005) and were sustained during CB (39 cases). Exact DTB times reported by one centre showed no significant change in the median. Out of three centres, two reported significant increases in the proportion that exceeded DTB targets. In-hospital mortality rates remained static. In Singapore, STEMI and PPCI rates remained stable, while NSTEMI rates decreased during DORSCON Orange and CB. The severe acute respiratory syndrome (SARS) experience may have helped prepare us to maintain essential services such as PPCI during periods of acute healthcare resource strain. However, data must be monitored and increased pandemic preparedness measures must be explored to ensure that AMI care is not adversely affected by continued COVID fluctuations and future pandemics.


Right ventricular systolic dysfunction and its prognostic value in heart failure with preserved ejection fraction.
The prognostic impact of right ventricular systolic dysfunction (RVSD) in heart failure (HF) with preserved ejection fraction (HFPEF) is not sufficiently understood. This pilot study evaluates the prevalence and prognostic impact of RVSD in HFPEF. Ninety-five consecutive patients, admitted due to HF within one year were included and followed up for 12 months. Patients were classified based on left ventricular ejection fraction (LVEF) into two groups: HFPEF (LVEF >40%; n = 54), and heart failure with reduced ejection fraction (HFREF) (LVEF < or = 40%; n = 41). RVSD was defined as peak systolic tricuspid annular velocity (S') <10.8 cm/s. The prevalence of RVSD was 22% vs 59%, in HFPEF vs HFREF, respectively (P < 0.001). Patients with HFPEF and RVSD had significantly higher one-year all-cause mortality compared to HFPEF with normal RV function (41.7% vs. 4.8%, P = 0.004). The same trend was found in HFREF (33.3% vs. 5.9%, P = 0.057). A similar outcome was obser ved in cardiovascular mortality (H FPEF 33.3% vs. 0%, P = 0.002 and HFREF 20.8% vs. 0%, P= 0.06). RVSD was the only independent predictor of all-cause one-year mortality in patients with HFPEF (HR 11.5, 95% Cl 2.2 to 59.5, p = 0.004). RVSD is an independent predictor of all-cause mortality in HFPEF. Patients with HFPEF and RVSD had significantly higher one-year all-cause and cardiovascular mortality than those with normal RV function.


Effect of tolbutamide and glyburide on pyruvate kinase flux in isolated rat hepatocytes.
The effects of two representative sulfonylureas, tolbutamide and glyburide, on pyruvate kinase (PK) flux were examined in fasted rat hepatocytes. PK flux was estimated by trapping 14C from NaH14CO3 in a 2 mM lactate pool, accounting for any incomplete trapping by parallel incubations with L-[1-14C]alanine. Glyburide (20 microM) and tolbutamide (1 mM) decreased glucose formation by 34.9% and 54.8%, respectively, from 2 mM lactate. This decrease in glucose formation was associated with a proportional decrease in pyruvate carboxylase (PCOX) flux (32.7% and 50.5%, respectively). Under these conditions, no net change in PK flux was observed. When hepatocytes were preincubated with lactate and/or sulfonylurea addition for 30 min prior to radiolabeling with NaH14CO3, the metabolic state of the cells changed markedly. Glyburide produced a 34.6% decrease in glucose formation and a 31.3% decrease in PCOX flux, but no change in PK flux. In contrast, tolbutamide decreased glucose formation by 12.5% and increased PK flux by 53.2%, but no change in PCOX flux was observed. Such an increase in PK flux may be linked to tolbutamide-mediated increases in fructose-1,6-bisphosphate (F16P) via fructose-2,6-bisphosphate (F26P). These findings demonstrate that tolbutamide and glyburide decrease hepatic glucose production through various alterations in carbohydrate metabolism, depending upon the metabolic state of the cell. In addition, F26P may play a larger role in the hypoglycemic mechanism of action of tolbutamide than glyburide, since pyruvate carboxylase accounted for most of the decrease in glucose formation observed with glyburide and because preincubation with tolbutamide resulted in an activation of PK.


Neuropeptide Y-deficient mice show altered circadian response to simulated natural photoperiod.
Circadian rhythms, endogenously generated by the suprachiasmatic nucleus (SCN), can be synchronized to a variety of photic and non-photic environmental stimuli. Neuropeptide Y (NPY) is produced in the intergeniculate leaflet (IGL) and known to mediate both photic and non-photic influences on the SCN. We recently found that npy-/- mice were slower to shift their locomotor activity onset to the new time of light offset when photoperiod was abruptly changed from light/dark (LD) cycle 18:6 to LD 6:18. In the present study, we measured the locomotor response of npy-/- mice to gradual changes in photoperiod (4 min a day) for 141 days (LD 16:8 changing to LD 8:16), mimicking external LD cycles in nature. When the photoperiod approached LD 8:16, npy-/- mice showed a significantly delayed onset of activity compared to wild-type mice. Activity patterns disintegrated into multiple bouts and intensity of activity decreased as the photoperiod changed and these changes were more pronounced in npy-/- mice. Our results lend further support to the idea that NPY is involved in circadian entrainment responses to seasonal photoperiod changes.


[Possible correlations between the level of alcoholic motivation and the electrophysiological structure of sleep in the rat].
Using modified Porsolt's method, the electrophysiological sleep pattern was studied in normal conditions and after a single intraperitoneal ethanol injection to noninbred male albino rats divided into 2 groups ("high activity" and "low activity" rats). Voluntary alcohol intake in these rats was measured during free choice between 10% ethanol and water for 20 days. "Low activity" rats were characterized by a statistically significant 3.4-fold higher level of ethanol consumption and 2.7-fold longer REM-sleep stage, as compared to "high activity" animals. In "low activity" animals ethanol (1 g/k, 10% solution, i. p.) inhibits and in "high activity" rats it increases REM-sleep stage, thus removing differences in the sleep pattern in the two groups of rats. The data obtained suggest a possible role of REM-sleep in the development of alcohol motivation.


The impacts of plastics on aquatic insects.
Environmental contamination by plastics and its negative effect on biodiversity have been well-documented in several types of organisms, especially in marine environments. Therefore, it is necessary to assess the impacts of plastic on other organisms such as aquatic insects, which predominantly inhabit freshwaters. It is widely known that these organisms are sensitive to environmental change, especially by contamination. Therefore, this study aimed at testing the hypothesis that aquatic insects are impacted by plastic contamination. We made a systematic search for international papers related to plastics and aquatic insects in databases such as Google Scholar, Web of Science, and Scopus. We obtained 1217 studies of which 40 discussed the impacts of contamination by plastics on aquatic insects. We identified two main impacts: the first one is caused by the use of black macroplastic to protect crops from contact with the soil in agriculture. These black macroplastics attract tons of adult aquatic insects (terrestrial stage) that mistake the plastic surface for water because they select oviposition sites through phototaxis or polarotaxis. The second one comes from water contamination that can originate from the inadequate disposal of plastics, which harms young aquatic insects (aquatic phase) when they feed, reproduce, and construct shelters. Our results show the negative impacts of plastics on both larvae and adult aquatic insects. Despite the large knowledge gap regarding the impacts of plastic on aquatic insects, the evidence above is sufficient to consider these organisms important in global discussions regarding the impacts of plastic on biodiversity.


Inherited TNFSF9 deficiency causes broad Epstein-Barr virus infection with EBV+ smooth muscle tumors.
Epstein-Barr virus (EBV) can infect smooth muscle cells causing smooth muscle tumors (SMTs) or leiomyoma. Here, we report a patient with a heterozygous 22q11.2 deletion/DiGeorge syndrome who developed a unique, broad, and lethal susceptibility to EBV characterized by EBV-infected T and B cells and disseminated EBV+SMT. The patient also harbored a homozygous missense mutation (p.V140G) in TNFSF9 coding for CD137L/4-1BBL, the ligand of the T cell co-stimulatory molecule CD137/4-1BB, whose deficiency predisposes to EBV infection. We show that wild-type CD137L was up-regulated on activated monocytes and dendritic cells, EBV-infected B cells, and SMT. The CD137LV140G mutant was weakly expressed on patient cells or when ectopically expressed in HEK and P815 cells. Importantly, patient EBV-infected B cells failed to trigger the expansion of EBV-specific T cells, resulting in decreased T cell effector responses. T cell expansion was recovered when CD137L expression was restored on B cells. Therefore, these results highlight the critical role of the CD137-CD137L pathway in anti-EBV immunity, in particular in the control of EBV+SMT.


Concentrations of unmetabolized folic acid and primary folate forms in pregnant women at delivery and in umbilical cord blood.
The importance of unmetabolized folic acid in maternal and fetal blood is not known. We investigated total folate, tetrahydrofolate (THF), 5-methyltetrahydrofolate (5-MTHF), formyl-THF, 5,10-methenylTHF, and folic acid concentrations in women and in umbilical cord blood at delivery. The study included 87 pregnant women and 29 cord blood samples, including 24 mother-infant pairs. We measured serum concentrations of folate forms by using ultraperformance liquid chromatography-tandem mass spectrometry. Pregnant women who received 400 μg folic acid daily (n = 25) had higher total folate (P = 0.041), 5-MTHF (P = 0.049), and formyl-THF (P < 0.001) concentrations and slightly higher THF (P = 0.093) concentrations than did nonsupplemented pregnant women (n = 61). We measured folic acid concentrations >0.20 nmol/L in 38 (44%) pregnant women and in 55% of the cord serum samples, but these measurements were not explained by maternal supplement use. Concentrations of folic acid were nonsignificantly higher in cord blood from supplemented women than in cord blood from nonsupplemented women (P = 0.154). Proportions of folic acid to total folate in cord serum did not differ according to maternal supplement usage (0.54% compared with 0.43% in supplemented and nonsupplemented women, respectively). Concentrations of folic acid did not differ between maternal and cord serum. However, folic acid constituted a significantly lower proportion of total folate in cord serum than in maternal serum. We detected unmetabolized folic acid in more than one-half of cord blood samples. Folic acid (400 μg/d) supplied during pregnancy is not likely to accumulate in the fetus, in contrast to 5-MTHF and THF, which accumulate in the fetus.


Visualizing Filoviral Nucleoproteins Using Nanobodies Fused to the Ascorbate Peroxidase Derivatives APEX2 and dEAPX.
Fusions of single-domain antibodies (sdAbs, nanobodies) to enzymatic reporters make convenient molecular probes to detect the presence of an antigen of interest. We have previously fused the monomeric hyperactive ascorbate peroxidase derivative APEX2 to anti-Ebolavirus and anti-Marburgvirus sdAbs to generate immunoreagents useful in detecting nucleoprotein (NP) on western blots, ELISA, and within cells following transfection of NP expression plasmids or following virus infection. Here we present the methods used to overexpress and purify these sdAb-APEX2 fusion proteins, and to employ them as probes in various scenarios with colorimetric and fluorometric signal development. We also introduce a dimeric hyperactive ascorbate peroxidase derivative dEAPX that enables bivalent sdAb probes to be produced while avoiding the need to generate more complex tandem sdAbs, leveraging avidity for improved signal strength. The APEX2 and dEAPX reagents appear interchangeable with any existing detection platform and the methods described here should enable a user to study their antigen of interest by simply swapping out the sdAb for their recombinant affinity reagent of choice.


Sialolipoma of Parotid Gland a Rare Histological Presentation.
Sialolipoma is a rare tumour found within both major and minor salivary glands, it is recently described as a variant of salivary gland lipoma. Sialolipoma was first described by Pusiol et al. (Indian J Pathol Microbiol 52(3):379-382, 2009). Sialolipoma is well circumscribed and contains mature adipose tissue admixed with benign salivary gland component. We report a case of 62 years old female who presented with the complaint of swelling in the right parotid region for 5 years duration. Apart from cosmetic reason she had no other complain. All specific investigations done were inconclusive and suggested a benign tumor. Histopathological examination after superficial parotidectomy suggested sialolipoma. As newly described tumor type, care should be taken to distinguish oncocytic sialolipoma from other salivary gland neoplasms such as simple lipoma, pleomorphic adenoma, or oncocytoma.


Brief Report: Coaching Adolescents with Autism Spectrum Disorder in a School-Based Multi-Sport Program.
While physical activity (PA) is often overwhelming for people with ASD, appropriate engagement strategies can result in increased motivation to participate and associated physical and psychosocial benefits. In this framework, the multi-sport Supporting Success program aims to inform good-practice coaching strategies for community coaches to engage with adolescents with ASD in order to foster socialisation. The project employs a community development approach and a Participatory Action Research (PAR) design. Methods include ongoing consultation, focus groups, briefing/debriefing sessions and questionnaire surveys. Preliminary findings indicate that coaching strategies and program design are fundamental variables in the use of sport/PA to help adolescents with ASD to develop social skills and share positive experiences with peers, coaches, educators and local community members.


Perceptual learning can reverse subliminal priming effects.
Masked primes presented prior to a target can result in inverse priming (i.e., benefits on trials in which the prime and the target are mapped onto opposite responses). In five experiments, time-of-task effects on subliminal priming of motor responses were investigated. First, we replicated Klapp and Hinkley's (2002) finding that the priming effect is initially straight (i.e., it benefits congruent trials, in which the prime and targets are mapped onto the same response) or absent, and only later reverses (i.e., faster responses in incongruent than in congruent trials). We show that the presentation of the mask plays a crucial role in this reversal and that the reversal occurs later if the mask pattern is very complex. We suggest that perceptual learning improves the recognition of task-relevant features. Once recognized, these features can trigger the preparation of the alternative response and/or inhibit the prime-activated response. These findings support an active role of the mask in priming.


Hypo- and Hyper-Assembly Diseases of RNA-Protein Complexes.
A key aspect of cellular function is the proper assembly and utilization of ribonucleoproteins (RNPs). Recent studies have shown that hyper- or hypo-assembly of various RNPs can lead to human diseases. Defects in the formation of RNPs lead to 'RNP hypo-assembly diseases', which can be caused by RNA degradation outcompeting RNP assembly. By contrast, excess RNP assembly, either in higher order RNP granules, or due to the expression of repeat-containing RNAs, can lead to 'RNP hyper-assembly diseases'. Here, we discuss the most recent advances in understanding the cause of disease onset, as well as potential therapies from the aspect of modulating RNP assembly in the cell, which presents a novel route to the treatment of these diseases.


A mathematical model and image analysis technique for calculating regional cerebral blood flow.
This paper discusses the theory and implementation of a modular data analysis system for the calculation and imaging of regional cerebral blood flow. The program may be generalized to any system requiring instantaneous and controllable visual data display, and we have set it up for compartmental analysis as described by the Fick equation with certain correction factors. We have included flow charts of the program source code and data paths for the various modules. We also describe the functional hardware components necessary for the execution of these algorithms. Finally, analyzed data are represented from a set of experiments employing differential visual stimulation. The results obtained using our methods are comparable to those in the existing literature; they are more accurately and easily obtained, and have the added power of manipulable digitization of the original image without loss of data.


Gut microbiota-derived nicotinamide mononucleotide alleviates acute pancreatitis by activating pancreatic SIRT3 signalling.
Gut microbiota dysbiosis induced by acute pancreatitis (AP) exacerbates pancreatic injury and systemic inflammatory responses. The alleviation of gut microbiota dysbiosis through faecal microbiota transplantation (FMT) is considered a potential strategy to reduce tissue damage and inflammation in many clinical disorders. Here, we aim to investigate the effect of gut microbiota and microbiota-derived metabolites on AP and further clarify the mechanisms associated with pancreatic damage and inflammation. AP rat and mouse models were established by administration of caerulein or sodium taurocholate in vivo. Pancreatic acinar cells were exposed to caerulein and lipopolysaccharide in vitro to simulate AP. Normobiotic FMT alleviated AP-induced gut microbiota dysbiosis and ameliorated the severity of AP, including mitochondrial dysfunction, oxidative damage and inflammation. Normobiotic FMT induced higher levels of NAD+ (nicotinamide adenine dinucleotide)-associated metabolites, particularly nicotinamide mononucleotide (NMN). NMN administration mitigated AP-mediated mitochondrial dysfunction, oxidative damage and inflammation by increasing pancreatic NAD+ levels. Similarly, overexpression of the NAD+ -dependent mitochondrial deacetylase sirtuin 3 (SIRT3) alleviated the severity of AP. Furthermore, SIRT3 deacetylated peroxiredoxin 5 (PRDX5) and enhanced PRDX5 protein expression, thereby promoting its antioxidant and anti-inflammatory activities in AP. Importantly, normobiotic FMT-mediated NMN metabolism induced SIRT3-PRDX5 pathway activation during AP. Gut microbiota-derived NMN alleviates the severity of AP by activating the SIRT3-PRDX5 pathway. Normobiotic FMT could be served as a potential strategy for AP treatment.


Fungal and aflatoxin contamination of medicinal herbs.
Since the consumption of aromatic and medicinal herbs has been increasing in the last years, the Argentinian Health Authorities are concerned to control the quality and security of them. Fungal and aflatoxin contamination are two parameters to be taken into account, to ensure the harmlessness of the phytomedicinal products. In 81 different samples, grouped in end products (EP), raw material (RM) and at harvest (SH), fungal flora (enumeration and identification) as well as naturalAspergillus flavus and aflatoxin occurrence were investigated. In all samples fungal counts fulfilled the international general recommendation limits (maximum 10(5) cfu/g). Predominant flora was made up by xerophilic species ofAspergillus(100%), byPeniciIlium (< 50%) and in less percentage byFusarium (5.6%). Among the Aspergilli, A.flavus was present in all the three groups of samples. Using a TLC method, 47% of A. flavus isolates were toxinogenic, producing aflatoxin B1 and B2. In herbs, 4.7% of RM samples were naturally contaminated with aflatoxins B1 and B2. Considering the carcinogenic activity of aflatoxins it is essential to regulate them in the raw material (vegetal drug).


Evaluation of inflammatory reactions and genotoxic effects after exposure of nasal respiratory epithelia to benzene.
The aim of this study was to identify inflammatory changes as well as genotoxic effects in cultivated human respiratory epithelial cells after in vitro exposure to benzene. Primary cell cultures of nasal respiratory mucosa were exposed to synthetic air enriched with 5,000 microg/m(3) of benzene at an air/liquid interface over 8 h and then to synthetic air only over the following 24 h. Controls were continuously exposed to synthetic air over 32 h. To detect inflammatory reactions, release of prostaglandin E(2) was quantified using a competitive enzyme immunoassay. The Comet Assay was used to quantify the ratio of apoptotic cells with benzene-induced DNA fragmentation. Prostaglandin release as well as DNA fragmentation increased after 8 h of exposure and remained elevated throughout the following 24 h but did not increase in controls. High concentrations of benzene induce an inflammatory response and possibly fragmentation of DNA in respiratory epithelial cells. These findings have to be discussed with respect to possible mutagenic or carcinogenic effects of benzene in nasal respiratory cells.


Snare cautery debridement prior to Nd:YAG photoablation improves treatment efficiency of broad-based adenomas of the colorectum.
Forty-six patients who underwent piecemeal resection of large, sessile villous adenomas followed by immediate laser photoablative therapy are presented. In contrast to previously published series utilizing laser alone, prior snare resection of such lesions provides copious tissue for histologic analysis, decreases the amount of residual neoplasm requiring laser treatment, and has the potential to improve treatment efficacy.


A new paradigm for human resuscitation research using intelligent devices.
To develop new methods for studying correlations between the performance and outcome of resuscitation efforts in real-world clinical settings using data recorded by automatic devices, such as automatic external defibrillators (AEDs), and to explore effects of shock timing and chest compression depth in the field. In 695 records of AED use in the pre-hospital setting, continuous compression data were recorded using AEDs capable of measuring sternal motion during compressions, together with timing of delivered shocks and the electrocardiogram. In patients who received at least one shock, putative return of spontaneous circulation (P-ROSC) was defined as a regular, narrow complex electrical rhythm > 40 beats/min with no evidence of chest compressions at the end of the recorded data stream. Transient return of spontaneous circulation (t-ROSC) was defined as the presence of a post-shock organized rhythm > 40 beats/min within 60s, and sustained > or = 30 s. 2x2 contingency tables were constructed to examine the association between these outcomes and dichotomized time of shock delivery or chest compression depth, using the Mood median test for statistical significance. The probability of P-ROSC for first shocks delivered < 50 s (the median time) after the start of resuscitation was 23%, versus 11% for first shocks > 50 s (p=0.028, one tailed). Similarly, the probability of t-ROSC for shorter times to shock was 29%, compared to the 15% for delayed first shocks (p=0.016). For shocks occurring > 3 min after initiation of rescue attempts, the probability of t-ROSC with pre-shock average compression depth > 5 cm was more than double that with compression depth < 5 cm (17.7% vs. 8.3%, p=0.028). For shocks > 5 min, the effect of deeper compressions increased (23.4% versus 8.2%, p=0.008). Much can be learned from analysis of performance data automatically recorded by modern resuscitation devices. Use of the Mood median test of association proved to be sensitive, valid, distribution independent, noise-resistant and also resistant to biases introduced by the inclusion of hopeless cases. Efforts to shorten the time to delivery of the first shock and to encourage deeper chest compressions after the first shock are likely to improve resuscitation success. Such refinements can be effective even after an unknown period of preceding downtime.


Molecular mechanism of acceptor selection in cyclodextrin glycosyltransferases catalyzed ginsenoside transglycosylation.
Understanding the mechanisms of enzyme specificity is increasingly important from a fundamental viewpoint and for practical applications. Transglycosylation has attracted many attentions due to its importance in improving the functional properties of acceptor substrates both in vivo and in vitro. Cyclodextrin glucanotransferase (CGTase) is one of the key enzymes in transglycosylation, it has a broad substrate spectrum and utilizes sugar as the donor. However, little is known about the acceptor selectivity of CGTase, which greatly hampers efforts toward the rational design of desirable transglycosylated derivatives. In this study, we found that the CGTase from Bacillus circulans, BcCGTase, was able to form glycosylated products with diverse ginsenosides. In particular, it not only carries out diverse mono-, di-, and even higher-order glycosylations via the transfer of glucose moieties to the COGlc positions, but also can glycosylate the C3-OH position of ginsenosides. In contrast, another CGTase from Bacillus licheniformis (BlCGTase) showed relatively specific acceptor preference with only several ginsenosides. Structural comparison between BcCGTase and BlCGTase revealed that the Arg74/K81 position within the acceptor-binding sites of BcCGTase/BlCGTase was responsible for the differences in catalytic specificity for ginsenoside F1. Further mutagenesis confirmed their roles in the acceptor selection. In conclusion, our study not only demonstrates the acceptor selectivity of CGTases, but also provides insight into the catalytic mechanism of CGTases, which will potentially increase the utility of CGTase for biosynthesis of new, rationally designed transglycosylated derivatives.


Impact assessment of the European Clinical Trials Directive: a longitudinal, prospective, observational study analyzing patterns and trends in clinical drug trial applications submitted since 2001 to regulatory agencies in six EU countries.
Shifts in clinical trial application rates over time indicate if the attractiveness of a country or region for the conduct of clinical trials is growing or decreasing. The purpose of this observational study was to track changes in drug trial application patterns across several EU countries in order to analyze the medium-term impact of the EU Clinical Trials Directive 2001/20/EC on the conduct of drug trials. Rates of Clinical Trial Applications (CTA) for studies with medicinal products in those six countries in the EU, which authorize on average more than 500 trials per year, were analyzed. Publicly available figures on the number of annually submitted CTA, the distribution of trials per phase and the type of sponsorship were tracked; missing data were provided by national drug agencies. Since 2001, the number of CTA in Italy and Spain increased significantly (5.0 and 2.5% average annual growth). For Italy, the gain was driven by a strong increase of applications from academic trial sponsors; Spain's growth was due to a rise in trials run by commercial sponsors. The Netherlands, Germany, France and the UK saw a decline (1.9, 2.3, 3.0 and 5.3% average annual diminution; significant (P < 0.05) except for Germany) in clinical drug trials. The decrease in the UK was caused by a sharp fall in academic trial activities. Across the six analyzed countries, no EU-wide trial-phase-specific patterns or trends were observed. The EU Clinical Trials Directive 2001/20/EC did not achieve the harmonization of clinical trial requirements across Europe. Rather, it resulted in the leveling of clinical trial activities caused by a continuing decrease in CTA rates in the Netherlands, Germany, France and the UK. Southern European countries, Italy and Spain, benefited to some extent from policy changes introduced by the Directive. In Italy's case, national funding measures helped to considerably promote the conduct of non-commercial trials. On the other hand, the EU Directive-driven transition from liberal policy environments, based on non-explicit trial approval through notifications, towards red-taped processes of trial authorization, contributed to the decreases in trial numbers in Germany and the UK. In the latter case, national research governance concerns had a share in the country's marked decline. However, different EU member states successfully developed best practices, which a new European legislation should take into consideration to resume Europe's attractiveness and international competitiveness for the conduct of clinical trials.


Altered functional connectivity of the amygdala and its subregions in typhoon-related post-traumatic stress disorder.
New evidence suggests that the centromedial amygdala (CMA) and the basolateral amygdala (BLA) play different roles in threat processing. Our study aimed to investigate the effects of trauma and post-traumatic stress disorder (PTSD) on the functional connectivity (FC) of the amygdala and its subregions. Twenty-seven patients with typhoon-related PTSD, 33 trauma-exposed controls (TEC), and 30 healthy controls (HC) were scanned with a 3-Tesla magnetic resonance imaging scanner. The FCs of the BLA, the CMA, and the amygdala as a whole were examined using a seed-based approach, and then, the analysis of variance was used to compare the groups. We demonstrated that the BLA had a stronger connectivity with the prefrontal cortices (PFCs) and angular gyrus in the PTSD group than in the TEC group. Additionally, compared with the PTSD and the HC groups, the TEC group exhibited decreased and increased BLA FC with the ventromedial PFC and postcentral gyrus (PoCG), respectively. Furthermore, the PTSD group showed abnormal FC between the salience network and default-mode network, as well as the executive control network. Compared with the HC group, the TEC group and the PTSD group both showed decreased BLA FC with the superior temporal gyrus (STG). Finally, the FCs between the bilateral amygdala (as a whole) and the vmPFC, and between the BLA and the vmPFC have a negative correlation with the severity of PTSD. Decreased BLA-vmPFC FC and increased BLA-PoCG FC may reflect PTSD resilience factors. Trauma leads to decreased connectivity between the BLA and the STG, which could be further aggravated by PTSD.


Phytochemical composition and antioxidant activity of Cinnamomum burmannii Blume extracts and their potential application in white chocolate.
This study aims at determining the potentials of cinnamon (Cinnamomun burmannii) extracts to improve the health-promoting properties of white chocolate. LC-HRMS analysis was employed to obtain information regarding the phytochemical content while the phosphomolybdenum, FRAP and DPPH assays were used to determine antioxidant activity of cinnamon extract. Furthermore, the cinnamon extract was loaded into nanoparticles before adding it to white chocolate. The results show that cinnamon extracts contained phenols up to 310 mg EE and possessed antioxidant activity up to 260 mg TAE per gram of dry extract depending on the extraction mode (i.e., traditional and ultrasonic-assisted method) and the solvent type. The cinnamon extract contained catechin, epicatechin, procyanidin B2, quercitrin, 3,4-dihydroxybenzaldehyde, protocatechuic acid and cinnamic acid at levels of 51, 53, 1396, 13, 1138, 228 and 934 µg/g of dry extract, respectively. The encapsulated cinnamon extract increased the phenolic content of white chocolate from 47.6 to 1060.6 µg EE/g.


[A case of both lung giant bullae treated by thoracoscopic bilateral resection].
We experienced a case of multiple bilateral giant bullae of the lungs and treated by thoracoscopic bilateral resection. A 46-year-old male was admitted to our hospital on the diagnosis of bilateral giant bullae of the lungs. Chest CT scan and lung perfusion scintigraphy showed giant bullae at the apex of both lungs and at the left lower lobe. The border of the bullae was relatively clear, and the other lung was almost normal although it was compressed by the bullae. We evaluated this case and found that it was a good indication for a thoracoscopic bilateral resection. The patient was positioned on his back, and a successful thoracoscopic bilateral resection was performed. The patient was discharged from the hospital on the 10th day postoperatively. Chest CT scan, lung perfusion scintigraphy and respiratory function test performed 2 months postoperatively, showed remarkable improvement. We would recommend this surgical technique in patients with similar diagnosis.