A randomised trial of three or six courses of etoposide cyclophosphamide methotrexate and vincristine or six courses of etoposide and ifosfamide in small cell lung cancer (SCLC). I: Survival and prognostic factors. Medical Research Council Lung Cancer Working Party.
A total of 458 eligible patients, from 21 centres, with histologically or cytologically confirmed SCLC were allocated at random to three chemotherapy regimens, each given at 3-week intervals. In two regimens, etoposide, cyclophosphamide, methotrexate and vincristine were given for a total of either three courses (ECMV3) or six courses (ECMV6). In the third regimen, etoposide and ifosfamide were given for six courses (EI6). Patients with limited disease (56% of the total) also received radiotherapy to the primary site after the third course of chemotherapy in all three groups. A partial response occurred in 45% of 144 ECMV3 patients, 48% of 141 ECMV6, and 53% of 141 EI6 patients assessed, and a complete response in a further 15%, 9%, and 13% respectively, giving total response rates of 60%, 57%, and 67%, respectively. There was no overall survival advantage to any of the three regimens. At 1 year, 24%, 29%, and 30% of patients were alive, and at 2 years 7%, 8%, and 9%, respectively. The median survival time was 7.4 months in the ECMV3 group, 8.6 months in the ECMV6 group and 8.8 months in the EI6 group. The individual factors: poor performance status, extensive disease, the presence of dysphagia and a raised white blood cell count on admission adversely affected prognosis. The results do not exclude the possibility of a minor survival advantage with the two 6-course regimens. The findings on quality of life are presented in the companion paper (MRC Lung Cancer Working Party, 1993b).


CARD9-mediated ambient PM2.5-induced pulmonary injury is associated with Th17 cell.
Inflammation and oxidative stress are important risk factors in PM2.5-induced injury. The current study attempted to study the role of caspase recruitment domain (CARD) 9 in ambient PM2.5-induced pulmonary injury in mice. The CARD9-/- and C57BL/6 mice were exposed to 3.6, 7.8 and 15.6mg/kgbw of PM2.5 or saline by intratracheal instillation. After the last PM2.5 exposure, the spleen, bronchoalveolar lavage fluid (BALF) and pulmonary tissue were collected and examined. The results showed that PM2.5 exposure induced inflammatory cell infiltration and alveolar septal thickening in the lung. Regulatory T (Treg) cells in spleen of CARD9-/- mice were significantly higher than that in the C57BL/6 mice, whereas the T helper cells 17 (Th17) were lower. The levels of interleukin (IL)-6 and IL-17A in BALF of the CARD9-/- mice were significantly lower than that in the C57BL/6 mice. The mRNA expression of IL-17A, IL-6 and RORγt in the lung of the CARD9-/- mice is significantly lower than that in the C57BL/6 mice, whereas the mRNA levels of Foxp3 in CARD9-/- mice were significantly higher than that in the C57BL/6 mice at the same dose of PM2.5. The protein expression of nuclear factor κB (NF-κB) is higher in the C57BL/6 mice than that in the CARD9-/- mice. This study indicates that ambient PM2.5-induced pulmonary injury is associated with the immune and inflammatory response. CARD9-mediated Th17/Treg differentiation probably played an important role in PM2.5-induced pulmonary damage.


Neurofibrillary degeneration in a case of quadriplegia and myoclonic movement.
A case of spastic quadriplegia with myoclonic movement was studied by both light and electron microscope. Argentophilic inclusions were found in a unique distribution involving the cerebral cortex, brain stem, cerebellum, and the spinal cord, including the motor neuron system. The fine structure of the inclusions consisted of accumulation of previously undescribed straight or wavy tubules approximately 120A wide devoid of periodic constriction.


Poly(A) tail synthesis and regulation: recent structural insights.
Polyadenylation at the 3' ends of mRNAs is critical to the translation and stability of the messages. Recently determined structures of poly(A) polymerase, U1A and domains of the poly(A)-binding protein provide a framework for understanding the synthesis and regulation of the poly(A) tail.


Optimizing appointment template and number of staff of an OB/GYN clinic--micro and macro simulation analyses.
The Department of Obstetrics and Gynecology (OB/GYN) at the University of Arkansas for Medical Sciences (UAMS) tested various, new system-restructuring ideas such as varying number of different types of nurses to reduce patient wait times for its outpatient clinic, often with little or no effect on waiting time. Witnessing little progress despite these time-intensive interventions, we sought an alternative way to intervene the clinic without affecting the normal clinic operations. The aim is to identify the optimal (1) time duration between appointments and (2) number of nurses to reduce wait time of patients in the clinic. We developed a discrete-event computer simulation model for the OB/GYN clinic. By using the patient tracker (PT) data, appropriate probability distributions of service times of staff were fitted to model different variability in staff service times. These distributions were used to fine-tune the simulation model. We then validated the model by comparing the simulated wait times with the actual wait times calculated from the PT data. The validated model was then used to carry out "what-if" analyses. The best scenario yielded 16 min between morning appointments, 19 min between afternoon appointments, and addition of one medical assistant. Besides removing all peak wait times and bottlenecks around noon and late in the afternoon, the best scenario yielded 39.84 % (p<.001), 30.31 % (p<.001), and 15.12 % (p<.001) improvement in patients' average wait times for providers in the exam rooms, average total wait time at various locations and average total spent time in the clinic, respectively. This is achieved without any compromise in the utilization of the staff and in serving all patients by 5 pm. A discrete-event simulation model is developed, validated, and used to carry out "what-if" scenarios to identify the optimal time between appointments and number of nurses. Using the model, we achieved a significant improvement in wait time of patients in the clinic, which the clinic management initially had difficulty achieving through manual interventions. The model provides a tool for the clinic management to test new ideas to improve the performance of other UAMS OB/GYN clinics.


Yoda1 and phosphatidylserine exposure in red cells from patients with sickle cell anaemia.
Phosphatidylserine (PS) exposure is increased in red cells from sickle cell anaemia (SCA) patients. Externalised PS is prothrombotic and attractive to phagocytes and activated endothelial cells and thus contributes to the anaemic and ischaemic complications of SCA. The mechanism of PS exposure remains uncertain but it can follow increased intracellular Ca2+ concentration ([Ca2+]i). Normally, [Ca2+]i is maintained at very low levels but in sickle cells, Ca2+ permeability is increased, especially following deoxygenation and sickling, mediated by a pathway sometimes called Psickle. The molecular identity of Psickle is also unclear but recent work has implicated the mechanosensitive channel, PIEZO1. We used Yoda1, an PIEZO1 agonist, to investigate its role in sickle cells. Yoda1 caused an increase in [Ca2+]i and PS exposure, which was inhibited by its antagonist Dooku1 and the PIEZO1 inhibitor GsMTx4, consistent with functional PIEZO1. However, PS exposure did not necessitate an increase in [Ca2+]i. Two PKC inhibitors were also tested, chelerytherine chloride and calphostin C. Both reduced PS exposure whilst chelerytherine chloride also reduced Yoda1-induced increases in [Ca2+]i. Findings are therefore consistent with the presence of PIEZO1 in sickle cells, able to mediate Ca2+ entry but that PKC was also involved in both Ca2+ entry and PS exposure.


Enhanced performance of PVDF nanocomposite membrane by nanofiber coating: A membrane for sustainable desalination through MD.
Membrane distillation (MD) is a promising separation technique capable of being used in the desalination of marine and brackish water. Poly(vinylidene fluoride) (PVDF) flat sheet nano-composite membranes were surface modified by coating with electro-spun PVDF nano-fibres to increase the surface hydrophobicity. For this purpose, the nano-composite membrane containing 7 wt.% superhydrophobic SiO2 nano-particles, which showed the highest flux in our previous work, was first subjected to pore size augmentation by increasing the concentration of the pore forming agent (Di-ionized water). Then, the prepared flat sheet membranes were subjected to nanofibres coating by electro-spinning. The uncoated and coated composite fabricated membranes were characterized using contact angle, liquid entry pressure of water, and scanning electron microscopy. The membranes were further tested for 6 h desalination by direct contact membrane distillation (DCMD) and vacuum membrane distillation (VMD), with a 3.5 wt.% synthetic NaClaq as the feed. In DCMD the feed liquid and permeate side temperature were maintained at 27.5 °C and 15 °C, respectively. For VMD, the feed liquid temperature was 27 °C and a vacuum of 94.8 kPa was applied on the permeate side. The maximum permeate flux achieved was 3.2 kg/m(2).h for VMD and 6.5 kg/m(2).h for DCMD. The salt rejection obtained was higher than 99.98%. The coated membranes showed a more stable flux than the uncoated membranes indicating that the double layered membranes have great potential in solving the pore wetting problem in MD.


To see and remember: visually specific information is retained in memory from previously attended objects in natural scenes.
What is the nature of the representation formed during the viewing of natural scenes? We tested two competing hypotheses regarding the accumulation of visual information during scene viewing. The first holds that coherent visual representations disintegrate as soon as attention is withdrawn from an object and thus that the visual representation of a scene is exceedingly impoverished. The second holds that visual representations do not necessarily decay upon the withdrawal of attention, but instead can be accumulated in memory from previously attended regions. Target objects in line drawings of natural scenes were changed during a saccadic eye movement away from those objects. Three findings support the second hypothesis. First, changes to the visual form of target objects (token substitution) were successfully detected, as indicated by both explicit and implicit measures, even though the target object was not attended when the change occurred. Second, these detections were often delayed until well after the change. Third, changes to semantically inconsistent target objects were detected better than changes to semantically consistent objects.


Sequestration of anti-Cra in the placenta: serologic demonstration by placental elution.
Few cases of antibodies to the Cromer (a) antigen have been described during pregnancy. Interestingly, the anti-Cr(a) titers decreased during pregnancy, and although the newborns were Cr(a)+, the direct antiglobulin tests (DATs) were negative and hemolytic disease of the newborn (HDN) was not observed. Cromer antigens reside on the decay-accelerating factor (DAF), which is expressed on the fetal derived syncytiotrophoblast layer of the placenta. It has been postulated that Cromer antibodies are not transported to the fetus, but are bound to placental DAF, thereby protecting the fetus from HDN and causing the disappearance of Cromer antibody in maternal plasma. This report is the first to demonstrate Cromer antibody sequestration by the placenta. A G4P1 woman with an anti-Cr(a) presented for prenatal care during her fourth pregnancy. The anti-Cr(a) titer decreased from 64 at 7 weeks gestation to undetectable after 25 weeks. At delivery, the infant had no evidence of HDN. The infant's DAT was negative, and the maternal plasma, cord plasma, and the cord blood eluate were negative with screening cells, the infant's cord cells, and the mother's cells. Placental eluates revealed anti-Cr(a). This is the fourth case report of a Cromer (a-) woman producing anti-Cr(a) during pregnancy, and the first demonstrating anti-Cr(a) sequestration in the placenta. The presence of anti-Cr(a) in the placental eluate, but not in the cord plasma, maternal plasma, or cord blood eluate, strongly supports the hypothesis that DAF at the fetomaternal interface absorbs anti-Cr(a) from the maternal circulation blocking its passage to the fetus.


Short- and long-term impacts of the COVID-19 epidemic on urban PM2.5 variations: Evidence from a megacity, Chengdu.
As the new coronavirus pandemic enters its third year, its long-term impact on the urban environment cannot be ignored, especially in megacities with more than millions of people. Here, we analyzed the changes in the concentration levels, emission sources, temporal variations and holiday effects of ambient fine particulate matter (PM2.5) and its chemical components in the pre- and post-epidemic eras based on high-resolution, long time-series datasets of PM2.5 and its chemical components in Chengdu. In the post-epidemic era, the PM2.5 concentration in Chengdu decreased by 7.4%, with the components of PM2.5 decreasing to varying degrees. The positive matrix factorization (PMF) results indicated that the emissions from soil dust and industrial production were significantly lower during the COVID-19 lockdown period and post-epidemic era than those in the pre-epidemic era. In contrast, the contribution of secondary aerosols to PM2.5 during these two periods increased by 2.7% and 6.6%, respectively. Notably, we found that PM2.5 and its components substantially decreased on workdays and holidays in the post-epidemic era due to the reduced traffic volume and outdoor activities. This provides direct evidence that changes in the habitual behavior patterns of urban residents in the post-epidemic era could exert an evident positive impact on the urban environment. However, the higher PM2.5 concentration was observed due to the increased consumption of regular (As4S4, Xionghuang in Chinese) and "sulfur incense" during the Dragon Boat Festival holiday in the post-epidemic era. Finally, we examined the potential effects of sporadic COVID-19 outbreaks on the PM2.5 concentration in Chengdu, and there was no decrease in PM2.5 during two local COVID-19 outbreak events due to the strong influence of secondary pollution processes.


Electrokinetic remediation of fluorine-contaminated soil: conditioning of anolyte.
The feasibility of anolyte conditioning on electrokinetic remediation of fluorine-contaminated soil was investigated with a field soil. The initial concentration of fluorine, pH and water content in the soil were 414mg/kg, 8.91 and 15%, respectively. Because the extraction of fluorine generally increased with the soil pH, the pH of the anode compartment was controlled by circulating strong alkaline solution to enhance the extraction of fluorine during electrokinetic remediation. The removal of fluorine increased with the concentration of the alkaline solution and applied current density and fluorine removed up to 75.6% within 14 days. Additionally, anolyte conditioning sharply increased the electro-osmotic flow, which enhanced the removal of fluorine in this study. In many respects, anolyte conditioning in electrokinetic remediation of fluorine-contaminated soil will be a promising technology.


Long non coding RNA NRON inhibited breast cancer development through regulating miR-302b/SRSF2 axis.
Long noncoding RNA NRON has been investigated in various tumors, such as hepatocellular carcinoma. However, the role of lncRNA NRON in breast cancer remains unclear. The aim of this study was to explore the function and mechanism of lncRNA NRON in breast cancer. Overexpression and knockdown vectors were constructed. Proliferation and invasion were measured to evaluate the function of lncRNA NRON. A dual-luciferase reporter assay was utilized to analyze the potential binding target of lncRNA NRON. A rescue experiment was performed to verify the relationship between lncRNA NRON and SRSF2. Our results showed that the expression of lncRNA NRON was significantly downregulated in breast cancer tissues. Overexpression of lncRNA NRON significantly inhibited proliferation and invasion in breast cancer cell lines. Knockdown of lncRNA NRON promoted breast cancer development. We also provided evidence that lncRNA NRON negatively regulated miR-302b. Moreover, we identified SRSF2 as a downstream target of miR-302b. Overall, we performed a comprehensive analysis to indicate that the lncRNA NRON/miR-302b/SRSF2 axis plays an important role in breast cancer. Our study is the first to prove that lncRNA NRON functions as a tumor suppressor in breast cancer.


Innate immune surveillance of spontaneous B cell lymphomas by natural killer cells and gammadelta T cells.
Few studies have demonstrated that innate lymphocytes play a major role in preventing spontaneous tumor formation. We evaluated the development of spontaneous tumors in mice lacking beta-2 microglobulin (beta2m; and thus MHC class I, CD1d, and CD16) and/or perforin, since these tumor cells would be expected to activate innate effector cells. Approximately half the cohort of perforin gene-targeted mice succumbed to spontaneous disseminated B cell lymphomas and in mice that also lacked beta2m, the lymphomas developed earlier (by more than 100 d) and with greater incidence (84%). B cell lymphomas from perforin/beta2m gene-targeted mice effectively primed cell-mediated cytotoxicity and perforin, but not IFN-gamma, IL-12, or IL-18, was absolutely essential for tumor rejection. Activated NK1.1+ and gammadeltaTCR+ T cells were abundant at the tumor site, and transplanted tumors were strongly rejected by either, or both, of these cell types. Blockade of a number of different known costimulatory pathways failed to prevent tumor rejection. These results reflect a critical role for NK cells and gammadeltaTCR+ T cells in innate immune surveillance of B cell lymphomas, mediated by as yet undetermined pathway(s) of tumor recognition.


Bmi-1-RING1B prevents GATA4-dependent senescence-associated pathological cardiac hypertrophy by promoting autophagic degradation of GATA4.
Senescence-associated pathological cardiac hypertrophy (SA-PCH) is associated with upregulation of foetal genes, fibrosis, senescence-associated secretory phenotype (SASP), cardiac dysfunction and increased morbidity and mortality. Therefore, we conducted experiments to investigate whether GATA4 accumulation induces SA-PCH, and whether Bmi-1-RING1B promotes GATA4 ubiquitination and its selective autophagic degradation to prevent SA-PCH. Bmi-1-deficient (Bmi-1-/- ), transgenic Bmi-1 overexpressing (Bmi-1Tg ) and wild-type (WT) mice were infused with angiotensin II (Ang II) to stimulate the development of SA-PCH. Through bioinformatics analysis with RNA sequencing data from cardiac tissues, we found that Bmi-1-RING1B and autophagy are negatively related to SA-PCH. Bmi-1 deficiency promoted GATA4-dependent SA-PCH by increasing GATA4 protein and hypertrophy-related molecules transcribed by GATA4 such as ANP and BNP. Bmi-1 deficiency stimulated NF-κB-p65-dependent SASP, leading to cardiac dysfunction, cardiomyocyte hypertrophy and senescence. Bmi-1 overexpression repressed GATA4-dependent SA-PCH. GATA4 degraded by Bmi-1 was mainly dependent on autophagy rather than proteasome. In human myocardium, p16 positively correlated with ANP and GATA4 and negatively correlated with LC3B, Bmi-1 and RING1B; GATA4 positively correlated with p62 and negatively correlated with Bmi-1 and LC3B. With increased p16 protein levels, ANP-, BNP- and GATA4-positive cells or areas increased; however, LC3B-positive cells or areas decreased in human myocardium. GATA4 is ubiquitinated after combining with Bmi-1-RING1B, which is then recognised by p62, is translocated to autophagosomes to form autophagolysosomes and degraded. Downregulated GATA4 ameliorated SA-PCH and cardiac dysfunction by reducing GATA4-dependent hypertrophy and SASP-related molecules. Bmi-1 combined with RING1B (residues 1-179) and C-terminus of GATA4 (residues 206-443 including zinc finger domains) through residues 1-95, including a RING-HC-finger. RING1B combined with C-terminus of GATA4 through the C-terminus (residues 180-336). Adeno-associated viral vector serotype 9 (AAV9)-cytomegalovirus (CMV)-Bmi-1-RING1B treatment significantly attenuated GATA4-dependent SA-PCH through promoting GATA4 autophagic degradation. Bmi-1-RING1B maintained cardiac function and prevented SA-PCH by promoting selective autophagy for degrading GATA4. AAV9-CMV-Bmi-1-RING1B could be used for translational gene therapy to ubiquitinate GATA4 and prevent GATA4-dependent SA-PCH. Also, the combined domains between Bmi-1-RING1B and GATA4 in aging cardiomyocytes could be therapeutic targets for identifying stapled peptides in clinical applications to promote the combination of Bmi-1-RING1B with GATA4 and the ubiquitination of GATA4 to prevent SA-PCH and heart failure. We found that degradation of cardiac GATA4 by Bmi-1 was mainly dependent on autophagy rather than proteasome, and autophagy agonists metformin and rapamycin could ameliorate the SA-PCH, suggesting that activation of autophagy with metformin or rapamycin could also be a promising method to prevent SA-PCH.


Cyclic nucleotides levels in the liver of rats treated with CCl4.
Treatment of rats with two different doses of CCl4 (respectively 2.5 and 0.5 ml/kg body wt. intragastrically) is followed by a rapid increase in the cAMP content of the liver. With 0.5 ml of CCl4, the increase occurs as early as 30 min after poisoning, namely about 4-5 h before the onset of triglyceride accumulation in the liver. The maximum increase has been at 6 h after administration of the hepatotoxin. In both experimental conditions, normal values are recovered only after 36-48 h. cGMP level appears unmodified during the whole observation period. Therefore the ratio cGMP/CAMP decreases consistently. The ATP level decreases significantly between 2 and 12 h. The increase in liver triglycerides level after CCl4 can be also a consequence of an impairment of microtubule function, leading to a decreased release of lipoprotein micelles from hepatocytes into the blood stream.


The use of degrees of certainty to evaluate knowledge.
In patients with chronic diseases education should improve knowledge about the disease and increase certainty in knowledge. We present here a technique to measure changes in certainty after an educational intervention. For this purpose, before and after a course, patients answer a questionnaire in which answers are accompanied by an estimate of the degree of certainty. Answers are then assigned to areas of knowledge defined a priori: mastered (certainty > or = 90%, correctness > or = 90%), hazardous (certainty > or = 90%, correctness < or = 50%), uncertain (certainty < or = 50%, correctness > or = 90%) and residual. Finally differences in the distribution of answers among different areas are analysed statistically. Using this technique in a group of patients with type I diabetes who followed a course on insulin use, we found significant changes in the distribution of answers among different areas of knowledge. Thus changes in certainty can be analysed quantitatively and used to evaluate better the effect of therapeutic education.


Familiarity increases the number of remembered Pokémon in visual short-term memory.
Long-term memory (LTM) can influence many aspects of short-term memory (STM), including increased STM span. However, it is unclear whether LTM enhances the quantitative or qualitative aspect of STM. That is, do we retain a larger number of representations or more precise representations in STM for familiar stimuli than unfamiliar stimuli? This study took advantage of participants' prior rich multimedia experience with Pokémon, without investing on laboratory training to examine how prior LTM influenced visual STM. In a Pokémon visual STM change detection task, participants remembered more first-generation Pokémon characters that they were more familiar with than recent-generation Pokémon characters that they were less familiar with. No significant difference in memory quality was found when quantitative and qualitative effects of LTM were isolated using receiver operating characteristic (ROC) analyses. Critically, these effects were absent in participants who were unfamiliar with first-generation Pokémon. Furthermore, several alternative interpretations were ruled out, including general video-gaming experience, subjective Pokémon preference, and verbal encoding. Together, these results demonstrated a strong link between prior stimulus familiarity in LTM and visual STM storage capacity.


Integrated quality control: implementation and validation of instrument function checks and procedural controls for a cartridge-based point-of-care system for critical care analysis.
In this article, the process used to develop and validate an integrated quality-control system for a cartridge-based, point-of-care system for critical care analysis is outlined. Application of risk management principles has resulted in a quality control system using a combination of statistical quality control with onboard reference solutions and failure pattern recognition used to flag common failure modes during the analytical phase of the testing process. A combination of traditional external quality control, integrated quality control to monitor ongoing instrument functionality, operator training, and other laboratory-implemented monitors is most effective in controlling known failure modes during the testing process.


MSH receptor expression and the relationship to melanogenesis and metastatic activity in B16 melanoma.
In this study we have compared the effects of different pro-opiomelanocortin (POMC) peptides on melanogenesis and metastasis and their relationship to MSH receptor expression in B16F1 melanoma cells. All peptides, apart from beta-endorphin, increased melanogenesis and the order of potency was Nle4DPhe7-alpha-MSH greater than alpha-MSH greater than ACTH[1-39] greater than des-acetyl alpha-MSH greater than ACTH[1-24]. A similar order of potency was found for metastasis, except for ACTH [1-24], which had a relatively greater effect on metastasis. These findings suggest that the effects on melanogenesis and metastasis are mediated via the same receptor. The results of ligand binding studies also indicated the presence of a single receptor with a KD value for Nle4DPhe7-alpha-MSH of 62 +/- 16pM. This was consistent with crosslinking studies using [125I] Nle4DPhe7-alpha-MSH which produced a single 50-55 kD band on analysis by SDS-PAGE. However, the relative binding affinities of the different peptides, measured by displacement of [125I]-Nle4DPhe7-alpha-MSH, did not closely correlate with the relative potencies in stimulating melanogenesis and metastasis. This suggests that receptor activation and the subsequent biological response is not determined solely by binding affinity.


Influence of prolonged storage process, pasteurization, and heat treatment on biologically-active human milk proteins.
The bioactive proteins in human milk may be influenced by prolonged storage process, pasteurization, and heat treatment. This study was conducted to evaluate the effects of these procedures. Three forms of human milk - freshly expressed, frozen at -20°C for a prolonged duration, and pasteurized milk - were collected from 14 healthy lactating mothers and a milk bank. The concentrations of major bioactive proteins (secretory immunoglobulin A, lactoferrin, lysozyme, and leptin) were quantified using enzyme-linked immunosorbent assay kits. Changes in these proteins by heat treatment at 40°C or 60°C for 30 minutes were further evaluated. The mean concentrations of lactoferrin and secretory immunoglobulin A were significantly reduced by 66% and 25.9%, respectively, in pasteurized milk compared with those in freshly-expressed milk. Heat treatment at 40°C or 60°C did not cause significant changes in lactoferrin and secretory immunoglobulin A, but there was an apparent increase in lysozyme (p = 0.016). There were no significant differences in leptin level among these three forms of milk prior to (p = 0.153) or after heat treatment (p = 0.053). Various freezing/heating/pasteurization processes applied to human milk prior to delivery to neonates could affect the concentration of immunomodulatory proteins, especially lactoferrin, secretory immunoglobulin A, and lysozyme. Leptin was unaffected by the various handling processes tested. Fresh milk was found to be the best food for neonates. Further studies are warranted to evaluate the functional activity of these proteins and their effects on infants' immunological status.


Lipoaugmentation of the vocal folds: a survey on alternative donor sites for graft harvesting.
Lipoaugmentation is a treatment option for patients suffering from glottic insufficiency. Autologous fat is a nearly ideal material for vocal-fold augmentation from the view of biocompatibility and viscoelasticity, but there is still the problem of high graft resorption. As distribution and biological behavior of fatty tissue is very different in the human body, the aim of the study was to elucidate possible donor sites with respect to the quantity of harvested fat, the surgical accessibility to the region, the donor site morbidity and possibility of aesthetic defects and the quality of harvested tissue. Possible donor sites for harvesting were examined by magnetic resonance imaging in thirty-five patients with special emphasis to the buccal fat pad, the neck, the dorsolateral side of the proximal upper extremity, the subcutaneous layer of the abdominal wall, the superficial trochanteric region, the medial thigh, and the infrapatellar fat pad. Identified regions that failed to be chosen into consideration because of an elaborate surgical approach (superficial axillary's space, ischio-anal fossa, subcutaneous layer of buttock, popliteal fossa) were not taken into consideration. The mean volume of the buccal fat was 3.994 cm(3); the average thickness of the fat at the level of C7 was 1.721 cm, the mean value in the upper extremities was 1.913 cm laterally and 1.275 cm dorsally. The subcutaneous fat of the abdominal wall was divided into a superficial compartment (mean: 1.527 cm) and a deep one (average: 3.545 cm). In the superficial trochanteric region, the mean thickness was 2.536 cm, in the medial thigh 2.127 cm; the mean volume of the infrapatellar fat pad was 20.198 cm(3). All regions of interest showed reproducible and sufficient amounts of harvestable tissue, we found significant intersexual differences in dorsolateral side of the upper arm, subcutaneous layer of the abdominal wall and superficial trochanteric region. When harvesting subcutaneous tissue of the abdominal wall, grafts of the deep layer should be preferred, in the upper extremity the deep, muscle-neighbored parts. An alternative method is the surgically accessible fat of the neck. Solid fat pads could be harvested from the buccal region or the infrapatellar fat.


A cross-media content analysis of motivational themes in direct-to-consumer prescription drug advertising.
Direct-to-consumer (DTC) advertising of prescription drugs is a widely discussed issue in health care. However, little is known about the characteristics of the motivational themes used in this type of advertising. The aim of this study was to investigate the concurrent presentation of motivational themes in DTC print and television advertisements. The content analyses focused on advertisements of 2 targeted drug classes (cyclooxygenase-2 enzyme inhibitors and 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitors) in magazines and on television. Targeted print advertisements (for celecoxib, rofecoxib, atorvastatin, pravastatin, and simvastatin) from September to December 2001 and targeted television advertisements (for celecoxib, rofecoxib, and simvastatin) from November 2001 were investigated. The motivational themes were assessed using a theoretical framework based on self-regulatory focus theory and cultural orientation. Self-regulatory focus was examined in terms of goal orientation (promotion vs prevention) and emotional aspects, (e.g., cheerfulness, dejection, quiescence, agitation). The cultural orientation was examined in terms of individualism versus collectivism. The visual-verbal match was categorized as direct if the audio and visual information was semantically redundant, as partial if it was partially related, and as no match at all if it was different or conflicting. Twelve print advertisements in 10 magazines and 4 television advertisements on 4 television networks were examined; the interrater reliability scores from 3 independent, trained judges ranged from 0.93 to 0.99. The score was low (0.57) in the visual-verbal match measurement for television advertisements. Products in the same category appeared to be promoted using different self-regulatory foci. For example, celecoxib and atorvastatin advertisements tended to be promotion oriented, whereas pravastatin advertisements tended to be prevention oriented. Motivational themes were found throughout the print advertisements (e.g., pictures, headlines, main text). Only a few advertisements illustrated factual information about a product in a pictorial format. The cultural orientation of the advertisements was similar across brands, with individualistic appeals being common. On television, visual-verbal matches (either direct or partial matches) were generally found, except in the section where risk information was presented. In this content analysis, prescription drugs in the same class appeared to be promoted using different self-regulatory foci, but individualistic appeals were found more often than collectivistic appeals across brands.


The haemophilic patient's self-perception of changes in health and life-style arising from self-treatment.
Twenty three adults and twenty children with haemophilia, all on self-treatment, participated in the present study. They were interviewed as to how they perceive the effects of self-treatment upon various aspects of their life. It was found that the ability to treat themselves was perceived by patients as a great improvement over their previous treatment regimes for which they had had to go to hospital. Twelve of the adult patients had participated in our previous studies concerned with the social an psychological aspects of haemophilia. The analysis of the responses of these twelve patients showed that self-treatment is perceived so positively that some of the patients' earlier perceptions were distorted to correspond with their present and more optimistic perceptions of their condition. However, the number and type of psychosomatic symptoms was found to be unchanged by self-treatment and the unemployment rate has not decreased.


Immunolocalization of C1-tetrahydrofolate synthase in the rat kidney.
The distribution of the trifunctional enzyme C1-tetrahydrofolate synthase (C1-THF synthase) was examined in the rat kidney by immunolocalization with anti-C1-THF synthase serum using the peroxidase-antiperoxidase method. C1-THF synthase immunoreactivity was detected in both distal and proximal epithelial cells. Staining of the distal tubule epithelia was more intense and granular whereas staining of the proximal tubule epithelia was diffuse. All cells of the cortical collecting duct showed positive granular staining. In the outer medullary collecting duct, the intercalated cells showed intense granular cytoplasmic staining and the principal cells were either negative or weakly positive. The ascending thick limb of Henle's loop was also positive. Glomeruli and the inner medulla showed no staining for C1-THF synthase.


Cs(+) causes a voltage-dependent block of inward K currents in resting skeletal muscle fibres.
When frog skeletal muscle fibres are bathed in solutions containing Cs(+) and K(+) in the ratio 1:4,000, a reduction is observed in the size of inward K currents through the resting membrane. This effect is enhanced by an increase in either hyperpolarisation or external Cs(+) concentration. It can be predicted from these findings that regenerative changes in membrane potential should be obtainable in fibres, in the presence of Cs(+), that are hyperpolarised by means of a current electrode. Such responses are described in the last part of this report. In squid axon and frog node, internal Cs(+) produces a voltage-dependent block of the delayed, outward K currents, though the ratio of Cs(+) to K(+) required for this effect is far greater than that used in the experiments reported here. A closer parallel can be drawn between our findings and those recently reported on the inward K currents in the starfish egg cell.


Management of attention-deficit disorder and attention-deficit/hyperactivity disorder drug intoxication in dogs and cats.
Two types of drugs are generally used for treating attention-deficit/hyperactivity disorder or attention-deficit disorder in humans: amphetamines or similar stimulants and the nonamphetamine atomoxetine. We describe the toxicity and treatment of both amphetamines and similar medications and atomoxetine in dogs and cats. Amphetamine intoxication can cause life-threatening stimulatory signs. Treatment is aimed at preventing absorption, controlling the stimulatory signs, and protecting the kidneys; prognosis is generally good. Atomoxetine also has a fast onset of action; stimulatory signs such as hyperactivity and tachycardia are often seen. There are little published data about treatment of atomoxetine toxicity in cats and dogs.


Phenomenological research among Canadian and United States indigenous populations: oral tradition and quintessence of time.
Researchers conducting phenomenological studies among indigenous peoples in the United States and Canada have identified a seamless link between phenomenology and indigenous oral tradition. Phenomenology is compatible with indigenous peoples, because it is synchronous with holistic indigenous cultural lifeway and values. Phenomenology, as a research method, assists indigenous people in reproducing, through narrative communication, features of the past, present, and future. In the narrative process, this method elicits significant implicit meaning of indigenous culture and assists with recording the essence of experiences and events of indigenous societies. A product of the telling of narrative stories is the capacity to reflect on change that will enhance health in a holistic and culturally acceptable manner.


Probe signal correction for differential methylation hybridization experiments.
Non-biological signal (or noise) has been the bane of microarray analysis. Hybridization effects related to probe-sequence composition and DNA dye-probe interactions have been observed in differential methylation hybridization (DMH) microarray experiments as well as other effects inherent to the DMH protocol. We suggest two models to correct for non-biologically relevant probe signal with an overarching focus on probe-sequence composition. The estimated effects are evaluated and the strengths of the models are considered in the context of DMH analyses. The majority of estimated parameters were statistically significant in all considered models. Model selection for signal correction is based on interpretation of the estimated values and their biological significance.


Dietary Supplement Use among U.S. Children by Family Income, Food Security Level, and Nutrition Assistance Program Participation Status in 2011⁻2014.
This analysis characterizes use of dietary supplements (DS) and motivations for DS use among U.S. children (≤18 years) by family income level, food security status, and federal nutrition assistance program participation using the 2011⁻2014 National Health and Nutrition Examination Survey data. About one-third (32%) of children used DS, mostly multivitamin-minerals (MVM; 24%). DS and MVM use were associated with higher family income and higher household food security level. DS use was lowest among children in households participating in the Supplemental Nutrition Assistance Program (SNAP; 20%) and those participating in the Special Supplemental Nutrition Assistance Program for Women, Infants, and Children (WIC; 26%) compared to both income-eligible and income-ineligible nonparticipants. Most children who used DS took only one (83%) or two (12%) products; although children in low-income families took fewer products than those in higher income families. The most common motivations for DS and MVM use were to "improve (42% or 46%)" or "maintain (34 or 38%)" health, followed by "to supplement the diet (23 or 24%)" for DS or MVM, respectively. High-income children were more likely to use DS and MVM "to supplement the diet" than middle- or low-income children. Only 18% of child DS users took DS based on a health practitioner's recommendation. In conclusion, DS use was lower among children who were in low-income or food-insecure families, or families participating in nutrition assistance programs.


Proximity-dependent biotin identification links cholesterol catabolism with branched-chain amino acid degradation in Mycobacterium smegmatis.
Cholesterol is a crucial component in Mycobacterium tuberculosis virulence as it is required for phagocytosis of mycobacteria by macrophages. In addition, the tubercle bacilli can grow using cholesterol as the sole carbon source. Thus, cholesterol catabolism represents a valuable target for the development of new antitubercular drugs. However, the molecular partners of cholesterol catabolism remain elusive in mycobacteria. Here, we focused on HsaC and HsaD, enzymes involved in two consecutive steps of cholesterol ring degradation and identified putative partners, using a BirA-based proximity-dependent biotin identification (BioID) approach in Mycobacterium smegmatis. In rich medium, the fusion protein BirA-HsaD was able to fish the endogenous cognate HsaC, thus validating this approach to study protein-protein interactions and to infer metabolic channeling of cholesterol ring degradation. In chemically defined medium, both HsaC and HsaD interacted with four proteins, BkdA, BkdB, BkdC, and MSMEG_1634. BkdA, BkdB, and BkdC are enzymes that participate in the degradation of branched-chain amino acids. As cholesterol and branched-chain amino acid catabolism both generate propionyl-CoA, which is a toxic metabolite for mycobacteria, this interconnection suggests a compartmentalization to avoid dissemination of propionyl-CoA into the mycobacterial cytosol. Moreover, the BioID approach allowed us to decipher the interactome of MSMEG_1634 and MSMEG_6518, two proteins of unknown function, which are proximal to the enzymes involved in cholesterol and branched-chain amino acid catabolism. In conclusion, BioID is a powerful tool to characterize protein-protein interactions and to decipher the interconnections between different metabolic pathways, thereby facilitating the identification of new mycobacterial targets.


Physical and psychosocial correlates of head and neck cancer: an update of the literature and challenges for the future (1996-2003).
An update of the literature on physical and psychosocial aspects of head and neck cancer, with special emphasis on effects of treatment, patient-related factors and psychosocial intervention on quality of life (QoL). QoL deteriorates during and directly after treatment and returns only slowly to pre-treatment values. Organ preservation, gender and coping-strategies are factors related to QoL after treatment. As prognosis and survival were found to be comparable under different treatment regimes, we noticed a gradual shift in therapy towards organ preservation. Systematic care, using specific instruments is important for improving the QoL. We feel that more attention should be given to improving support, in order to optimize the QoL of patients during the palliative stage.


Preaxial polydactyly associated with a MSX1 mutation and report of two novel mutations.
We report two novel heterozygous missense MSX1 mutations in two Thai families (c.739C>T; p.Pro247Ser and c.607G>A; p.Ala203Thr). The p.Ala203Thr mutation was found in a female patient, her sister, and their father and is associated with unilateral cleft lip and palate, hypodontia, and microdontia. The p.Pro247Ser mutation was found in a three-generation Thai family and was associated with bilateral cleft lip and palate, hypodontia, microdontia, and dens invaginatus. The proband also had preaxial polydactyly of the left hand. The role of Msx1 in limb development in mice is discussed. Intrafamilial variability of the phenotypes is clearly evident. This is the first time that a limb anomaly has been reported to be associated with a mutation in MSX1.


A potential propensity for failure secondary to clot embolism in neonatal ECMO.
To report a single case of oxygenator failure caused by clot embolism originating from the bladder; and to discuss some preventative options. A 2.5 kg neonate with a diagnosis of influenza A received veno-arterial (V-A) extracorporeal membrane oxygenation (ECMO) for cardiorespiratory support. Halfway through treatment, she underwent an elective circuit change for numerous clots in her circuit. The patient continued to consume vast quantities of platelets and developed a fatal oxygenator failure after 18 days. Amongst the factors influencing the outcome in events of a sudden unexpected oxygenator failure are the severity of patient illness, the size of the clot relative to the size of the oxygenator, the availability of a previously primed circuit and the ease and speed of priming a new oxygenator. There is a need for improvement in the design of small oxygenators and ECMO circuits. Adjustment of the coagulation parameters and lowering the tolerance towards clots in the circuit by electively changing them may reduce the incidence of sudden unexpected oxygenator failure. However, using a slightly larger Medos oxygenator may gain valuable time needed to arrange an oxygenator/circuit change.


Context effects on tempo and pleasantness judgments for Beatles songs.
Context effects on tempo and pleasantness judgments of different tempos were demonstrated in three experiments using Beatles songs. In Experiments 1 and 2, we explored how listening to versions of the same song that were played at different tempos affected tempo and pleasantness ratings. In both experiments, contrast effects were found on judgments of tempo, with target tempos rated faster when context tempos were slow than when they were fast. In both experiments, we also showed that the peak of the pleasantness rating function shifted toward the values of the context tempos, reflecting disordinal context effects on pleasantness relationships. Familiarity with the songs did not moderate these effects, and shifts in tempo ratings did not correlate with shifts in most pleasant target tempos when context was manipulated within subjects. In Experiment 3, we examined how manipulations of context tempos for one song affected judgments of the same song as compared with judgments of other more or less similar songs. For tempo ratings, contrast effects transferred to ratings of a similar song, but for pleasantness ratings, assimilative shifts of ideals were found only for the same song and not for similar songs. This pattern of results was supportive of independent bases for the two context effects.


CPEB4-CLOCK crosstalk during temporal lobe epilepsy.
Posttranscriptional mechanisms are increasingly recognized as important contributors to the formation of hyperexcitable networks in epilepsy. Messenger RNA (mRNA) polyadenylation is a key regulatory mechanism governing protein expression by enhancing mRNA stability and translation. Previous studies have shown large-scale changes in mRNA polyadenylation in the hippocampus of mice during epilepsy development. The cytoplasmic polyadenylation element-binding protein CPEB4 was found to drive epilepsy-induced poly(A) tail changes, and mice lacking CPEB4 develop a more severe seizure and epilepsy phenotype. The mechanisms controlling CPEB4 function and the downstream pathways that influence the recurrence of spontaneous seizures in epilepsy remain poorly understood. Status epilepticus was induced in wild-type and CPEB4-deficient male mice via an intra-amygdala microinjection of kainic acid. CLOCK binding to the CPEB4 promoter was analyzed via chromatin immunoprecipitation assay and melatonin levels via high-performance liquid chromatography in plasma. Here, we show increased binding of CLOCK to recognition sites in the CPEB4 promoter region during status epilepticus in mice and increased Cpeb4 mRNA levels in N2A cells overexpressing CLOCK. Bioinformatic analysis of CPEB4-dependent genes undergoing changes in their poly(A) tail during epilepsy found that genes involved in the regulation of circadian rhythms are particularly enriched. Clock transcripts displayed a longer poly(A) tail length in the hippocampus of mice post-status epilepticus and during epilepsy. Moreover, CLOCK expression was increased in the hippocampus in mice post-status epilepticus and during epilepsy, and in resected hippocampus and cortex of patients with drug-resistant temporal lobe epilepsy. Furthermore, CPEB4 is required for CLOCK expression after status epilepticus, with lower levels in CPEB4-deficient compared to wild-type mice. Last, CPEB4-deficient mice showed altered circadian function, including altered melatonin blood levels and altered clustering of spontaneous seizures during the day. Our results reveal a new positive transcriptional-translational feedback loop involving CPEB4 and CLOCK, which may contribute to the regulation of the sleep-wake cycle during epilepsy.


How Foot Progression Angle Affects Knee Adduction Moment and Angular Impulse in Patients With and Without Medial Knee Osteoarthritis: A Meta-Analysis.
To investigate effects of foot progression angle (FPA) modification on the first and second peaks of external knee adduction moment (EKAM) and knee adduction angular impulse (KAAI) in individuals with and without medial knee osteoarthritis (OA) during level walking. PubMed, Embase, the Cumulative Index to Nursing and Allied Health Literature (CINAHL), Web of Science, and SPORTDiscus were searched from inception to February 2020 by 2 independent reviewers. Included studies compared FPA modification (toe-in or toe-out gait) interventions to lower EKAM and/or KAAI with natural walking. Studies were required to report the first or second peaks of EKAM or KAAI. Sixteen studies were included, and >85% of included patients were graded with Kellgren/Lawrence grade II-IV knee OA. Toe-in gait reduced the first EKAM peak (standardized mean difference [SMD] -0.75 [95% confidence interval (95% CI) -1.05, -0.45]) and KAAI (SMD -0.46 [95% CI -0.86, -0.07]), while toe-out gait reduced the second EKAM peak (SMD -1.04 [95% CI -1.34, -0.75]) in healthy individuals. For patients with knee OA, toe-out gait reduced the second EKAM peak (SMD -0.53 [95% CI -0.75, -0.31]) and KAAI (SMD -0.26 [95% CI -0.49, -0.03]), while toe-in gait did not affect both EKAM peaks and KAAI. Discrepancy in biomechanical effects of FPA modification was demonstrated between individuals with and without medial knee OA. Compared with natural walking, both toe-in and toe-out gait may be more effective in lowering EKAM and KAAI in healthy individuals. Toe-out gait may reduce EKAM and KAAI in patients with mild-to-severe knee OA. There is insufficient data from patients with early-stage knee OA, indicating that future research is required.


Gluteal muscle and subcutaneous tissue thicknesses in adults: a systematic review and meta-analysis.
Nurses often administer intramuscular injections at the gluteal site. This study aimed to determine gluteal muscle and subcutaneous tissue thicknesses in adults. Systematic review and meta-analysis. The databases Turkish Medline, Ulakbim, National Thesis Center, Cochrane, Web of Science, Science Direct, PubMed, CINAHL Plus with Full text (EBSCO host), OVID and SCOPUS were screened using the keywords 'intramuscular injection', 'subcutaneous tissue thickness', 'muscle tissue thickness' and 'needle length' between April and May 2021. The studies were evaluated with ultrasound. This study was reported according to the PRISMA recommendations. Six studies met the eligibility criteria. The total sample size was 734 (women: 432, men: 302). The V method revealed that the ventrogluteal site had a muscle and subcutaneous tissue thickness of 38.071 ± 2.119 and 19.927 ± 2.493 mm, respectively. The geometric method revealed that the ventrogluteal site had a muscle and subcutaneous tissue thickness of 35.989 ± 4.190 and 19.661 ± 3.992 mm, respectively. The geometric method also revealed that the dorsogluteal site had a thickness of 42.560 ± 8.840 mm. According to the V method, females had thicker subcutaneous tissue at the ventrogluteal site than males (Q = 5.37, df = 1, p = 0.0204). Body mass index did not affect the subcutaneous tissue thicknesses at the ventrogluteal site. The results show that gluteal muscle, subcutaneous and total tissue thicknesses vary across injection sites.


Differing rates of fungi in sinonasal cultures from pediatric sinusitis patients.
Pediatric chronic rhinosinusitis (PCRS) is a unique clinical entity and the underlying source of inflammation is unknown. Certain subgroups, such as children with nasal polyps and cystic fibrosis (CF) sinusitis are often recalcitrant to standard medical PCRS treatments that target bacterial inflammation. Fungal infection and allergy to fungal proteins drive inflammation in other airway diseases, resulting in chronic inflammation of both the upper and lower airways. However, there is limited understanding of the role of fungi in the pathophysiology of PCRS. The objective of this study is to define the frequency of fungal infection in pediatric CRS patients, hypothesizing that certain subgroups may have more frequent positive fungal sinus cultures than other subgroups of pediatric sinusitis. Retrospective study of patients undergoing sinus surgery at a tertiary care pediatric hospital to determine the period prevalence of positive fungal cultures in subgroups of patients. 400 children from 2012 to 2019 were included. 265 patients had surgical culture results available. Of the 52 patients with CF 11 (21%) had positive fungal sinus cultures. Similarly, 28% of the 25 patients with non-CF nasal polyps had positive cultures. Only 8.2% of 110 CRS without polyps patients had positive cultures, significantly fewer than other subgroups (X2 (1, N = 240) = 17.22, p < 0.01). Children with CF and children with nasal polyps had more frequent positive fungal cultures than children without nasal polyps having sinus surgery. This confirms that pediatric CF and pediatric CRS with polyps represent unique populations to study the impact of fungal infection in CRS. Further research is required to determine if these fungi represent colonization or contribute to the inflammatory environment of the airways.


Role of traditional healers in the management of severe malaria among children below five years of age: the case of Kilosa and Handeni Districts, Tanzania.
The current malaria control strategy of WHO centres on early diagnosis and prompt treatment using effective drugs. Children with severe malaria are often brought late to health facilities and traditional health practitioners are said to be the main cause of treatment delay. In the context of the Rectal Artesunate Project in Tanzania, the role of traditional healers in the management of severe malaria in children was studied. A community cross-sectional study was conducted in Kilosa and Handeni Districts, involving four villages selected on the basis of existing statistics on the number of traditional health practitioners involved in the management of severe malaria. A total of 41 traditional health practitioners were selected using the snowballing technique, whereby in-depth interviews were used to collect information. Eight Focus Group Discussions (FGDs) involving traditional health practitioners, caregivers and community leaders were carried out in each district. Home management of fever involving sponging or washing with warm water at the household level, was widely practiced by caregivers. One important finding was that traditional health practitioners and mothers were not linking the local illness termed degedege, a prominent feature in severe malaria, to biomedically-defined malaria. The majority of mothers (75%) considered degedege to be caused by evil spirits. The healing process was therefore organized in stages and failure to abide to the procedure could lead to relapse of degedege, which was believed to be caused by evil spirits. Treatment seeking was, therefore, a complex process and mothers would consult traditional health practitioners and modern health care providers, back and forth. Referrals to health facilities increased during the Rectal Artesunate Project, whereby project staff facilitated the process after traditional medical care with the provision of suppositories. This finding is challenging the common view that traditional healers are an important factor of delay for malaria treatment, they actually play a pivotal role by giving "bio-medically accepted first aid" which leads to reduction in body temperature hence increasing chances of survival for the child. Increasing the collaboration between traditional healers and modern health care providers was shown to improve the management of severe malaria in the studied areas. Traditional health care is not necessarily a significant impediment or a delaying factor in the treatment of severe malaria. There is a need to foster training on the management of severe cases, periodically involving both traditional health practitioners and health workers to identify modalities of better collaboration.


Perceived fidelity of compressed and reconstructed radiological images: a preliminary exploration of compression, luminance, and viewing distance.
The authors' goal was to explore the impact of image compression algorithm and ratio, image luminance, and viewing distance on radiologists' perception of reconstructed image fidelity. Five radiologists viewed 16 sets of four hard-copy chest radiographs prepared for secondary interpretation. Each set included one uncompressed, and three compressed and reconstructed images prepared using three different algorithms but the same compression ratio. The sets were prepared using two subjects, four compression ratios (10:1, 20:1, 30:1, 40:1), and two luminance levels (2,400 cd/m2, standard lightbox illumination, and 200 cd/m2, simulating a typical CRT display). Readers ranked image quality and evaluated obviousness and clinical importance of differences. Viewing distances for image screening, inspection, and comparison were recorded. At 10:1 compression, the compressed and uncompressed images were nearly indistinguishable; the three algorithms were very similar, and differences were rated "not obvious" and "not important." At higher compression, readers consistently preferred uncompressed images, with notable differences between algorithms. The obviousness and clinical importance of differences were rated higher at lightbox luminance. Viewing distances appeared to be idiosyncratic.


The effect of calcium ions on the responses to motoneurons to substance P and eledoisin-related peptide in the toad and rat spinal cord.
The effects of changes in the external calcium ion concentration [Ca2+]0 on segmental reflexes and the responses of motoneurons to bath applied agonists have been studied in the toad and rat spinal cords in vitro. Reducing [Ca2+]0 enhanced polysynaptic reflexes in the toad, with maximal discharges occurring at 0.3 mM. Monosynaptic reflexes in the rat were reduced by lowering [Ca2+]0. Responses of toad motoneurons to substance P and eledoisin-related peptide were enhanced by lowering [Ca2+]0, maximum responses occurring at 0.3 mM. Lowering [Ca2+]0 also enhanced responses to L-glutamate but the effect was smaller and less consistent. This effect of Ca2+ was abolished by the addition of tetrodotoxin to the bathing solution. Toad motoneuron responses to gamma-aminobutyrate and glycine were not affected by alterations in [Ca2+]0. Rat motoneuron responses to substance P and eledoisin-related peptide were also enhanced by reductions in [Ca2+]0 but the effect was more pronounced in younger (less than 5 days post partum) than older (5-10 days post partum) animals. These results are consistent with the idea that motoneuron responses to peptides in normal solutions result from activation of receptors on both motoneurons and interneurons: enhancement of the responses by lowering [Ca2+]0 results from the potentiation of the transynaptic component.


The dynamics of density dependent population models.
The dynamics of density-dependent population models can be extraordinarily complex as numerous authors have displayed in numerical simulations. Here we commence a theoretical analysis of the mathematical mechanisms underlying this complexity from the viewpoint of modern dynamical systems theory. After discussing the chaotic behavior of one-dimensional difference equations we proceed to illustrate the general theory on a density-dependent Leslie model with two age classes. The pattern of bofurcations away from the equilibrium point is investigated and the existence of a "strange attractor" is demonstrated--i.e. an attracting limit set which is neither an equilibrium nor a limit cycle. Near the strange attractor the system exhibits essentially random behavior. An approach to the statical analysis of the dynamics in the chaotic regime is suggested. We then generalize our conclusions to higher dimensions and continuous models (e.g. the nonlinear von Foerster equation).


Skullcapflavone I from Scutellaria baicalensis induces apoptosis in activated rat hepatic stellate cells.
The therapeutic goal in liver fibrosis is the reversal of fibrosis and the selective clearance by apoptosis of hepatic stellate cells (HSCs), which play a central role in liver fibrogenesis. In this study, the apoptotic effect of wogonin, oroxylin A, 2',5,6',7-tetrahydroxyflavone, skullcapflavone I, and baicalein, isolated from the dried root of Scutellaria baicalensis, was investigated in activated rat HSCs, T-HSC/Cl-6 cells transformed with the Simian virus 40. Among the isolated compounds, skullcapflavone I (20 microM for 24 h) significantly induced apoptosis in activated rat HSCs while there was no change in the cell viability of hepatocytes. Skullcapflavone I increased caspase-3 and -9 activities accompanied by the proteolytic cleavage of poly(ADP-ribose) polymerase. Specific inhibitors of caspase-3 and caspase-9 prevented the apoptotic process induced by skullcapflavone I. From these results, skullcapflavone I from S. baicalensis selectively induced apoptosis in T-HSC/Cl-6 cells via caspase-3 and caspase-9 activation.


Frailty Measures of Patient-reported Activity and Fatigue May Predict 1-year Outcomes in Ambulatory Advanced Heart Failure: A Report From the REVIVAL Registry.
The Fried Frailty Phenotype predicts adverse outcomes in geriatric populations, but has not been well-studied in advanced heart failure (HF). The Registry Evaluation of Vital Information for Ventricular Assist Devices (VADs) in Ambulatory Life (REVIVAL) study prospectively collected frailty measures in patients with advanced HF to determine relevant assessments and their impact on clinical outcomes. HF-Fried Frailty was defined by 5 baseline components (1 point each): (1) weakness: hand grip strength less than 25% of body weight; (2) slowness based on time to walk 15 feet; (3) weight loss of more than 10 lbs in the past year; (4) inactivity; and (5) exhaustion, both assessed by the Kansas City Cardiomyopathy Questionnaire. A score of 0 or 1 was deemed nonfrail, 2 prefrail, and 3 or greater was considered frail. The primary composite outcome was durable mechanical circulatory support implantation, cardiac transplant or death at 1 year. Event-free survival for each group was determined by the Kaplan-Meier method and the hazard of prefrailty and frailty were compared with nonfrailty with proportional hazards modeling. Among 345 patients with all 5 frailty domains assessed, frailty was present in 17%, prefrailty in 40%, and 43% were nonfrail, with 67% (n = 232) meeting the criteria based on inactivity and 54% (n = 186) for exhaustion. Frail patients had an increased risk of the primary composite outcome (unadjusted hazard ratio [HR] 2.82, 95% confidence interval [CI] 1.52-5.24; adjusted HR 3.41, 95% CI 1.79-6.52), as did prefrail patients (unadjusted HR 1.97, 95% CI 1.14-3.41; adjusted HR 2.11, 95% CI 1.21-3.66) compared with nonfrail patients, however, the predictive value of HF-Fried Frailty criteria was modest (Harrel's C-statistic of 0.603, P = .004). The HF-Fried Frailty criteria had only modest predictive power in identifying ambulatory patients with advanced HF at high risk for durable mechanical circulatory support, transplant, or death within 1 year, driven primarily by assessments of inactivity and exhaustion. Focus on these patient-reported measures may better inform clinical trajectories in this population.


A comparative study of R functions for clustered data analysis.
Clustered or correlated outcome data is common in medical research studies, such as the analysis of national or international disease registries, or cluster-randomized trials, where groups of trial participants, instead of each trial participant, are randomized to interventions. Within-group correlation in studies with clustered data requires the use of specific statistical methods, such as generalized estimating equations and mixed-effects models, to account for this correlation and support unbiased statistical inference. We compare different approaches to estimating generalized estimating equations and mixed effects models for a continuous outcome in R through a simulation study and a data example. The methods are implemented through four popular functions of the statistical software R, "geese", "gls", "lme", and "lmer". In the simulation study, we compare the mean squared error of estimating all the model parameters and compare the coverage proportion of the 95% confidence intervals. In the data analysis, we compare estimation of the intervention effect and the intra-class correlation. In the simulation study, the function "lme" takes the least computation time. There is no difference in the mean squared error of the four functions. The "lmer" function provides better coverage of the fixed effects when the number of clusters is small as 10. The function "gls" produces close to nominal scale confidence intervals of the intra-class correlation. In the data analysis and the "gls" function yields a positive estimate of the intra-class correlation while the "geese" function gives a negative estimate. Neither of the confidence intervals contains the value zero. The "gls" function efficiently produces an estimate of the intra-class correlation with a confidence interval. When the within-group correlation is as high as 0.5, the confidence interval is not always obtainable.


The phenothiazine-class antipsychotic drugs prochlorperazine and trifluoperazine are potent allosteric modulators of the human P2X7 receptor.
P2X7, an ATP-gated cation channel, is involved in immune cell activation, hyperalgesia and neuropathic pain. By regulating cytokine release in the brain, P2X7 has been linked to the pathophysiology of mood disorders and schizophrenia. We here assess the impact of 123 drugs that act in the central nervous system on human P2X7. Most prominently, the tricyclic antipsychotics prochlorperazine (PCP) and trifluoperazine (TFP) potently inhibited P2X7-mediated Ca2+ entry, dye permeation and ionic currents. In divalent cation-containing bath solutions or after prolonged incubation, ATP-evoked P2X7 currents were inhibited by 10 μM PCP. This effect was not related to dopamine receptor antagonism. Surprisingly, PCP co-applied with ATP enhanced inward currents in bath solutions with low divalent cation concentrations. Intracellular perfusion with PCP did not substitute for the extracellularly applied drug, indicating that its binding sites are accessible from the extracellular space. Since P2X7 current potentiation by PCP was voltage-dependent, at least one site may be located within the electrical field of the membrane. While the channel opening and closure kinetic was altered by PCP, the apparent affinity of ATP remained unchanged (potentiation) or changed slightly (inhibition). Measurements in human monocyte-derived macrophages confirmed the PCP-induced inhibition of ATP-evoked Ca2+ influx, Yo-Pro-1 permeability, and whole cell currents. Interestingly, neither heterologously expressed rat or mouse P2X7 nor native P2X7 in rat astrocyte cultures or in mouse bone marrow-derived macrophages were inhibited by perazines with a similar potency. We conclude that perazine-type neuroleptics are potent, but species-selective allosteric modulators of human but not murine P2X7 receptors.


Death of sensory ganglion neurons after acute withdrawal of nerve growth factor in dissociated cell cultures.
The time course of dependence on nerve growth factor (NGF) for survival in sensory neurons in vitro was examined with microscopic and biochemical methods. Primary dorsal root ganglion (DRG) cultures from embryonic-day-15 (E-15) and day-19 (E-19) rats were maintained with standard dissociated cell culture techniques in the absence of most non-neuronal cells. After various times in culture, neurons were acutely deprived of neurotrophic support by changing to NGF-free medium and adding NGF antiserum to eliminate any residual NGF. Neuronal cultures were examined with phase microscopy; and, their metabolic activity was measured with a protein assay at various time points after NGF deprivation. E-15 neurons grown in culture for 5 days were exquisitely sensitive to acute NGF deprivation. By 12 h after NGF deprivation, neuronal morphology was severely disrupted and the majority of neurons appeared dead. E-15 neurons grown in culture for 8 or 11 days showed progressively less dependence on NGF for survival. These older neurons did not die until 24 and 48 h, respectively, following NGF withdrawal. Neurons grown in culture for 20 days did not show any morphologic changes by phase microscopy up to 4 days after NGF deprivation. Protein incorporation progressively decreased between 12 and 48 h after NGF withdrawal in E-15 neurons grown in culture for 5, 8, or 11 days.(ABSTRACT TRUNCATED AT 250 WORDS)


Road network safety evaluation using Bayesian hierarchical joint model.
Safety and efficiency are commonly regarded as two significant performance indicators of transportation systems. In practice, road network planning has focused on road capacity and transport efficiency whereas the safety level of a road network has received little attention in the planning stage. This study develops a Bayesian hierarchical joint model for road network safety evaluation to help planners take traffic safety into account when planning a road network. The proposed model establishes relationships between road network risk and micro-level variables related to road entities and traffic volume, as well as socioeconomic, trip generation and network density variables at macro level which are generally used for long term transportation plans. In addition, network spatial correlation between intersections and their connected road segments is also considered in the model. A road network is elaborately selected in order to compare the proposed hierarchical joint model with a previous joint model and a negative binomial model. According to the results of the model comparison, the hierarchical joint model outperforms the joint model and negative binomial model in terms of the goodness-of-fit and predictive performance, which indicates the reasonableness of considering the hierarchical data structure in crash prediction and analysis. Moreover, both random effects at the TAZ level and the spatial correlation between intersections and their adjacent segments are found to be significant, supporting the employment of the hierarchical joint model as an alternative in road-network-level safety modeling as well.


A Reliability Comparison of Cone-Beam Breast Computed Tomography and Mammography: Breast Density Assessment Referring to the Fifth Edition of the BI-RADS Atlas.
To evaluate the reliability of cone-beam breast computed tomography (CBBCT) in visual assessment of breast density referring to the fifth edition of the Breast Imaging Reporting and Data System compared to digital mammography. Breast density assessments of 130 female patients were performed by five radiologists referring to the fifth edition of Breast Imaging Reporting and Data System atlas both on two-view mammograms and CBBCT images. Assessments were repeated by three radiologists with different experience more than 1 month after the initial evaluation. The inter- and intrareader agreements were compared by using the Cohen's weighted Kappa statistic and intraclass correlation coefficient. Weighted Kappa statistic was also used to analyze the agreement between CBBCT images and mammograms. The influence of radiologist experience for breast density assessment was analyzed using a chi-square test. For CBBCT images, the inter-reader agreement was 0.781, whereas the agreement on mammograms was 0.744, both demonstrating moderate agreement. The level of intrareader reliability was higher on the CBBCT images than mammograms for breast density evaluation, 0.856 versus 0.786. Based on the majority report, the agreement between these two modalities was on substantial agreement degree. There was a statistically significant difference among radiologists with different levels of experience, and higher density categories were reported more often by experienced reader. CBBCT showed equal aptitude and better agreement for the breast density evaluation compared to mammography. CBBCT could be an effective modality for breast density assessment and breast cancer risk evaluation in routine diagnosis and breast cancer screening.


Social Chemical Communication Determines Recovery From L1 Arrest via DAF-16 Activation.
In a population, chemical communication determines the response of animals to changing environmental conditions, what leads to an enhanced resistance against stressors. In response to starvation, the nematode Caenorhabditis elegans arrest post-embryonic development at the first larval stage (L1) right after hatching. As arrested L1 larvae, C. elegans become more resistant to diverse stresses, allowing them to survive for several weeks expecting to encounter more favorable conditions. L1 arrested at high densities display an enhanced resistance to starvation, dependent on soluble compounds released beyond hatching and the first day of arrest. Here, we show that this chemical communication also influences recovery after prolonged periods in L1 arrest. Animals at high density recovered faster than animals at low density. We found that the density effect on survival depends on the final effector of the insulin signaling pathway, the transcription factor DAF-16. Moreover, DAF-16 activation was higher at high density, consistent with a lower expression of the insulin-like peptide DAF-28 in the neurons. The improved recovery of animals after arrest at high density depended on soluble compounds present in the media of arrested L1s. In an effort to find the nature of these compounds, we investigated the disaccharide trehalose as putative signaling molecule, since its production is enhanced during L1 arrest and it is able to activate DAF-16. We detected the presence of trehalose in the medium of arrested L1 larvae at a low concentration. The addition of this concentration of trehalose to animals arrested at low density was enough to rescue DAF-28 production and DAF-16 activation to the levels of animals arrested at high density. However, despite activating DAF-16, trehalose was not capable of reversing survival and recovery phenotypes, suggesting the participation of additional signaling molecules. With all, here we describe a molecular mechanism underlying social communication that allows C. elegans to maintain arrested L1 larvae ready to quickly recover as soon as they encounter nutrient sources.


A general model for the solubilities of gases in liquids.
The solubility of a compound is one of its most important properties. Here, regression relationships are presented for solubilities of a series of gases in water and in four organic solvents, treating each solvent separately. The solubilities are related to the Coulombic intermolecular interactions arising from the intrinsic polarities of the solute molecules and the polarities induced in them by the solvent. As a measure of intrinsic polarity, a statistical quantity defined in terms of the solute's molecular electrostatic potential is used, and the measure of induced polarity is taken to be the solute's molecular polarizability. Regression analyses show that solubility in water is best expressed in terms of just the intrinsic polarities of the solutes, but for the organic solvents, it is necessary to take into account both the intrinsic and the induced polarities of the solutes. If the dielectric constant of the solvent is included in the regression analysis, then a single relationship can encompass all four organic solvents. Solute molecular volumes were not found to contribute significantly to the present relationships.


Newborn hearing screening in the NICU: profile of failed auditory brainstem response/passed otoacoustic emission.
Incidence of a specific pattern of auditory responses, absent auditory brainstem responses (ABRs) and present otoacoustic emissions (OAEs), in newborn hearing screening in a regional perinatal center neonatal intensive care unit (NICU) is described. This profile, labeled auditory neuropathy or auditory dyssynchrony (AN/AD), is a dysfunction in neural/brainstem transmission that occurs in individuals whose outer hairs cells are functioning normally. Although the AN/AD profile has been associated with various risk factors, incidence and prediction are unknown. Analysis of electrophysiologic measures and medical record reviews of the first 22 months of the universal newborn hearing-screening program was conducted. Association of the AN/AD profile was evaluated with the following factors: gender, gestational age, ototoxic drug regimen, low birth weight, hyperbilirubinemia, hydrocephalus, low Apgar score, anoxia, respiratory distress syndrome, pulmonary hypertension, intraventricular hemorrhage, multiple birth, seizure activity, and family history. One hundred fifteen (24.1%) of the 477 infants failed the ABR in 1 or both ears and passed OAEs bilaterally. Comparisons of infants fitting the AN/AD profile with those not fitting the AN/AD profile were negative with 3 exceptions: those with hyperbilirubinemia and those who were administered vancomycin or furosemide. A logistic-regression analysis model failed to predict which infants would be at risk for the AN/AD profile either unilaterally or bilaterally. Screening of NICU infants should be conducted with ABR first, followed by OAE after failure on ABR. Because the incidence of the AN/AD profile was found to be 24% in this at-risk population, additional study is warranted.


Development and clinical application of a preimplantation genetic testing for monogenic disease (PGT-M) for beta thalassemia in Vietnam.
The purpose of this research is to study the clinical outcomes using a next-generation sequencing-based protocol allowing for simultaneous testing of mutations in the beta thalassemia (HBB) gene, including single nucleotide polymorphism (SNP) markers for PGT-M along with low-pass whole genome analysis of chromosome aneuploidies for PGT-A. A combined PGT-M (thalassemia) plus PGT-A system was developed for patients undergoing IVF in Vietnam. Here we developed a system for testing numerous thalassemia mutations plus SNP-based testing for backup mutation analysis and contamination control using next-generation sequencing (NGS). Low -pass next-generation sequencing was used to assess aneuploidy in some of the clinical PGT cases. Patients underwent IVF followed by embryo biopsy at the blastocyst stage for combined PGT-A/M. Two cases have completed the entire process including transfer of embryos, while a further nine cases have completed the IVF and PGT-M/A analysis but have not completed embryo transfer. In the two cases with embryo transfer, both patients achieved pregnancy with an unaffected, euploid embryo confirmed through prenatal diagnosis. In the further nine cases, 39 embryos were biopsied and all passed QC for amplification. There were 8 unaffected embryos, 31 carrier embryos, and 11 affected embryos. A subset of 24 embryos also had PGT-A analysis with 22 euploid embryos and 2 aneuploid embryos. Here we report the development and clinical application of a combined PGT-M for HBB and PGT-A for gross chromosome aneuploidies from 11 patients with detailed laboratory findings along with 2 cases that have completed embryo transfer.


Prognostic significance of the presence of intraductal carcinoma of the prostate and bone metastasis in needle biopsy for prostate carcinoma patients with Grade Group 5.
Intraductal carcinoma of the prostate (IDC-P) and bone metastasis have been both identified to associate with unfavorable clinical outcome of the prostate carcinoma (PCa). Our objective is to examine whether IDC-P or bone metastasis at diagnostic biopsies was associated with each other and whether they were linked with overall survival (OS) and cancer specific survival (CSS) of Grade Group 5 patients. We retrospectively selected the prostate biopsy specimens of 120 PCa patients with Grade Group 5 from Qilu Hospital of Shandong University between 2012 and 2016. There were 12 patients with IDC-P only, 52 patients with bone metastasis only and 10 patients with both IDC-P and bone metastasis. Overall, there was a significant correlation between the presences of the IDC-P and bone metastasis (P = 0.003). Kaplan-Meier survival analysis demonstrated that the presence of IDC-P and bone metastasis in diagnostic needle biopsy both conferred unfavorable CSS of Grade Group 5 patients. In addition, the presence of bone metastasis was a poor predictor of OS. Univariate and multivariate analysis revealed that bone metastasis was an independent prognostic factor for OS of Grade Group 5 patients, but IDC-P failed to be significant for either OS or CSS. Collectively, our study suggested that bone metastasis is an important prognostic factor and superior than the presence of the IDC-P for PCa patients with Grade Group 5.


Grapevine trunk diseases under thermal and water stresses.
Heat and water stresses, individually or combined, affect both the plant (development, physiology, and production) and the pathogens (growth, morphology, dissemination, distribution, and virulence). The grapevine response to combined abiotic and biotic stresses is complex and cannot be inferred from the response to each single stress. Several factors might impact the response and the recovery of the grapevine, such as the intensity, duration, and timing of the stresses. In the heat/water stress-GTDs-grapevine interaction, the nature of the pathogens, and the host, i.e., the nature of the rootstock, the cultivar and the clone, has a great importance. This review highlights the lack of studies investigating the response to combined stresses, in particular molecular studies, and the misreading of the relationship between rootstock and scion in the relationship GTDs/abiotic stresses. Grapevine trunk diseases (GTDs) are one of the biggest threats to vineyard sustainability in the next 30 years. Although many treatments and practices are used to manage GTDs, there has been an increase in the prevalence of these diseases due to several factors such as vineyard intensification, aging vineyards, or nursery practices. The ban of efficient treatments, i.e., sodium arsenite, carbendazim, and benomyl, in the early 2000s may be partly responsible for the fast spread of these diseases. However, GTD-associated fungi can act as endophytes for several years on, or inside the vine until the appearance of the first symptoms. This prompted several researchers to hypothesise that abiotic conditions, especially thermal and water stresses, were involved in the initiation of GTD symptoms. Unfortunately, the frequency of these abiotic conditions occurring is likely to increase according to the recent consensus scenario of climate change, especially in wine-growing areas. In this article, following a review on the impact of combined thermal and water stresses on grapevine physiology, we will examine (1) how this combination of stresses might influence the lifestyle of GTD pathogens, (2) learnings from grapevine field experiments and modelling aiming at studying biotic and abiotic stresses, and (3) what mechanistic concepts can be used to explain how these stresses might affect the grapevine plant status.


Optimum Management of Overactive Bladder: Medication vs Botox® vs InterStim® vs Urgent® PC.
Optimal management of the overactive bladder is changing quickly, and urology practices need to stay in the vanguard of offering safe and effective therapies when anticholinergics are not effective or not tolerated. We will review approved therapies for overactive bladder prescribed after behavioral therapy and anticholinergic medications have failed. The treatment failure rate of anticholinergics is high and does not improve with the use of multiple drugs. Therefore, we propose a new treatment paradigm that will stop anticholinergic cycling. We believe that it is time to get patients off the anticholinergic cycle and move forward with effective alternative treatments to optimize overactive bladder therapy.


Prevention of Uterine Fibroids: molecular mechanisms and potential clinical application.
Uterine fibroids (UFs; leiomyoma) are the most common benign neoplastic threat to women worldwide, exacting an immense personal burden on female health and a monetary expense to the healthcare system estimated in the hundreds of billions of dollars every year globally. With no long-term non-invasive treatment option currently available to treat UFs, deeper insights regarding tumor etiology are the key for developing newer therapies. Accordingly, in this review, we discuss new mechanistic paradigm to explain UF tumor development through an exquisite model involving developmental reprogramming of myometrial stem cells due to early life endocrine disruptors exposure, inflammation, fibrosis, DNA damage, and eventually tissue stiffness. Further, we propose to utilize shear wave elastography as a potential screening tool for the early identification of women at risk for developing UFs who can benefit from several simple preventive strategies, including the consumption of natural compounds such as vitamin D and green tea as a safe fertility friendly non-hormonal modality to delay or even arrest or reverse UF progression.


Pathologic and molecular features of screening low-dose computed tomography (LDCT)-detected lung cancer: a baseline and 2-year repeat study.
Detailed studies on the pathologic and molecular features of low-dose computed tomography (LDCT)-detected carcinomas and comparison with unscreened tumors are still lacking. We evaluated the histopathologic features of 89 LDCT-detected lung cancers resected between 2004 and 2006. These tumors occurred within a cohort of 5202 volunteers undergoing annual LDCT, aged > or =50 years, and with a minimum 20 pack-year index. In adenocarcinomas, central scar diameter, invasion foci size and K-ras mutations were also assessed. The results were compared with those of 89 consecutive lung carcinomas matched for confounding factors (sex, smoking habit), selected from group of 363 consecutive clinically worked-up lung cancer, surgically resected in the same period and at the same Institution. The tumors were diagnosed in 63 males and 26 females (range 50-79 years), 55 of which diagnosed at the baseline (1.05%) and 34 (including 10 repeat cancers) operated after work-up during the second year (0.72%). LDCT-detected tumors showed high resectability rate (89%), earlier stage (63%) and prevalence of adenocarcinoma nodules (72%), most often of the mixed subtype, in comparison with unscreened tumors. A similar prevalence of K-ras mutations was found in both screened and unscreened adenocarcinomas. Repeat cancers were found in 10 screened patients, and were predominantly stage I adenocarcinomas of mixed subtype exhibiting smaller dimension but greater central scar diameter and stromal invasion size in comparison with the other second-year, slower-growing adenocarcinomas. Multiple tumor nodules were identified in 10 patients exclusively at the baseline, were mostly mixed adenocarcinomas and differed in their K-ras mutation profile. Screening-detected lung cancers shared most of the histologic features of fully malignant tumors, in addition to a similar prevalence of K-ras mutations, despite their earlier detection and less advanced clinical stage. Repeat cancers are potentially aggressive tumors. K-ras mutation analysis supports the impression that multifocal tumors at baseline are separate synchronous primaries.


A simple elution strategy for biotinylated proteins bound to streptavidin conjugated beads using excess biotin and heat.
Protein-protein interactions are the molecular basis of cell signaling. Recently, proximity based biotin identification (BioID) has emerged as an alternative approach to traditional co-immunoprecipitation. In this protocol, a mutant biotin ligase promiscuously labels proximal binding partners with biotin, and resulting biotinylated proteins are purified using streptavidin conjugated beads. This approach does not require preservation of protein complexes in vitro, making it an ideal approach to identify transient or weak protein complexes. However, due to the high affinity bond between streptavidin and biotin, elution of biotinylated proteins from streptavidin conjugated beads requires harsh denaturing conditions, which are often incompatible with downstream processing. To effectively release biotinylated proteins bound to streptavidin conjugated beads, we designed a series of experiments to determine optimal binding and elution conditions. Interestingly, the concentrations of SDS and IGEPAL-CA630 during the incubation with streptavidin conjugated beads were the key to effective elution of biotinylated proteins using excess biotin and heating. This protocol provides an alternative method to isolate biotinylated proteins from streptavidin conjugated beads that is suitable for further downstream analysis.


An AFLP-based survey of genetic diversity among accessions of sea oats (Uniola paniculata, Poaceae) from the southeastern Atlantic and Gulf coast states of the United States.
Uniola paniculata, commonly known as sea oats, is a C4 perennial grass capable of stabilizing sand dunes. It is most abundant along the Gulf of Mexico and southeastern Atlantic coastal regions of the United States. The species exhibits low seed set and low rates of germination and seedling emergence, and so extensive clonal reproduction is achieved through production of rhizomes, which may contribute to a decline in genetic diversity. To date, there has been no systematic assessment of genetic variability and population structure in naturally occurring stands in the USA. This study was conducted to assess the genetic relationship and diversity among nineteen U. paniculata accessions representing eight states: Texas, Louisiana, Mississippi, Alabama, Florida, South Carolina, North Carolina, and Virginia, using amplified fragment length polymorphism (AFLP). Twelve AFLP EcoRI + MseI primer combinations generated a wide range of polymorphisms (42-81%) with a mean of 59%. Overall, the sea oats plants exhibited a low range of genetic similarity. Florida accessions, FL-33 and FL-39, were most genetically diverse and the accessions from both Carolinas and Virginia (NC-1, NC-11, SC-15, and VA-53) harbored less genetic variability. Cluster analysis using the UPGMA approach separated U. paniculata plants into four major clusters which were also confirmed by principal coordinate analysis (PCO). Further examination of the different components of genetic variation by analysis of molecular variance (AMOVA) indicated the largest proportion of variability at the state level (47.8%) followed by the variation due to the differences among the genotypes within an accession (34.4%), and the differences among the accessions within a state (17.8%). The relationship between genetic diversity and geographic source of sea oats populations of the United States as revealed through this comprehensive study will be helpful to resource managers and commercial nurseries in identifying suitable plant materials for restoration of new areas without compromising the adaptation and genetic diversity.


Characterization of interaction property of multicomponents in Chinese Herb with protein by microdialysis combined with HPLC.
Interaction of traditional Chinese Herb Rhizoma Chuanxiong and protein was studied by microdialysis coupled with high performance liquid chromatography. Compounds in Rhizoma Chuanxiong, such as ferulic acid, senkyunolide A and 3-butylphthalide, were identified by HPLC, HPLC-MS and UV-vis. Microdialysis recoveries and binding degrees of compounds in Rhizoma Chuanxiong with human serum albumin (HSA) and other human plasma protein were determined: recoveries of microdialysis sampling ranged from 36.7 to 98.4% with R.S.D. below 3.1%; while binding to HSA ranged from 0 to 91.5% (0.3mM HSA) and from 0 to 93.5% (0.6mM HSA), respectively. Compared with HSA, most of compounds bound to human blood serum more extensively and the results showed that binding of these compounds in Rhizoma Chuanxiong was influenced by pH. Two compounds were found to bind to HSA and human blood serum, their binding degrees were consistent with ferulic acid and 3-butylphthalide, the active compounds in Rhizoma Chuangxiong.


Genotypic and phenotypic diversity of Lactobacillus delbrueckii subsp. lactis strains of dairy origin.
Eighty-nine strains of Lactobacillus delbrueckii subsp. lactis isolated from Italian hard and semi-hard cheeses and artisan starter cultures were characterised by phenotypic and genotypic methods. Phenotypic diversity was evaluated by studying biochemical characteristics (i.e. acidifying and peptidase activities) of technological interest. Genotypic diversity was evidenced by RAPD-PCR and pulsed field gel electrophoresis (PFGE). Phenotypic characterisation indicated a wide variability of the acidifying activity within Lact. delbrueckii subsp. lactis. Although the data was variable, it allowed us to evidence groups of strains with different acidifying properties, especially in terms of acidification intensity. Concerning peptidase activity, Lact. delbrueckii subsp. lactis showed a homogeneously high x-prolil-dipeptidil-aminopeptidase activity and a considerable but more heterogeneous lysil-aminopeptidase activity. The increased resolution obtained by the use of two molecular typing techniques, i.e. RAPD-PCR and PFGE, allowed to widen the level of strain heterogeneity. Technological and ecological pressures are determinant in selecting Lact. delbrueckii subsp. lactis sub-populations which are more functional to the different cheese technologies.


A spatiotemporally resolved infection risk model for airborne transmission of COVID-19 variants in indoor spaces.
The classic Wells-Riley model is widely used for estimation of the transmission risk of airborne pathogens in indoor spaces. However, the predictive capability of this zero-dimensional model is limited as it does not resolve the highly heterogeneous spatiotemporal distribution of airborne pathogens, and the infection risk is poorly quantified for many pathogens. In this study we address these shortcomings by developing a novel spatiotemporally resolved Wells-Riley model for prediction of the transmission risk of different COVID-19 variants in indoor environments. This modelling framework properly accounts for airborne infection risk by incorporating the latest clinical data regarding viral shedding by COVID-19 patients and SARS-CoV-2 infecting human cells. The spatiotemporal distribution of airborne pathogens is determined via computational fluid dynamics (CFD) simulations of airflow and aerosol transport, leading to an integrated model of infection risk associated with the exposure to SARS-CoV-2, which can produce quantitative 3D infection risk map for a specific SARS-CoV-2 variant in a given indoor space. Application of this model to airborne COVID-19 transmission within a hospital ward demonstrates the impact of different virus variants and respiratory PPE upon transmission risk. With the emergence of highly contagious SARS-CoV-2 variants such as the Delta and Omicron strains, respiratory PPE alone may not provide effective protection. These findings suggest a combination of optimal ventilation and respiratory PPE must be developed to effectively control the transmission of COVID-19 in healthcare settings and indoor spaces in general. This generalised risk estimation framework has the flexibility to incorporate further clinical data as such becomes available, and can be readily applied to consider a wide range of factors that impact transmission risk, including location and movement of infectious persons, virus variant and stage of infection, level of PPE and vaccination of infectious and susceptible individuals, impacts of coughing, sneezing, talking and breathing, and natural and mechanised ventilation and filtration.