Trends in Health Behaviour in Czech School-aged Children: HBSC Study. Preface.
The Health Behaviour in School-aged Children (HBSC) study is now in its 34th year. It continues to grow not only as a source of reliable and valid data on the health behaviours of 11-, 13- and 15-year-olds, but also regarding its membership - 46 countries and regions across Europe and North America are now included. The HBSC study is a cohort cross-sectional questionnaire-based study focusing on health and health-related behaviours in adolescents in the context of their social environment. This age was selected because of the significant cognitive, psychological, and biological processes taking place in this period of the lifespan, as well as because of the fact that health-related behaviours adopted at this age are known to persist into adulthood. The findings from the HBSC study are used to monitor health, well-being, and health-related behaviours in school-aged children; deepen the understanding of the social determinants of health; and provide evidence-based data for policy makers and practitioners aiming to improve the lives of young people.


Selective removal of composite sealants with near-ultraviolet laser pulses of nanosecond duration.
It is often necessary to replace pit and fissure sealants and composite restorations. This task is complicated by the necessity for complete removal of the remaining composite to enable suitable adhesion of new composite. Previous studies have shown that 355-nm laser pulses from a frequency-tripled Nd:YAG laser can selectively remove residual composite after orthodontic bracket removal on enamel surfaces. Our objective is to determine if such laser pulses are suitable for selective removal of composite pit and fissure sealants and restorations. Optical coherence tomography is used to acquire optical cross sections of the occlusal topography nondestructively before sealant application, after sealant application, and after sealant removal. Thermocouples are used to monitor the temperature in the pulp chamber during composite removal under clinically relevant ablation rates, i.e., 30 Hz and 30 mJ/pulse. At an irradiation intensity of 1.3 J/cm2, pit and fissure sealants are completely removed without visible damage to the underlying enamel. At intensities above 1.5 J/cm2, incident laser pulses remove the resin layer while at the same time preferentially etching the surface of the enamel. Temperature excursions in the pulp chamber of extracted teeth are limited to less than 5 degrees C if air-cooling is used during the rapid removal (1 to 2 min) of sealants, water-cooling is not necessary. Selective removal of composite restorative materials is possible without damage to the underlying sound tooth structure.


Report of eight children with amitraz intoxication.
Many pesticides are formulated in organic solvents. An example is amitraz, one of the formamidine groups of pesticidal chemicals. It is commonly used for the treatment of generalized demodicosis in dogs and for the control of ticks and mites in cattle and sheep. In this article, the clinical and laboratory findings of eight children with amitraz intoxication are reviewed. The purpose was to enlighten the findings of amitraz intoxication in children. Of the eight patients, five (62.5%) were boys, three (37.5%) were girls, and the ages ranged from 1 to 4 years. All children accidentally ingested amitraz orally, with no dermal exposure. The most common observed signs were decreased consciousness and bradycardia. Leukocytosis, hyperglycemia, hypernatremia, increased serum aspartate transaminase level, and prolonged partial prothrombin time were diagnosed in children. None of the children had hypothermia, hypotension, or convulsion and none of the patients died. The findings show that the initial signs and symptoms of acute amitraz intoxication appeared severe but they disappeared, with only supportive care needed in most cases within a few days.


Semimajor axis mobility of asteroidal fragments
The semimajor axes of asteroids up to about 20 kilometers in diameter drift as a result of the Yarkovsky effect, a subtle nongravitational mechanism related to radiation pressure recoil on spinning objects that orbit the sun. Over the collisional lifetimes of these objects (typically, 10 to 1000 million years), orbital semimajor axes can be moved by a few hundredths of an astronomical unit for bodies between 1 and 10 kilometers in mean radius. This has implications for the delivery of multikilometer near-Earth asteroids, because the Yarkovsky drift drives many small main-belt asteroids into the resonances that transport them to the Mars-crossing state and eventually to near-Earth space. Recent work has shown that, without such a drift, the Mars-crossing population would be depleted over about 100 million years, a time scale much smaller than the age of the solar system. Moreover, the Yarkovsky semimajor axis mobility may spread in an observable way the tight semimajor axis clustering of small asteroids produced as a consequence of disruptive collisions.


Social isolation during the COVID-19 pandemic is associated with the decline in cognitive functioning in young adults.
Coronaviruses have caused widespread disease and death worldwide, leading to the implementation of lockdown measures and the closure of educational institutions in various countries. This research aims to investigate the impact of social isolation on the cognitive functioning of young students. The study included 84 subjects, with 48 being socially isolated and 36 non-isolated individuals. The participants' mental health was assessed using the Mini-Mental State Examination (MMSE), while cognitive functions were evaluated through attention-switching tasks (AST), pattern recognition memory (PRM), and choice reaction time (CRT) tests utilizing the Cambridge Neuropsychological Automated Battery (CANTAB) software. The socially isolated group had an average age of 21.3 ± 1.1 years, whereas the non-isolated group had an average age of 22.8 ± 2.0 years. The MMSE scores were 25.8 ± 1.6 for the socially isolated group and 28.6 ± 1.3 for the non-isolated group. In terms of cognitive functioning, there were significant differences (p = 0.000) observed in the values of AST correct latency for non-switching blocks (blocks 3 and 5) between the socially isolated group (608.1 ± 139.2) and the non-isolated group (499.5 ± 67.8). Similarly, the AST mean correct latency for switching blocks (block 7) was significantly different (p = 0.012) between the socially isolated group (784.4 ± 212.5) and the non-isolated group (671.8 ± 175.6). The socially isolated group exhibited significantly higher values in AST correct mean latency, AST congruent mean latency, AST incongruent mean latency, and AST percent mean correct trials compared to the non-isolated group. Additionally, the PRM mean percent correct significantly differed (p = 0.000) between the isolated group (81.3 ± 12.0) and the non-isolated group (91.9 ± 9.2). The isolated group also showed a higher CRT correct mean latency (482.4 ± 128.9) than the non-isolated group (451.0 ± 59.0), however the difference was not significant. In conclusion, social isolation during the COVID-19 pandemic has resulted in a decline in the cognitive functioning of young students.


Medial meniscus transplantation using cyanoacrylate in rabbits.
To evaluate meniscal transplantation using as fixation method a synthetic glue derived from cyanoacrylate acid. Twenty rabbits were used, of which 10 for autologous transplantation and 10 for homologous transplantation. For the autologous transplantation the meniscus was removed, and then transplanted in the same animal, using the synthetic glue. For the homologous transplantation, the study was divided into two stages: 1--Removal of the meniscus which was maintained at a temperature of 73 C. 2--Use of cyanoacrylate acid-derived surgical adhesive for meniscal retransplantation in a different rabbit 30 days after the transplant. Due to complications, euthanasia had to be anticipated to the 15th day in the homologous group and to the 18th day in the autologous group. Macroscopically, knees submitted to transplantation presented whitish secretions from the surgical incision up to deep planes. Necrosis was observed in both groups. Statistical analysis has shown that mild (p=0.043) and moderate (p=0.001) complications emerged in a significantly earlier way in the homologous group, where euthanasia was also performed earlier (p=0.005). Synthetic surgical adhesives derived from cyanoacrylate acid promoted cortical to medullary bone necrosis bone in both groups.


Sweet Thermal Taste: Perceptual Characteristics in Water and Dependence on TAS1R2/TAS1R3.
The initial objective of this study was to determine if activation of the sweet taste receptor TAS1R2/TAS1R3 is necessary for perception of sweet thermal taste (swTT). Our approach was to inhibit the receptor with the inverse agonist lactisole using a temperature-controlled flow gustometer. Because all prior studies of thermal taste (TT) used metal thermodes to heat the tongue tip, we first investigated whether it could be generated in heated water. Experiment 1 showed that sweetness could be evoked when deionized water was heated from 20 to 35 °C, and testing with static temperatures between 20 and 35 °C demonstrated the importance of heating from a cool temperature. As in previous studies, thermal sweetness was reported by only a subset of participants, and replicate measurements found variability in reports of sweetness across trials and between sessions. Experiment 2 then showed that exposure to 8 mM lactisole blocked perception of swTT. Confirmation of the involvement of TAS1R2/TAS1R3 led to an investigation of possible sensory and cognitive interactions between thermal and chemical sweetness. Using sucrose as a sweet stimulus and quinine as a nonsweet control, we found that dynamic heating capable of producing thermal sweetness did not increase the sweetness of sucrose compared with static heating at 35 °C. However, swTT was disrupted if trials containing sucrose (but not quinine) were interspersed among heating-only trials. These findings provide new information relevant to understanding the perceptual processes and receptor mechanisms of swTT, as well as the heat sensitivity of sweet taste in general.


Sexualized drug use among gay men and other men who have sex with men in Latin America: A description of the phenomenon based on the results of LAMIS-2018.
Sexualized drug use (SDU) to enhance and extend sexual relations may involve risks of substances abuse (intoxication, interactions and overdose) and higher exposure to HIV and other sexually transmitted infections. There are inconsistencies in the methodology and findings of previous research on SDU in Latin America (LA), and more studies are required. The purpose of this research was to characterize SDU in gay men and other men who have sex with men from 18 LA countries, and describe the aspects by comparing people who practice and do not practice SDU, at the general and country levels. Cross-sectional study based on the data collected by LAMIS-2018. Dependent variable was SDU (last 12 months), and the independent variables were: drug use (in any context/in sexual context), sociodemographic, socioepidemiological, and psychosocial aspects. A descriptive analysis was carried out, comparing those who practiced and did not practice SDU. LAMIS-2018 included 64,655 participants, averaging 30 years of age. 13.6% declared having practiced SDU (6.6% with multiple partners). In the last sexual encounter the most commonly used drugs were cannabis (9.3%), poppers (6%), and Viagra (5.4%), and in the last encounter with multiple partners, poppers (19.7%), cannabis (17%), and Viagra (13.2%). HIV diagnosis was reported by 27% of people practicing SDU, vs. 14.3% in the other group. Severe anxiety-depression symptoms were more common among people practicing SDU (9.2% vs. 7%), as were the episodes of homophobic intimidation (52.6% vs. 48.2%), insults (34.4% vs. 28.6%), and aggression (4.1% vs. 3.0%). SDU was reported by a high percentage of people, with a predominance of the use of drugs related to sexual practice, and others for recreational use. Aspects described as the higher proportion of self-reported HIV diagnosis and severe symptoms of anxiety-depression among those who practiced SDU, show that is necessary to implement preventive strategies to reduce the harmful impacts that can sometimes result from this practice, including harm reduction policies, promote access to mental health services and support in situations of homophobia and stigma.


miR‑647 and miR‑1914 promote cancer progression equivalently by downregulating nuclear factor IX in colorectal cancer.
MicroRNAs (miRNAs/miRs) have been investigated as diagnostic and prognostic biomarkers for cancer; however, the significance of miRNAs in colorectal cancer (CRC) remains to be elucidated. The aim of the present study was to determine the genetic profiles of CRC tissue, and screen for miRNAs implicated in CRC cell proliferation and migration. RNA sequencing of 10 paired specimens was performed to for screen genes that were upregulated or downregulated in CRC. miRNA expression in CRC specimens and cell lines was confirmed using qPCR analysis. The significance of indicated miRNAs in CRC cell proliferation and migration was evaluated using MTT and scratch wound‑healing assays. Online computational prediction, isobaric tags for relative and absolute quantification analysis and a luciferase reporter assay were applied to determine candidate targeted genes for the miRNAs. RNA‑seq data revealed miR‑1914 as the most prominent miRNA in CRC specimens. qPCR analysis also suggested that the expression of miR‑1914, as well as its counterpart miR‑647 were elevated in CRC specimens and cell lines. Suppression of miR‑647/1914 using small interfering RNAs inhibited CRC SW480 and SW620 cell proliferation, and migration. Nuclear factor I/X (NFIX) was demonstrated to be a candidate for miR‑647/1914 and mediated the oncogenic activity of miR‑647/1914. In all, miR‑647 and miR‑1914 were demonstrated to promote the proliferation and migration of CRC cells by directly targeting NFIX. Therapeutic delivery of siRNAs targeting miR‑647/1914 and overexpression of NFIX may be feasible approaches for CRC treatment.


A secretory protein involved in the antagonistic interactions between methanotrophic bacteria.
Antagonistic interactions in mixed culture of methanotrophic bacteria Methylomonas methanica 12 and Methylocystis minimus 33 were investigated. The inhibitory action of Mcs. minimus exometabolites against Mm. methanica grown in liquid medium was found to be specific. Ultrafiltration established that the molecular weight of the substance having inhibitory activity lies within the range 2-10 kD. The activity is protease sensitive and relatively stable to heating. Electrophoretic analysis showed that a protein with molecular weight of approximately 8 kD prevailed in Mcs. minimus culture liquid. When Mm. methanica cells were incubated in culture liquid of Mcs. minimus, the sorption of the 8 kD protein by target cells was observed. This suggests that the inhibitory effect may be associated with the 8 kD protein which has properties similar to known bacteriocins.


Feeding steam-pelleted rapeseed affects expression of genes involved in hepatic lipid metabolism and fatty acid composition of chicken meat.
This study investigated the dietary effect of steam-pelleted rapeseed (RS) diets with different inclusion levels on the fatty acid composition of chicken meat and the expression of lipid metabolism-related genes in the liver. Experimental diets included 6 different wheat-soybean meal based diets either in nonpelleted or steam-pelleted form supplemented with 80, 160, and 240 g RS/kg feed and one nonpelleted wheat-soybean meal based diet without RS supplementation as the control. These diets were fed to newly hatched broiler chickens (Ross 308) for 34 days. Compared to the control diet, steam-pelleted diets containing 160 or 240 g/kg RS significantly increased the content of omega-3 long chain polyunsaturated fatty acids (n-3 LC-PUFA) in the breast and drumstick, while their meat yields were not affected. Moreover, the mRNA levels of fatty acid desaturase 1 (FADS1) and acyl-coenzyme A oxidase 1 (ACOX1) in their livers increased. Therefore, steam-pelleted diets with 160 or 240 g/kg RS can be used to increase the n-3 LC-PUFA content in chicken meat without compromising meat yield.


Sugar consumption in 5 and 12-year-old school children in Ondo State, Nigeria in 1985.
In a previous study (Olojugba & Lennon, 1987) of 5 and 12-year-old children residing in Ondo State, Nigeria, it was shown that caries had increased (1977 to 1983) in children of both age groups in higher social class families; caries had also increased in the 12-year-old children but not in those aged 5 year olds among lower social class families. The object of the present study was to examine sugar consumption in samples of children of the same age groups drawn from the same population. One hundred families with siblings in both age groups were selected. 24-hour dietary records were obtained for each child; an inventory was also made of all food and drink stored within the home. The 5-year-old children from the higher social class families had 5.9 sugar intakes, as against 1.5 intakes among their lower social class urban counterparts. All the sugar-containing items were consumed by the 5-year-old children at home. The difference in the frequency of intake among those aged 12 years was much less obvious, 4.9 and 4.2 intakes respectively. Sixty-eight per cent of the intakes among the lower social class 12-year-old children was in the form of cheap sugar-containing drinks, ice lollies and sweets which they bought themselves and consumed away from home. These cheap sources of sugar were not available to their younger siblings. It was concluded that the sugar consumption patterns recorded in the children, and confirmed by the household food inventory, provided a plausible explanation of the previously reported changes in caries severity.


Runx1 deletion or dominant inhibition reduces Cebpa transcription via conserved promoter and distal enhancer sites to favor monopoiesis over granulopoiesis.
Deletion of Runx1 in adult mice produces a myeloproliferative phenotype. We now find that Runx1 gene deletion increases marrow monocyte while reducing granulocyte progenitors and that exogenous RUNX1 rescues granulopoiesis. Deletion of Runx1 reduces Cebpa mRNA in lineage-negative marrow cells and in granulocyte-monocyte progenitors or common myeloid progenitors. Pu.1 mRNA is also decreased, but to a lesser extent. We also transduced marrow with dominant-inhibitory RUNX1a. As with Runx1 gene deletion, RUNX1a expands lineage-Sca-1+c-kit+ and myeloid cells, increased monocyte CFUs relative to granulocyte CFUs, and reduced Cebpa mRNA. Runx1 binds a conserved site in the Cebpa promoter and binds 4 sites in a conserved 450-bp region located at +37 kb; mutation of the enhancer sites reduces activity 6-fold in 32Dcl3 myeloid cells. Endogenous Runx1 binds the promoter and putative +37 kb enhancer as assessed by ChIP, and RUNX1-ER rapidly induces Cebpa mRNA in these cells, even in cycloheximide, consistent with direct gene regulation. The +37 kb region contains strong H3K4me1 histone modification and p300-binding, as often seen with enhancers. Finally, exogenous C/EBPα increases granulocyte relative to monocyte progenitors in Runx1-deleted marrow cells. Diminished CEBPA transcription and consequent impairment of myeloid differentiation may contribute to leukemic transformation in acute myeloid leukemia cases associated with decreased RUNX1 activity.


Role of complement in inflammation and injury in the nervous system.
The complement (C) system plays important roles in host defence but activation at inappropriate sites or to an excessive degree can cause host tissue damage. C has been implicated as a factor in the causation or propagation of tissue injury in numerous diseases. The brain is an immunologically isolated site, sheltered from circulating cells and proteins of the immune system; nevertheless, there is a growing body of evidence implicating C in numerous brain diseases. In this brief article we review the evidence suggesting a role for C in diseases of the central and peripheral nervous system and discuss the possible sources of C at these sites. Some brain cells synthesise C and also express specific receptors; some are exquisitely sensitive to the lytic effects of C. The evidence suggests that C synthesis and activation in the brain are important in immune defence at this site but may also play a role in brain disease.


[External and intracavitary thermometry and the temperature sensitivity of the bladder in children].
The authors have determined specification indices of the intracavitary temperature of the urinary bladder in children of different age, which are compared with data of "reflected" temperature from the urinary bladder onto the abdominal wall which allows objective determination and control in dynamics of the course of the inflammatory process in the urinary bladder. The thermocontrast "cold" and "hyperthermic" cystotonometry are new methods of investigation allowing to objectively determine the state of the cold and warm receptors of the urinary bladder, to detect latent neurogenic disturbances of the urinary bladder and to prognose the effectiveness of treatment of hypermotor disturbances of functions of the urinary bladder.


Transferrin receptors of human fibroblasts. Analysis of receptor properties and regulation.
Normal human skin fibroblasts cultured in vitro exhibit specific binding sites for 125I-labelled transferrin. Kinetic studies revealed a rate constant for association (Kon) at 37 degrees C of 1.03 X 10(7) M-1 X min-1. The rate constant for dissociation (Koff) at 37 degrees C was 7.9 X 10(-2) X min-1. The dissociation constant (KD) was 5.1 X 10(-9) M as determined by Scatchard analysis of binding and analysis of rate constants. Fibroblasts were capable of binding 3.9 X 10(5) molecules of transferrin per cell. Binding of 125I-labelled diferric transferrin to cells was inhibited equally by either apo-transferrin or diferric transferrin, but no inhibition was evident with apo-lactoferrin, iron-saturated lactoferrin, or albumin. Preincubation of cells with saturating levels of diferric transferrin or apo-transferrin produced no significant change in receptor number or affinity. Preincubation of cells with ferric ammonium citrate caused a time- and dose-dependent decrease in transferrin binding. After preincubation with ferric ammonium citrate for 72 h, diferric transferrin binding was 37.7% of control, but no change in receptor affinity was apparent by Scatchard analysis. These results suggest that fibroblast transferrin receptor number is modulated by intracellular iron content and not by ligand-receptor binding.


Data verification at health centers and district health offices in Xiengkhouang and Houaphanh Provinces, Lao PDR.
Routine health information is an essential health system building block. In low and low-middle income countries however, concerns about the quality of routine administrative data have often undermined their use. The purpose of the present study was to verify the data availability, and consistency of six key maternal and child health indicators (first antenatal care, fourth antenatal care, skilled birth attendants, postnatal care, 'Bacillus Calmette Guerin and diphtheria-pertussis-tetanus third dose). The study collected data for the identified indicators in 2011 from Xiengkhouang and Houaphanh provinces in the Lao People's Democratic Republic (PDR). The data came from health centres (N = 109), sub-districts (N = 26) and district health offices (N = 16). Core indicators were calculated using numerators and denominators from the different data sources at the district and health centre level and standardized statistical tests performed. The study revealed that data for the six indicators were either not available or not complete in the service logbooks or registers in most of the health centres. Furthermore, few health centres kept the data for up to five years, often destroying it once the report had been sent to the district health office. In addition, there was limited numerator consistency between the different data sources. Data on the six indicators collected and reported in the public health system across the two provinces lacked completeness, accuracy and consistency. To improve the quality of data, there is a need to train health centre staff in data collection and recording as well as ensuring there is adequate monitoring and supervision. A uniform national standardized form is also necessary with findings shared with district health offices and centres. Additionally, staff should be encouraged to own and value local data.


Rhythm makes the world go round: An MEG-TMS study on the role of right TPJ theta oscillations in embodied perspective taking.
While some aspects of social processing are shared between humans and other species, some aspects are not. The former seems to apply to merely tracking another's visual perspective in the world (i.e., what a conspecific can or cannot perceive), while the latter applies to perspective taking in form of mentally "embodying" another's viewpoint. Our previous behavioural research had indicated that only perspective taking, but not tracking, relies on simulating a body schema rotation into another's viewpoint. In the current study we employed Magnetoencephalography (MEG) and revealed that this mechanism of mental body schema rotation is primarily linked to theta oscillations in a wider brain network of body-schema, somatosensory and motor-related areas, with the right posterior temporo-parietal junction (pTPJ) at its core. The latter was reflected by a convergence of theta oscillatory power in right pTPJ obtained by overlapping the separately localised effects of rotation demands (angular disparity effect), cognitive embodiment (posture congruence effect), and basic body schema involvement (posture relevance effect) during perspective taking in contrast to perspective tracking. In a subsequent experiment we interfered with right pTPJ processing using dual pulse Transcranial Magnetic Stimulation (dpTMS) and observed a significant reduction of embodied processing. We conclude that right TPJ is the crucial network hub for transforming the embodied self into another's viewpoint, body and/or mind, thus, substantiating how conflicting representations between self and other may be resolved and potentially highlighting the embodied origins of high-level social cognition in general.


[Is it necessary to discontinue captopril before heart surgery?].
Interaction between kininase II and anaesthesia is not well described. Twenty two patients treated by kininase II for congestive heart failure are studied during anaesthesia for cardiovascular surgery. A first group of seventeen homogeneous hemodynamic data are reported. High cardiac index contrasts with severe clinical cardiac failure. A second group of inhomogeneous patients are separately described. Vasoconstrictor can be codified in the situation of low systemic resistance with high cardiac index. Preoperative treatment can be continued, under requirement of hemodynamic monitoring.


A spiral tack as a lead point for volvulus.
We present a case of small bowel volvulus around an endotack applied during total extraperitoneal laparoscopic hernia repair (TEP). This complication prompts reconsideration of the role of tacks during extraperitoneal laparoscopic hernia repairs. We undertook a chart review and provide a case presentation with review of the literature. A 66-year-old male with bilateral inguinal hernias underwent elective, bilateral, total extraperitoneal laparoscopic hernia repair. During dissection, a small peritoneal tear occurred. The tear was closed with a spiral tack. On postoperative day 22, the patient developed an acute abdomen. Exploratory laparotomy revealed a volvulus rotated around an adhesion to the spiral tack. Volvulus can cause vascular compromise leading to bowel ischemia and necrosis. A tack resulting in adhesion and volvulus is an unusual, but serious, complication. Repair of a peritoneal tear during preperitoneal hernia repair is advocated to improve visualization obstructed by a pneumoperitonuem and decrease adhesions to the abdominal wall. The use of blunt Endoloops or crimps may prove safer than tacks for repairing the peritoneum and placement in proximity to delicate or thin tissues. Additionally, careful placement of foreign bodies to ensure their stability and to minimize protrusion may decrease the risk of erosion of the hardware.


Calculation of Chemical Reaction Barrier Heights by Multiconfiguration Pair-Density Functional Theory with Correlated Participating Orbitals.
The accurate description of reaction barrier heights is challenging for quantum mechanical methods due to the need for a balanced treatment of dynamic and static correlation energies because their importance varies during the course of a chemical reaction. While some regions of potential energy surfaces are well-described by a single-reference wave function or by Kohn-Sham density functional theory, in other cases a multireference treatment is needed. For systems with many active electrons, most accurate multireference methods have prohibitive computational scalings with system size. Multiconfiguration pair-density functional theory, MC-PDFT, is a more affordable multireference approach that computes the total electron correlation energy in a single step by using the multiconfiguration kinetic energy, density, and on-top pair density and an on-top density functional. In this work, we apply MC-PDFT to a benchmark database (DBH24/18) of 24 diverse reaction barrier heights. We explore the role of active space and basis set selection on the performance of MC-PDFT. We find that MC-PDFT is able to calculate reaction barrier heights with a similar accuracy to complete active space second order perturbation theory, CASPT2, but at a lower computational cost, and we find that MC-PDFT is less dependent on basis set selection than CASPT2.


Modeling Operando Electrochemical CO2 Reduction.
Since the seminal works on the application of density functional theory and the computational hydrogen electrode to electrochemical CO2 reduction (eCO2R) and hydrogen evolution (HER), the modeling of both reactions has quickly evolved for the last two decades. Formulation of thermodynamic and kinetic linear scaling relationships for key intermediates on crystalline materials have led to the definition of activity volcano plots, overpotential diagrams, and full exploitation of these theoretical outcomes at laboratory scale. However, recent studies hint at the role of morphological changes and short-lived intermediates in ruling the catalytic performance under operating conditions, further raising the bar for the modeling of electrocatalytic systems. Here, we highlight some novel methodological approaches employed to address eCO2R and HER reactions. Moving from the atomic scale to the bulk electrolyte, we first show how ab initio and machine learning methodologies can partially reproduce surface reconstruction under operation, thus identifying active sites and reaction mechanisms if coupled with microkinetic modeling. Later, we introduce the potential of density functional theory and machine learning to interpret data from Operando spectroelectrochemical techniques, such as Raman spectroscopy and extended X-ray absorption fine structure characterization. Next, we review the role of electrolyte and mass transport effects. Finally, we suggest further challenges for computational modeling in the near future as well as our perspective on the directions to follow.


Effectiveness of proprotein convertase subtilisin/kexin-9 monoclonal antibody treatment on plasma lipoprotein(a) concentrations in patients with elevated lipoprotein(a) attending a clinic.
Lipoprotein(a) (Lp[a]) is a causal risk factor for atherosclerotic cardiovascular disease (ASCVD). Proprotein convertase subtilisin/kexin-9 monoclonal antibodies (PCSK9mAbs) can lower Lp(a) levels in clinical trials, but their effects in patients with elevated Lp(a) in clinical practice remain unclear. To investigate the effectiveness and safety of PCSK9mAbs in lowering plasma Lp(a) in patients with elevated Lp(a) concentrations in a lipid clinic. This was an open-label study of 53 adult patients with elevated Lp(a) concentration (≥0.5 g/L). Clinical, biochemical, and safety data were collected before and on treatment with evolocumab or alirocumab over a mean period of 11 months. Treatment with a PCSK9mAb resulted in a significant reduction of 0.29 g/L (-22%) in plasma Lp(a) concentration (p<.001). There were also significant reductions in low-density lipoprotein-cholesterol (LDL-C) (-53%), remnant-cholesterol (-12%) and apolipoprotein B (-43%) concentrations. The change in Lp(a) concentration was significantly different from a comparable group of 35 patients with elevated Lp(a) who were not treated with a PCSK9mAb (-22% vs. -2%, p<.001). The reduction in Lp(a) concentration was not associated with the corresponding changes in LDL-C, remnant-cholesterol, and apolipoprotein B (p>.05 in all). 7.5% and 47% of the patients attained a target concentration of Lp(a) <0.5 g/L and LDL-C <1.8 mmol/L, respectively. PCSK9mAbs were well tolerated, the common adverse effects being pharyngitis (9.4%), nasal congestion (7.6%), myalgia (9.4%), diarrhoea (7.6%), arthralgia (9.4%) and injection site reactions (11%). PCSK9mAbs can effectively and safely lower plasma Lp(a) concentrations in patients with elevated Lp(a) in clinical practice; the impact of the fall in Lp(a) on ASCVD outcomes requires further investigation.


Oedema on STIR modified the effect of amoxicillin as treatment for chronic low back pain with Modic changes-subgroup analysis of a randomized trial.
To evaluate potential MRI-defined effect modifiers of amoxicillin treatment in patients with chronic low back pain and type 1 or 2 Modic changes (MCs) at the level of a previous lumbar disc herniation (index level). In a prospective trial (AIM), 180 patients (25-64 years; mean age 45; 105 women) were randomised to receive amoxicillin or placebo for 3 months. Primary outcome was the Roland-Morris Disability Questionnaire (RMDQ) score (0-24 scale) at 1 year. Mean RMDQ score difference between the groups at 1 year defined the treatment effect; 4 RMDQ points defined the minimal clinically important effect. Predefined baseline MRI features of MCs at the index level(s) were investigated as potential effect modifiers. The predefined primary hypothesis was a better effect of amoxicillin when short tau inversion recovery (STIR) shows more MC-related high signal. To evaluate this hypothesis, we pre-constructed a composite variable with three categories (STIR1/2/3). STIR3 implied MC-related STIR signal increases with volume ≥ 25% and height > 50% of vertebral body and maximum intensity increase ≥ 25% and presence on both sides of the disc. As pre-planned, interaction with treatment was analysed using ANCOVA in the per protocol population (n = 155). The STIR3 composite group (n = 41) and STIR signal volume ≥ 25% alone (n = 45) modified the treatment effect of amoxicillin. As hypothesised, STIR3 patients reported the largest effect (- 5.1 RMDQ points; 95% CI - 8.2 to - 1.9; p for interaction = 0.008). Predefined subgroups with abundant MC-related index-level oedema on STIR modified the effect of amoxicillin. This finding needs replication and further support. • In the primary analysis of the AIM trial, the effect of amoxicillin in patients with chronic low back pain and type 1 or 2 MCs did not reach the predefined cut-off for clinical importance. • In the present MRI subgroup analysis of AIM, predefined subgroups with abundant MC-related oedema on STIR reported an effect of amoxicillin. • This finding requires replication and further support.


Scattering-Based Light-Sheet Microscopy for Rapid Cellular Imaging of Fresh Tissue.
Light-sheet microscopy (LSM) is a novel imaging technology that has been used for imaging fluorescence contrast in basic life science research. In this paper, we have developed a scattering-based LSM (sLSM) for rapidly imaging the cellular morphology of fresh tissues without any exogenous fluorescent dyes. In the sLSM device, a thin light sheet with the central wavelength of 834 nm was incident on the tissue obliquely, 45° relative to the tissue surface. The detection optics was configured to map the light sheet-illuminated area onto a two-dimensional imaging sensor. The illumination numerical aperture (NA) was set as 0.0625, and the detection NA 0.3. The sLSM device achieved a light sheet thickness of less than 6.7 µm over 284 µm along the illumination optical axis. The detection optics of the sLSM device had a resolution of 1.8 µm. The sLSM images of the swine kidney ex vivo visualized tubules with similar sizes and shapes to those observed in histopathologic images. The swine duodenum sLSM images revealed cell nuclei and villi architecture in superficial lesions and glands in deeper regions. The preliminary results suggest that sLSM may have the potential for rapidly examining the freshly-excised tissue ex vivo or intact tissue in vivo at microscopic resolution. Further optimization and performance evaluation of the sLSM technology will be needed in the future. Lasers Surg. Med. © 2020 Wiley Periodicals LLC.


Controlling beta-sheet assembly in genetically engineered silk by enzymatic phosphorylation/dephosphorylation.
Enzymatic phosphorylation and dephosphorylation reactions were used to modify a genetically engineered variant of spider dragline silk. The approximately 25 kDa protein was phosphorylated with cyclic AMP-dependent kinase and dephosphorylated with calf intestinal alkaline phosphatase. Phosphorylation inhibited beta-sheet assembly of the protein and enhanced solubility to about 5 mg/mL in water, compared to about 20% of this level upon enzymatic dephosphorylation. The cyclability of the phosphorylation-dephosphorylation system was confirmed by MALDI with a model peptide. Kinetic studies conducted with [gamma-(32)P]ATP illustrate that the phosphorylation reaction proceeds over 6 h. Secondary structure of the phosphorylated and dephosphorylated proteins was determined by CD and FTIR. The results illustrate that an enzymatic phosphorylation event can be used to control the solution structure of a protein like silk, which has a tendency to prematurely precipitate due to the formation of beta-sheets.


The multiscale coarse-graining method. I. A rigorous bridge between atomistic and coarse-grained models.
Coarse-grained (CG) models provide a computationally efficient method for rapidly investigating the long time- and length-scale processes that play a critical role in many important biological and soft matter processes. Recently, Izvekov and Voth introduced a new multiscale coarse-graining (MS-CG) method [J. Phys. Chem. B 109, 2469 (2005); J. Chem. Phys. 123, 134105 (2005)] for determining the effective interactions between CG sites using information from simulations of atomically detailed models. The present work develops a formal statistical mechanical framework for the MS-CG method and demonstrates that the variational principle underlying the method may, in principle, be employed to determine the many-body potential of mean force (PMF) that governs the equilibrium distribution of positions of the CG sites for the MS-CG models. A CG model that employs such a PMF as a "potential energy function" will generate an equilibrium probability distribution of CG sites that is consistent with the atomically detailed model from which the PMF is derived. Consequently, the MS-CG method provides a formal multiscale bridge rigorously connecting the equilibrium ensembles generated with atomistic and CG models. The variational principle also suggests a class of practical algorithms for calculating approximations to this many-body PMF that are optimal. These algorithms use computer simulation data from the atomically detailed model. Finally, important generalizations of the MS-CG method are introduced for treating systems with rigid intramolecular constraints and for developing CG models whose equilibrium momentum distribution is consistent with that of an atomically detailed model.


Vinculin and talin: focus on the myocardium.
Cardiomyopathy is a heart muscle disease caused by decreased contractility of the ventricles leading to heart failure and premature death. Multiple conditions like ischemic heart disease (atherosclerosis), hypertension, diabetes, viral infection, alcohol abuse, obesity and genetic mutations can lead to cardiomyopathy. Single gene mutations in sarcomeric proteins, Z-disk-associated proteins, membrane/associated proteins, intermediate filaments, calcium cycle proteins as well as in modifier genes have been linked to cardiomyopathy. Clinical practice guidelines have been formulated by the American Heart Association and the Heart Failure Association of America on how to genetically evaluate patients with cardiomyopathy. To illustrate the concept that alterations in genes cause cardiovascular disease, this review will focus on two membrane-associated proteins, vinculin and talin. We will discuss the general function of vinculin/metavinulin as well as talin1 and talin2, with emphasis on what is understood about their role in the cardiac myocyte and in whole heart.


Spectral narrowing of x-ray pulses for precision spectroscopy with nuclear resonances.
Spectroscopy of nuclear resonances offers a wide range of applications due to the remarkable energy resolution afforded by their narrow linewidths. However, progress toward higher resolution is inhibited at modern x-ray sources because they deliver only a tiny fraction of the photons on resonance, with the remainder contributing to an off-resonant background. We devised an experimental setup that uses the fast mechanical motion of a resonant target to manipulate the spectrum of a given x-ray pulse and to redistribute off-resonant spectral intensity onto the resonance. As a consequence, the resonant pulse brilliance is increased while the off-resonant background is reduced. Because our method is compatible with existing and upcoming pulsed x-ray sources, we anticipate that this approach will find applications that require ultranarrow x-ray resonances.


Electrostatic Interactions between Acid-/Base-Containing Polymer Nanoparticles and Proteins: Impact of Polymerization pH.
Electrostatic interaction between synthetic polymer nanoparticles (NPs) and proteins is of considerable importance in the design of NPs that capture, neutralize, and deliver target molecules in a biological milieu. Ionizable functional groups, such as carboxylic acids and amines, are often introduced to NPs to tune the affinity with target bio-macromolecules through electrostatic attraction and repulsion. However, acids/bases are not always ionized at a physiological pH because acidities of the functional groups depend on the microenvironment around the acids/bases that are imprinted during the polymerization process. Here, we show that electrostatic interaction between acid-/base-containing NPs and target proteins strongly depends on the pH of the solution during the NP polymerization process. To prepare NPs that capture target proteins by electrostatic interactions at physiological pH, NPs must be polymerized within a pH range where the acid/base monomer is ionized. Acid-/base-containing NPs that exhibit completely different interactions with the proteins can be prepared by changing the polymerization pH without changing monomer compositions. Our results indicate that polymerization pH should be carefully tuned to design acid-/base-containing NPs that show desired affinity to all proteins in a biological milieu and to proteins of interest.


[Preemptive analgesic efficacy of gabapentin and nimesulide in the functional endoscopic sinus surgery].
Preemptive analgesia is an important factor in controlling the postoperative pain and avoiding the stress response caused by the surgery. We aim to compare impact of gabapentin and nimesulide on postoperative analgesic consumption on the visual analog scale (VAS) as well as any potential side effects, to those of the placebo group. Ninety patients aged 18-70 under ASA I-II risk, who were to undergo Functional Endoscopic Sinus Surgery (FESS), were included in the study. Patients were randomized into three groups. One hour before the surgery, oral placebo capsules were administered to Group I patients, Gabapentin 1200 mg to Group II patients and Nimesulide 100 mg to Group III patients. Perioperative hemodynamic parameters were measured for all patients. During the first 24 hours of the postoperative period, side effects such as nausea, vomiting, hypotension, hypertension, bradycardia, tachycardia were noted, and the time from end of the surgery to the first diclofenac need was recorded in addition to VAS scores. In all of the follow-ups for 24 hours of the postoperative period, time from end of the surgery until the first diclofenac requirement; Group 2 > Group 3 > Group 1. Total diclofenac consumption of group 2 was found to be significantly lower than Group 1 and Group 3 on a statistical basis. Postoperative VAS score in Group 2 was lower than Group 1 and Group 3. To conclude, we believe that gabapentin or nimesulide may be safely used preemptively for the purposes of postoperative analgesia after FESS procedures.


Apparent ileal and total-tract nutrient digestion by pigs as affected by dietary nondigestible oligosaccharides.
The effects of two types of nondigestible oligosaccharides (NDO), fructooligosaccharides (FOS), and transgalactooligosaccharides (TOS) were studied on growing and weanling pigs' nutrient digestion. Dietary NDO were included at the expense of purified cellulose. Twenty-five 57-d-old growing pigs, averaging 15.9+/-.6 kg on d 0 of the experiment, were fed a corn-based control diet or the control with 6.8 or 13.5 g of FOS/kg or 4.0 or 8.0 g of TOS/kg (five pigs per diet). Feces were collected on d 28 to 32, and small-intestinal digesta were collected (slaughter technique) on d 42 to 47 of the experiment. Feeds, feces, and digesta were analyzed for DM, inorganic matter, CP, ether extract, and crude fiber. Dietary NDO did not significantly affect apparent fecal and small intestinal digestion of nutrients in growing pigs. After being fed a NDO-free diet through d 10 after weaning, 38-d-old weanling pigs (n = 20), averaging 10.4+/-.8 kg on d 0 of the experiment, were fed a control diet (based on cornstarch, casein, and oat husk meal) or the control with 10 or 40 g of FOS or TOS/kg (four pigs per diet). Feces and urine were collected on d 13 to 17, and ileal digesta were collected via a postvalve T-cecum cannula on d 33 to 37 of the experiment. Feeds, feces, and digesta were analyzed for DM, inorganic matter, CP, ether extract, starch, NDF, ADF, ADL, Ca, P, Mg, Fe, Cu, and Zn. Nonstarch neutral-detergent soluble carbohydrates (NNSC) completed the mass balance for the carbohydrates. Urine was analyzed for N and minerals. The apparent fecal digestion of NNSC increased in the NDO-supplemented diets. The TOS-fed pigs tended (P<.10) to have a higher apparent fecal digestion of CP than the FOS-fed and control pigs but excreted more N via the urine (P<.01). Nitrogen and mineral balances were not affected. The FOS was nearly completely degraded prececally. Mean fiber digestion was lower at the fecal compared with the ileal level, as was the extent of NDO effects. This indicates that fiber digestion requires more than 2 wk to adapt to dietary NDO. Apparent ileal digestion of hemicellulose increased for the NDO-supplemented diets (P<.05), but that of NNSC decreased (P<.001). Thus, under the well-controlled conditions of this experiment, dietary NDO hardly affected nutrient digestion in well-kept growing and weanling pigs. However, digestion of dietary nonstarch carbohydrates may be affected.


Asthma severity as a contributing factor to cancer incidence: A cohort study.
A putative link between asthma and asthma severity with the occurrence of cancer has been suggested but has not been fully investigated. The objective of this study is to assess the incidence of all types of cancer in a cohort of asthmatic patients. A single center cohort retrospective study was conducted to investigate the role of asthma as a potential risk factor for various cancers. Participants were followed for a period of 9 years from 01/01/2010 to 30/12/2018 and cancer incidence and its determinants were collected in asthmatic patients and controls from the same population source but without any respiratory disease. Overall, 2,027 asthma patients and 1,637 controls were followed up for an average of 9 years. The statistical analysis showed that 2% of asthma patients were diagnosed with various cancers, resulting in an incidence rate of cancer of 383.02 per 100,000 persons per year which is significantly higher than the 139.01 per 100,000 persons per year observed in matched controls (p-value < 0.001). The top four cancers reported among asthmatics were breast, colon, lung and prostate cancer. Lung cancer in asthmatics had the longest diagnosis period with a mean of 36.6 years compared to the shortest with prostate cancer with 16.5 years. This study shows that asthma patients are at increased risk of different types of cancers with asthma severity and goiter as the main factors that may increase the risk of developing cancers among asthmatic patients.


Recovery, growth, and development of Acanthoparyphium tyosenense (Digenea: Echinostomatidae) in experimental chicks.
Chicks were experimentally infected with Acanthoparyphium tyosenense (Digenea: Echinostomatidae) metacercariae per os, and the growth and development of worms in this host were observed from days I to 38 postinfection (PI). The worms grew rapidly and matured sexually in the small intestine (chiefly in the jejunum) of chicks by day 5 PI. and survived at least up to day 38 Pi, although worm recovery decreased after day 5 PI. Both parenchymal and reproductive organs increased greatly in size from day 2 to day 10 PI and then continued to increase gradually in size up to day 38 PI. The number of uterine eggs reached a peak on days 10 and 15 PI and then decreased gradually. The results suggest that chicks are a fairly suitable definitive host for experimental infection with A. tyosenense.


Structures of plant viruses from vibrational circular dichroism.
Vibrational circular dichroism (VCD) spectra in the amide I and II regions have been measured for viruses for the first time. VCD spectra were recorded for films prepared from aqueous buffer solutions and also for solutions using D(2)O buffers at pH 8. Investigations of four filamentous plant viruses, Tobacco mosaic virus (TMV), Papaya mosaic virus, Narcissus mosaic virus (NMV) and Potato virus X (PVX), as well as a deletion mutant of PVX, are described in this paper. The film VCD spectra of the viruses clearly revealed helical structures in the virus coat proteins; the nucleic acid bases present in the single-stranded RNA could also be characterized. In contrast, the solution VCD spectra showed the characteristic VCD bands for alpha-helical structures in the coat proteins but not for RNA. Both sets of results clearly indicated that the coat protein conformations are dominated by helical structures, in agreement with earlier reports. VCD results also indicated that the coat protein structures in PVX and NMV are similar to each other and somewhat different from that of TMV. The present study demonstrates the feasibility of measuring VCD spectra for viruses and extracting structural information from these spectra.


Combination effect of voluntary exercise and garlic (Allium sativum) on oxidative stress, cholesterol level and histopathology of heart tissue in type 1 diabetic rats.
Introduction: This study aimed to evaluate the combination effect of voluntary exercise and garlic on oxidative markers, cholesterol level and histological alterations in streptozotocin (STZ)- induced diabetes in rat heart. Methods: Thirty-five male Wistar rats were randomly assigned into five experimental groups (n=7): Control, Diabetes, Diabetes+Garlic, Diabetes+Exercise, Diabetes+Garlic+Exercise groups. Diabetes was induced by STZ (ip, 50 mg/kg) in animals. Animals received garlic homogenate (250 mg/kg) by oral gavage or subjected to voluntary exercise alone or together for 6 weeks. At the end of intervention blood and heart tissue samples were obtained and used for measurement of glycosylated haemoglobin (HbA1c), cholesterol, total antioxidant capacity (TAC), malondialdehyde (MDA) levels and histological analysis. Results: Improved blood glucose, cholesterol, total antioxidant capacity, and MDA levels were established in both Diabetes+Garlic and Diabetes+Exercise groups. Additionally, staining with Hematoxylin and Eosin (H&E) revealed that voluntary exercise and garlic alone and together reduced interstitial edema, leukocyte infiltration, and myonecrosis. Furthermore, simultaneous treatment of diabetic rats with garlic and exercise training had an additive effect on these parameters. Conclusion: The findings indicated that combination therapy with garlic and voluntary exercise may present more beneficial effects in heart histological remodeling in diabetic rats than the use of garlic or voluntary exercise alone and that these beneficial effects might be associated with enhancement in cholesterol, total antioxidant capacity, and MDA levels.


Sequencing of Plant Wall Heteroxylans Using Enzymic, Chemical (Methylation) and Physical (Mass Spectrometry, Nuclear Magnetic Resonance) Techniques.
This protocol describes the specific techniques used for the characterization of reducing end (RE) and internal region glycosyl sequence(s) of heteroxylans. De-starched wheat endosperm cell walls were isolated as an alcohol-insoluble residue (AIR)(1) and sequentially extracted with water (W-sol Fr) and 1 M KOH containing 1% NaBH4 (KOH-sol Fr) as described by Ratnayake et al. (2014)(2). Two different approaches (see summary in Figure 1) are adopted. In the first, intact W-sol AXs are treated with 2AB to tag the original RE backbone chain sugar residue and then treated with an endoxylanase to generate a mixture of 2AB-labelled RE and internal region reducing oligosaccharides, respectively. In a second approach, the KOH-sol Fr is hydrolyzed with endoxylanase to first generate a mixture of oligosaccharides which are subsequently labelled with 2AB. The enzymically released ((un)tagged) oligosaccharides from both W- and KOH-sol Frs are then methylated and the detailed structural analysis of both the native and methylated oligosaccharides is performed using a combination of MALDI-TOF-MS, RP-HPLC-ESI-QTOF-MS and ESI-MS(n). Endoxylanase digested KOH-sol AXs are also characterized by nuclear magnetic resonance (NMR) that also provides information on the anomeric configuration. These techniques can be applied to other classes of polysaccharides using the appropriate endo-hydrolases.


Transport of copper oxychloride-based fungicide particles in saturated quartz sand.
Intensive use of copper-based fungicides in agriculture causes contamination of subsurface environment. While the transport of dissolved copper in porous media has been widely studied, transport mechanisms of particles of copper-based fungicides are poorly understood. This paper reports the results of tests involving the transport of colloid-size particles of a copper oxychloride-based fungicide (COF) in water-saturated quartz sand columns under varying electrochemical and hydrodynamic conditions. The effect of the ionic strength on colloid attachment and exclusion suggests that interactions of COF in water depend on the characteristics of the diffuse layer. Hydrodynamic shear influences the deposition of fungicide particles, indicating that attachment forces are weak. Particle deposition dynamics was well-fitted with the two-site kinetic attachment model, which reflects the heterogeneity of the quartz surface. Retention also occurs for unfavorable electrochemical conditions, which was attributed to stagnation zones arising from the physical and chemical heterogeneity of the quartz surface.


Developing a Theoretical Framework for a Complex Maternal-Child Health Intervention: Health Beginning Initiative.
A theoretical framework serves as a roadmap for the implementation and application of a complex, health promotion intervention; is used to test hypotheses; and guides analysis and evaluation of the intervention. The purpose of this article is to demonstrate how a theoretical framework was developed and used to guide the implementation of Healthy Beginning Initiative (HBI) to promote uptake of HIV services in a low-income country. We used the guide for developing a theoretical framework published in Health Promotion Practice. Developing the theoretical framework included seven steps: (1) identifying the essential elements of the intervention; (2) identifying the variables and the context; (3) listing the postulated mechanisms, mediating variables, and postulated outcomes; (4) identifying existing theoretical models supporting the theoretical framework underdevelopment; (5) scripting the theoretical framework into either a figure or sets of statements; (6) conducting content and face validation of the theoretical framework; and (7) revising the theoretical framework. The theoretical framework was developed and used to evaluate HBI's impact on HIV testing, linkage to care and retention in care for pregnant women, their male partners, and newborns. The theoretical framework will also be adapted for other screenings and other settings while remaining true to the essential elements of HBI.


Amblyomma americanum: a potential vector of human ehrlichiosis.
Polymerase chain reaction primers specific for Ehrlichia chaffeensis were used to amplify DNA from extracts of pooled ticks. Amplification was performed on extracts from 140 pools (1,579 total ticks) consisting of three tick genera collected from five states. The characteristic 389-basepair product was observed after amplification of extracts from seven different pools of adult Amblyomma americanum (117 pools, 1,462 ticks), but not from pools of nymphs. No specific product was observed after amplification of 20 pools (105 ticks) of Dermacentor variabilis and three pools of Ixodes scapularis (12 ticks). Ehrlichia chaffeensis was present in A. americanum at a minimum frequency of > or = 0.48%, suggesting that A. americanum may be a vector of human ehrlichiosis.


Modulation of fibronectin gene expression in chondrocytes by viral transformation and substrate attachment.
Chicken vertebral chondrocytes, which normally grow in suspension, synthesize large amounts of cartilage extracellular matrix proteins, but little fibronectin. We have analyzed the effects of both substrate attachment and transformation with a temperature-sensitive mutant of Rous sarcoma virus on fibronectin gene expression in these cells. Our experiments show that viral transformation increases fibronectin synthesis to a greater extent than substrate attachment. Furthermore, transformed chondrocytes have lost the ability to decrease fibronectin synthesis in response to suspension culture, suggesting that transformation alters the normal attachment-responsive control of fibronectin gene expression. Finally, infected substrate-attached chondrocytes shifted to the nonpermissive temperature for transformation use fibronectin RNA more efficiently in protein synthesis than cells grown under the other conditions, suggesting for the first time a role for translational control of fibronectin gene expression.


Therapeutic tumor immunity induced by polyimmunization with melanoma antigens gp100 and TRP-2.
To improve the immunogenicity of melanoma self-antigens, we used a novel strategy of nonviral genetic vaccination coupled with muscle electroporation. Electroporation-enhanced immunization with plasmids encoding either human gp100 or mouse TRP-2 antigens induced only partial rejection of B16 melanoma challenge. However, immunization with a combination of these two antigens caused tumor rejection in 100% of the immunized mice. Splenocytes from combination-immunized animals killed syngeneic targets loaded with peptides derived from both gp100 and TRP-2. Immune cell depletion experiments identified CD8+ T lymphocytes as the primary effectors of antitumor immunity. Most importantly, polyimmunization led to the generation of a therapeutic immune response that significantly improved the mean survival time of mice bearing established lung metastases. These results validated the usefulness of electroporation-enhanced, nonviral genetic immunization for the active immunotherapy of cancer and indicated that using a combination of different tumor antigens may be a decisive strategy for a successful therapeutic vaccination.


Pseudosarcomatous lesions of the urinary bladder, prostate gland, and urethra. A report of three cases and review of the literature.
Three pseudosarcomatous lesions of the genitourinary tract are described. One that arose in the urinary bladder of a 59-year-old man was composed of atypical spindle cells admixed with inflammatory cells in an edematous stroma. The second lesion occurred in the stroma of a hyperplastic prostate gland of a 51-year-old man and was characterized by the presence of numerous atypical mesenchymal cells with large, hyperchromatic nuclei. The third case was that of a 57-year-old woman with a urethral caruncle that contained numerous atypical mesenchymal cells in its stroma. All three patients were treated conservatively, and follow-up for periods ranging from 18 months to eight years has revealed no evidence of recurrence. Knowledge of the various atypical mesenchymal cell proliferations that are encountered in the genitourinary tract may prevent the misdiagnosis of a malignant tumor.


The natural statistics of blur.
Blur from defocus can be both useful and detrimental for visual perception: It can be useful as a source of depth information and detrimental because it degrades image quality. We examined these aspects of blur by measuring the natural statistics of defocus blur across the visual field. Participants wore an eye-and-scene tracker that measured gaze direction, pupil diameter, and scene distances as they performed everyday tasks. We found that blur magnitude increases with increasing eccentricity. There is a vertical gradient in the distances that generate defocus blur: Blur below the fovea is generally due to scene points nearer than fixation; blur above the fovea is mostly due to points farther than fixation. There is no systematic horizontal gradient. Large blurs are generally caused by points farther rather than nearer than fixation. Consistent with the statistics, participants in a perceptual experiment perceived vertical blur gradients as slanted top-back whereas horizontal gradients were perceived equally as left-back and right-back. The tendency for people to see sharp as near and blurred as far is also consistent with the observed statistics. We calculated how many observations will be perceived as unsharp and found that perceptible blur is rare. Finally, we found that eye shape in ground-dwelling animals conforms to that required to put likely distances in best focus.


Case report of schwannoma of the rectum--clinical and pathological contribution.
The case of benign schwannoma of the rectum primarily misdiagnosed as myogenic (neurogenic?) sarcoma is presented. A large tumor of 8 cm in diameter of the anterior rectal wall was removed with wide margins and an artificial anus was constructed. During 12 years of follow-up neither local recurrence nor distant metastases were observed. The patient is still alive and free of the disease. For that reason a surgery specimen of the tumor was pathologically reanalyzed and it showed features of type Antoni A and B tissues. These findings, together with strong reactivity for S-100 protein, Vimentin and negative for Actin supported the diagnosis of benign schwannoma. Because the localization of the tumor in the rectum is extremely rare, clinical and pathological features are presented and discussed.


The fasciclin-like arabinogalactan proteins of Arabidopsis. A multigene family of putative cell adhesion molecules.
Fasciclin-like arabinogalactan proteins (FLAs) are a subclass of arabinogalactan proteins (AGPs) that have, in addition to predicted AGP-like glycosylated regions, putative cell adhesion domains known as fasciclin domains. In other eukaryotes (e.g. fruitfly [Drosophila melanogaster] and humans [Homo sapiens]), fasciclin domain-containing proteins are involved in cell adhesion. There are at least 21 FLAs in the annotated Arabidopsis genome. Despite the deduced proteins having low overall similarity, sequence analysis of the fasciclin domains in Arabidopsis FLAs identified two highly conserved regions that define this motif, suggesting that the cell adhesion function is conserved. We show that FLAs precipitate with beta-glucosyl Yariv reagent, indicating that they share structural characteristics with AGPs. Fourteen of the FLA family members are predicted to be C-terminally substituted with a glycosylphosphatidylinositol anchor, a cleavable form of membrane anchor for proteins, indicating different FLAs may have different developmental roles. Publicly available microarray and expressed sequence tag data were used to select FLAs for further expression analysis. RNA gel blots for a number of FLAs indicate that they are likely to be important during plant development and in response to abiotic stress. FLAs 1,2, and 8 show a rapid decrease in mRNA abundance in response to the phytohormone abscisic acid. Also, the accumulation of FLA1 and FLA2 transcripts differs during callus and shoot development, indicating that the proteins may be significant in the process of competence acquisition and induction of shoot development.


Opposite regulatory effects of cAMP and cGMP on sugar uptake in rat thymocytes.
The present study provides several lines of evidence which indicate that in the rat thymocyte adenosine 3',5'-cyclic monophosphate (cAMP) and guanosine 3',5-cyclic monophosphate (cGMP) induce opposing regulatory effects on 2-deoxyglucose (2-DG) uptake; cAMP is stimulatory, whereas cGMP is inhibitory. First, the cyclic nucleotide analogues dibutyryl cAMP (dBcAMP) and dibutyryl cGMP (dBcGMP) produced a dose-related increase and decrease in thymocyte 2-DG uptake, respectively. Second, 3,5,3'-triiodo-L-thyronine (T3) and epinephrine, which increased cellular cAMP concentration but had no effect on cellular cGMP concentration, increased 2-DG uptake in the rat thymocyte. Third, dBcGMP inhibited the stimulatory effects of dBcAMP, T3, and epinephrine on thymocyte 2-DG uptake. Fourth, prostaglandin E1 and the inhibitors of the cyclic nucleotide phosphodiesterases, 3-isobutyl-1-methylxanthine, theophylline, and caffeine, all increased both cellular cAMP and cGMP concentration but had no effect on 2-DG uptake. Insulin did not change cellular cAMP and cGMP concentration, but produced a dose-related increase in 2-DG uptake by the rat thymocyte. From these results I have concluded that in the rat thymocyte cAMP and cGMP produce opposite effects on sugar uptake and that the effect of certain, but not all, agents on thymocyte sugar uptake results from their modulation of cellular cAMP and cGMP concentration.


A harmonized approach to intracellular cytokine staining gating: Results from an international multiconsortia proficiency panel conducted by the Cancer Immunotherapy Consortium (CIC/CRI).
Previous results from two proficiency panels of intracellular cytokine staining (ICS) from the Cancer Immunotherapy Consortium and panels from the National Institute of Allergy and Infectious Disease and the Association for Cancer Immunotherapy highlight the variability across laboratories in reported % CD8+ or % CD4+ cytokine-positive cells. One of the main causes of interassay variability in flow cytometry-based assays is due to differences in gating strategies between laboratories, which may prohibit the generation of robust results within single centers and across institutions. To study how gating strategies affect the variation in reported results, a gating panel was organized where all participants analyzed the same set of Flow Cytometry Standard (FCS) files from a four-color ICS assay using their own gating protocol (Phase I) and a gating protocol drafted by consensus from the organizers of the panel (Phase II). Focusing on analysis removed donor, assay, and instrument variation, enabling us to quantify the variability caused by gating alone. One hundred ten participating laboratories applied 110 different gating approaches. This led to high variability in the reported percentage of cytokine-positive cells and consequently in response detection in Phase I. However, variability was dramatically reduced when all laboratories used the same gating strategy (Phase II). Proximity of the cytokine gate to the negative population most impacted true-positive and false-positive response detection. Recommendations are provided for the (1) placement of the cytokine-positive gate, (2) identification of CD4+ CD8+ double-positive T cells, (3) placement of lymphocyte gate, (4) inclusion of dim cells, (5) gate uniformity, and 6) proper adjustment of the biexponential scaling.


Cloning, characterization, and high-level expression in Escherichia coli of the Saccharopolyspora erythraea gene encoding an acyl carrier protein potentially involved in fatty acid biosynthesis.
The erythromycin A-producing polyketide synthase from the gram-positive bacterium Saccharopolyspora erythraea (formerly Streptomyces erythraeus) has evident structural similarity to fatty acid synthases, particularly to the multifunctional fatty acid synthases found in eukaryotic cells. Fatty acid synthesis in S. erythraea has previously been proposed to involve a discrete acyl carrier protein (ACP), as in most prokaryotic fatty acid synthases. We have cloned and sequenced the structural gene for this ACP and find that it does encode a discrete small protein. The gene lies immediately adjacent to an open reading frame whose gene product shows sequence homology to known beta-ketoacyl-ACP synthases. A convenient expression system for the S. erythraea ACP was obtained by placing the gene in the expression vector pT7-7 in Escherichia coli. In this system the ACP was efficiently expressed at levels 10 to 20% of total cell protein. The recombinant ACP was active in promoting the synthesis of branched-chain acyl-ACP species by extracts of S. erythraea. Electrospray mass spectrometry is shown to be an excellent method for monitoring the efficiency of in vivo posttranslational modification of ACPs.


Male-specific fruitless specifies the neural substrates of Drosophila courtship behaviour.
Robust innate behaviours are attractive systems for genetically dissecting how environmental cues are perceived and integrated to generate complex behaviours. During courtship, Drosophila males engage in a series of innate, stereotyped behaviours that are coordinated by specific sensory cues. However, little is known about the specific neural substrates mediating this complex behavioural programme. Genetic, developmental and behavioural studies have shown that the fruitless (fru) gene encodes a set of male-specific transcription factors (FruM) that act to establish the potential for courtship in Drosophila. FruM proteins are expressed in approximately 2% of central nervous system neurons, at least one subset of which coordinates the component behaviours of courtship. Here we have inserted the yeast GAL4 gene into the fru locus by homologous recombination and show that (1) FruM is expressed in subsets of all peripheral sensory systems previously implicated in courtship, (2) inhibition of FruM function in olfactory system components reduces olfactory-dependent changes in courtship behaviour, (3) transient inactivation of all FruM-expressing neurons abolishes courtship behaviour, with no other gross changes in general behaviour, and (4) 'masculinization' of FruM-expressing neurons in females is largely sufficient to confer male courtship behaviour. Together, these data demonstrate that FruM proteins specify the neural substrates of male courtship.


Comparative in vitro activity of cefmenoxime, cefotaxime, cefuroxime, cefoxitin, and penicillin against Neisseria gonorrhoeae.
The in vitro activity of a new beta-lactam antibiotic, cefmenoxime, was compared with those of cefotaxime, cofuroxime, cefoxitin, and penicillin against 72 beta-lactamase-negative and 26 beta-lactamase-positive Neisseria gonorrhoeae. Cefmenoxime was as active as cefotaxime and more active than the other three antimicrobial agents. It inhibited all isolates, regardless of beta-lactamase activity, at a concentration of less than or equal to 0.015 microgram/ml.


Trends in Educational Inequalities in Drug Poisoning Mortality: United States, 1994-2010.
We estimated trends in drug poisoning death rates by educational attainment and investigated educational inequalities in drug poisoning mortality by race, gender, and region. We linked drug poisoning death counts from the National Vital Statistics System to population denominators from the Current Population Survey to estimate drug poisoning rates by gender, race, region, and educational attainment (less than high school degree, high school degree, some college, college degree) from 1994 to 2010. There were 372,485 drug poisoning deaths. Education-related inequalities increased during the study among all demographic groups and varied by region. Absolute increases in educational inequalities were higher among Whites than Blacks and men than women. The age-adjusted rate difference between White men with less than a high school degree increased from 8.7 per 100,000 in 1994 to 27.4 in 2010 (change = 18.7). Among Black men, the corresponding increases were 11.7 and 18.3, respectively (change = 6.6). We found strong educational patterning in drug poisoning rates, chiefly by region and race. Rates are highest and increasing the fastest among groups with less education.


The clinical efficacy of citalopram in treatment of emotional disturbances in dementia disorders. A Nordic multicentre study.
In this multicenter study, the clinical efficacy of citalopram was investigated in 98 patients with moderate AD/SDAT or VD using a combined double-blind and open technique with placebo and citalopram. Analyses were made for each diagnosis after four weeks of double-blind treatment. Patients with AD/SDAT treated with citalopram showed a significant improvement in emotional bluntness, confusion, irritability, anxiety, fear/panic, depressed mood and restlessness. Those improvements were not found after treatment with placebo. There were no significant improvements in patients with VD. No improvements were recorded in motor or cognitive impairment. Citalopram provoked few and comparatively mild side-effects. None of the changes observed during the double-blind withdrawal period were identified as withdrawal symptoms or rebound phenomena.


Inhibition of hepatitis C virus replication by pol III-directed overexpression of RNA decoys corresponding to stem-loop structures in the NS5B coding region.
Increasing evidence has shown that the stem-loop (SL) structures in the NS5B coding region of hepatitis C virus (HCV) function as cis-replicating elements that are indispensable for viral replication. We have investigated whether small RNA molecules analogous to the SL structures could inhibit HCV replication. Reporter assays showed that both in vitro transcribed and pol III-directed transcripts corresponding to 5BSL3.1 and 5BSL3.2 efficiently inhibited HCV replicon-encoded luciferase expression. Mutagenesis studies revealed that mutation in 5BSL3.2 which debilitated its binding to NS5B also abolished the ability of 5BSL3.2 RNA to inhibit HCV replication, suggesting that SL RNA inhibits HCV by sequestering the replication complex. Further, adenoviral-mediated expression of the SL RNAs potently blocked the replication of HCV replicon in Huh-7 cells. Importantly, SL RNAs derived from HCV 2a, an evolutionarily distant genotype, were also shown to suppress the replication of HCV 1b replicon in spite of the genetic heterogeneity between the SL elements of the two viruses, implying the potential of SL RNA-based approach to inhibit a wide range of HCV isolates. These results suggest that SL RNA decoys may prove to be useful in the treatment of hepatitis C, which may be advantageous over other sequence-specific gene therapy modalities (such as antisense RNA and siRNA) in preventing the escape of genetic variants.


Emergence of highly-ordered hierarchical nanoscale aggregates on electrostatic binding of self-assembled multivalent (SAMul) cationic micelles with polyanionic heparin.
We report three surfactants, with cationic N,N-di-(3-aminopropyl)-N-methylamine (DAPMA) head groups and aliphatic chains connected via an amide linkage, and investigate their ability to self-assemble and bind polyanionic heparin - a process of potential clinical importance in coagulation control. Modifying the hydrophobic chain length tunes the self-assembly event, with C16-DAPMA having the lowest critical micelle concentration and also being the optimal heparin binder. Remarkably highly structured hierarchical nanoscale aggregates are formed on binding between the spherical cationic micelles and linear polyanionic heparin. C14-DAPMA and C16-DAPMA yield organized polycrystalline assemblies as observed by transmission electron microscopy (TEM), predicted in solution by mesoscale simulations and characterized by small-angle X-ray scattering (SAXS). This confirms that the micelles remain intact during the hierarchical assembly process and become packed in a face-centered cubic manner. The nanoscale assembly formed by C16-DAPMA showed the highest degree of order. Importantly, these studies indicate the impact of hydrophobic modification on self-assembly and heparin binding, demonstrate remarkably high stability of these self-assembled micelles even when forming strong electrostatic interactions with heparin, and provide structural insights into nanoscale hierarchical electrostatic assemblies.


Pearls: sleep disorders.
Sleep disorders and sleep complaints are common in the neurology patient. A good history can often be started with just a few well-worded questions to help identify the major sleep disorders. Identifying sleep-related disorders can help the neurologist broaden the differential diagnosis and treat disorders that can contribute significantly to many comorbidities of other neurologic diseases.


Association Between Transcatheter Aortic Valve Replacement and Subsequent Infective Endocarditis and In-Hospital Death.
Limited data exist on clinical characteristics and outcomes of patients who had infective endocarditis after undergoing transcatheter aortic valve replacement (TAVR). To determine the associated factors, clinical characteristics, and outcomes of patients who had infective endocarditis after TAVR. The Infectious Endocarditis after TAVR International Registry included patients with definite infective endocarditis after TAVR from 47 centers from Europe, North America, and South America between June 2005 and October 2015. Transcatheter aortic valve replacement for incidence of infective endocarditis and infective endocarditis for in-hospital mortality. Infective endocarditis and in-hospital mortality after infective endocarditis. A total of 250 cases of infective endocarditis occurred in 20 006 patients after TAVR (incidence, 1.1% per person-year; 95% CI, 1.1%-1.4%; median age, 80 years; 64% men). Median time from TAVR to infective endocarditis was 5.3 months (interquartile range [IQR], 1.5-13.4 months). The characteristics associated with higher risk of progressing to infective endocarditis after TAVR was younger age (78.9 years vs 81.8 years; hazard ratio [HR], 0.97 per year; 95% CI, 0.94-0.99), male sex (62.0% vs 49.7%; HR, 1.69; 95% CI, 1.13-2.52), diabetes mellitus (41.7% vs 30.0%; HR, 1.52; 95% CI, 1.02-2.29), and moderate to severe aortic regurgitation (22.4% vs 14.7%; HR, 2.05; 95% CI, 1.28-3.28). Health care-associated infective endocarditis was present in 52.8% (95% CI, 46.6%-59.0%) of patients. Enterococci species and Staphylococcus aureus were the most frequently isolated microorganisms (24.6%; 95% CI, 19.1%-30.1% and 23.3%; 95% CI, 17.9%-28.7%, respectively). The in-hospital mortality rate was 36% (95% CI, 30.0%-41.9%; 90 deaths; 160 survivors), and surgery was performed in 14.8% (95% CI, 10.4%-19.2%) of patients during the infective endocarditis episode. In-hospital mortality was associated with a higher logistic EuroSCORE (23.1% vs 18.6%; odds ratio [OR], 1.03 per 1% increase; 95% CI, 1.00-1.05), heart failure (59.3% vs 23.7%; OR, 3.36; 95% CI, 1.74-6.45), and acute kidney injury (67.4% vs 31.6%; OR, 2.70; 95% CI, 1.42-5.11). The 2-year mortality rate was 66.7% (95% CI, 59.0%-74.2%; 132 deaths; 115 survivors). Among patients undergoing TAVR, younger age, male sex, history of diabetes mellitus, and moderate to severe residual aortic regurgitation were significantly associated with an increased risk of infective endocarditis. Patients who developed endocarditis had high rates of in-hospital mortality and 2-year mortality.


Lysis and protoplast formation of group B streptococci by mutanolysin.
Group B streptococci, refractory to previously tested muralysins under physiological conditions, were successfully converted to protoplasts by use of a recently describede N-acetyl muramidase, mutanolysin, derived from a streptomycete. Purified enzyme was effective, but crude preparations, although degrading cell walls, simultaneously produced peculiar effects of cytoplasmic coagulation, retention of cell shape, loss of some intracellular enzymes, and a rise in optical density. Addition of purified mutanolysin to the array of muralysins (group C streptococcal phage-associated lysin, lysozyme), previously successful in preparing protoplasts of different streptococci, now makes possible enzymatic preparation of protoplasts of streptococci of groups A, B, C. D. G, and H.


Tuneable surface shear forces to physically displace nonspecific molecules in protein biomarker detection.
We report a simple method to remove nonspecifically adsorbed species from sensor surface and also improve the detection sensitivity of the sensor using tuneable alternating current (ac) electrohydrodynamics (ac-EHD) forces. These forces generated within few nanometers of an electrode surface (i.e., double layer) engender fluid flow within a serpentine channel containing a long array of the asymmetric electrode pairs, and can easily be tuned externally by changing the frequency and amplitude of the ac-EHD field. Under the optimized experimental conditions, we achieved a 3.5-fold reduction in nonspecific adsorption of non-target proteins with a 1000-fold enhancement in detection sensitivity of the device for the analysis of human epidermal growth factor receptor 2 (HER2) protein spiked in serum. This approach can be applicable in diverse fields including biosensors, cellular and molecular separation systems and biomedical applications to remove/reduce nonspecific adsorption of molecular and cellular species.


An overview of osteoporosis and frailty in the elderly.
Osteoporosis and osteoporotic fractures remain significant public health challenges worldwide. Recently the concept of frailty in relation to osteoporosis in the elderly has been increasingly accepted, with emerging studies measuring frailty as a predictor of osteoporotic fractures. In this overview, we reviewed the relationship between frailty and osteoporosis, described the approaches to measuring the grades of frailty, and presented current studies and future research directions investigating osteoporosis and frailty in the elderly. It is concluded that measuring the grades of frailty in the elderly could assist in the assessment, management and decision-making for osteoporosis and osteoporotic fractures at a clinical research level and at a health care policy level.


Early T cell differentiation lessons from T-cell acute lymphoblastic leukemia.
T cells develop from bone marrow-derived self-renewing hematopoietic stem cells (HSC). Upon entering the thymus, these cells undergo progressive commitment and differentiation driven by the thymic stroma and the pre-T cell receptor (pre-TCR). These processes are disrupted in T-cell acute lymphoblastic leukemia (T-ALL). More than 70% of recurring chromosomal rearrangements in T-ALL activate the expression of oncogenic transcription factors, belonging mostly to three families, basic helix-loop-helix (bHLH), homeobox (HOX), and c-MYB. This prevalence is indicative of their importance in the T lineage, and their dominant mechanisms of transformation. For example, bHLH oncoproteins inhibit E2A and HEB, revealing their tumor suppressor function in the thymus. The induction of T-ALL, nonetheless, requires collaboration with constitutive NOTCH1 signaling and the pre-TCR, as well as loss-of-function mutations for CDKN2A and PTEN. Significantly, NOTCH1, the pre-TCR pathway, and E2A/HEB proteins control critical checkpoints and branchpoints in early thymocyte development whereas several oncogenic transcription factors, HOXA9, c-MYB, SCL, and LYL-1 control HSC self-renewal. Together, these genetic lesions alter key regulatory processes in the cell, favoring self-renewal and subvert the normal control of thymocyte homeostasis.


Extracellular Vesicles from Bone Marrow-Derived Mesenchymal Stem Cells Improve Survival from Lethal Hepatic Failure in Mice.
Stem cell-based therapies have potential for treatment of liver injury by contributing to regenerative responses, through functional tissue replacement or paracrine effects. The release of extracellular vesicles (EV) from cells has been implicated in intercellular communication, and may contribute to beneficial paracrine effects of stem cell-based therapies. Therapeutic effects of bone-marrow derived mesenchymal stem cells (MSC) and vesicles released by these cells were examined in a lethal murine model of hepatic failure induced by d-galactosamine/tumor necrosis factor-α (TNF-α). Systemically administered EV derived from MSC accumulated within the injured liver following systemic administration, reduced hepatic injury, and modulated cytokine expression. Moreover, survival was dramatically increased by EV derived from either murine or human MSC. Similar results were observed with the use of cryopreserved mMSC-EV after 3 months. Y-RNA-1 was identified as a highly enriched noncoding RNA within hMSC-EV compared to cells of origin. Moreover, siRNA mediated knockdown of Y-RNA-1 reduced the protective effects of MSC-EV on TNF-α/ActD-mediated hepatocyte apoptosis in vitro. These data support a critical role for MSC-derived EV in mediating reparative responses following hepatic injury, and provide compelling evidence to support the therapeutic use of MSC-derived EV in fulminant hepatic failure. Stem Cells Translational Medicine 2017;6:1262-1272.


[Characteristics of Silent Alpha Thalassemia Gene in Child-Bearing Adults in Guangdong].
To investigate characteristics of silent alpha thalassemia genes in child-bearing adults in Guangdong, in order to provide data for the prevention and control of hemoglobin H disease. A total of 8 752 cases were collected from January 2016 to December 2020. Gap-PCR was used to detect the deletional of α-thalassemia mutations (-α3.7, -α4.2), while PCR reverse dot blot hybridization assay (RDB) was used to detect the non-deletional α-thalassemia mutations (Hb CS, Hb QS and Hb Westmead). Among 8 752 subjects, 717 cases of silent α-thalassemia were detected, the detection rate was 8.19%, including 555 cases of deletional α-thalassemia (77.41%) and 162 cases of non-deletional α-thalassemia 22.59%. The mean corpuscular volume (MCV) of deletional silent α-thalassemia was (82.09±4.10) fl, and mean corpuscular hemoglobin (MCH) was (27.03±1.37) pg, which both were over the diagnostic cut-off value for thalassemia. The MCV of non-deletional silent α-thalassemia was (81.07±4.93) fl, and MCH was (26.77±2.20) pg. According to the diagnostic criteria, if using MCV<82 fl or (and) MCH<27 pg as a positive criteria for screening thalassemia in the childbearing age, the screening sensitivity was 53.14% and different in different genotype, among which ααQS/αα was 100%, -α3.7/αα, -α4.2/αα, ααCS/αα and ααWS/αα was 62.15%, 63.41%, 44.83% and 39.62%, respectively. Namely, nearly half the carriers of such mutations might have escaped detection as a result of their screening strategy. When a couple is preparing for pregnancy, if one of them has been determined to be mild α-thalassemia or hemoglobin H disease, other half is necessary to carry out silent α thalassemia detection to prevent the birth of children with hemoglobin H disease even if MCV>82 fl and MCH>27 pg.


Probability density function of non-reactive solute concentration in heterogeneous porous formations.
Available models of solute transport in heterogeneous formations lack in providing complete characterization of the predicted concentration. This is a serious drawback especially in risk analysis where confidence intervals and probability of exceeding threshold values are required. Our contribution to fill this gap of knowledge is a probability distribution model for the local concentration of conservative tracers migrating in heterogeneous aquifers. Our model accounts for dilution, mechanical mixing within the sampling volume and spreading due to formation heterogeneity. It is developed by modeling local concentration dynamics with an Ito Stochastic Differential Equation (SDE) that under the hypothesis of statistical stationarity leads to the Beta probability distribution function (pdf) for the solute concentration. This model shows large flexibility in capturing the smoothing effect of the sampling volume and the associated reduction of the probability of exceeding large concentrations. Furthermore, it is fully characterized by the first two moments of the solute concentration, and these are the same pieces of information required for standard geostatistical techniques employing Normal or Log-Normal distributions. Additionally, we show that in the absence of pore-scale dispersion and for point concentrations the pdf model converges to the binary distribution of [Dagan, G., 1982. Stochastic modeling of groundwater flow by unconditional and conditional probabilities, 2, The solute transport. Water Resour. Res. 18 (4), 835-848.], while it approaches the Normal distribution for sampling volumes much larger than the characteristic scale of the aquifer heterogeneity. Furthermore, we demonstrate that the same model with the spatial moments replacing the statistical moments can be applied to estimate the proportion of the plume volume where solute concentrations are above or below critical thresholds. Application of this model to point and vertically averaged bromide concentrations from the first Cape Cod tracer test and to a set of numerical simulations confirms the above findings and for the first time it shows the superiority of the Beta model to both Normal and Log-Normal models in interpreting field data. Furthermore, we show that assuming a-priori that local concentrations are normally or log-normally distributed may result in a severe underestimate of the probability of exceeding large concentrations.


[Intranasal phototherapy for the treatment of allergic rhinitis].
Allergic rhinitis is a frequent disease, accompanied by significant social-economic costs and a negative impact on the quality of life. Phototherapy has a profound immunosuppressive effect and is effectively used in the treatment of several immune mediated skin diseases such as atopic dermatitis. The authors investigated the efficacy of intranasal phototherapy with a combination of low doses of ultraviolet-B, ultraviolet-A and visible light in allergic rhinitis. A randomized, double-blind, placebo-controlled study was conducted in patients with a history of at least 2 years of moderate to severe ragweed-induced allergic rhinitis that was not controlled by anti-allergic drugs. Intranasal phototherapy was performed 3 times a week for 3 weeks. As placebo low intensity visible light was used. Phototherapy resulted in a significant improvement of clinical symptoms for nasal itching, rhinorrhea, sneezing and total nasal score. Scores for nasal obstruction slightly improved during phototherapy while a significant increased was found in the placebo group. In the overall efficacy assessment, both patients and investigators found phototherapy significantly more efficient than placebo. Phototherapy was well tolerated, the only side effect was the slight dryness of the nasal mucosa. These results suggest that intranasal phototherapy is effective for the treatment of allergic rhinitis, and opens up new opportunities for the treatment of immune-mediated mucosal diseases.


Novel interventions targeting on apoptosis and necrosis induced by aluminum chloride in neuroblastoma cells.
Aluminum chloride induces neuroblastoma cell (SH-SY5Y) death following in vitro exposure. The objective of this study is to define apoptosis and necrosis in an in vitro model system of SH-SY5Y cells, and to investigate appropriate defense mechanisms with caspase-3 small interference RNA (siRNA) and necrostatin-1 (Nec-1). SH-SY5Y cells were treated with aluminum chloride for 24 h, followed by analysis of cell death rates and alterations in morphology. The results show that aluminum chloride could induce cell death by a combination of apoptosis and necrosis. Treatment with caspase-3 siRNA resulted in inhibition of caspase-3 gene and protein expression, both indicatives of apoptosis reduction. In addition, decrement of apoptotic rate was evident. Interestingly, treatment with caspase-3 siRNA could markedly up-regulate the expression of LC3- II, indicating a shift of cell death mode, from apoptosis to autophagy. Nec-1 treatment significantly affected necrosis induced by aluminum chloride, resulting in decreased necrotic rates and marked inhibition of LC3- II expression. Results showed for the first time that cell death induced by aluminum chloride could be rescued by caspase-3 siRNA and Nec-1 in SH-SY5Y cells, and co-administration of both produced an additive effect on reducing cell death. These data will pave the way for future studies investigating the prevention of cell death in Al neurotoxicity both in vivo and in vitro.


Increased fluoride uptake and acid resistance by CO2 laser-irradiation through topically applied fluoride on human enamel in vitro.
The aim of the current in vitro study was to evaluate the effect of CO(2)-laser treatment immediately after applying amine fluoride solution on enamel. It was hypothesized that such a treatment would increase enamel fluoride uptake, and reduce dissolution rate and thermal surface alterations. Fluoride uptake was determined in 40 human enamel sections randomly assigned to four groups (n=10), which were either left untreated (1), exposed to a 1% amine fluoride solution for 15s without irradiation (2), irradiated for 15s with a continuous-wave carbon dioxide laser (3), or laser-treated for 15s through the amine fluoride solution applied immediately beforehand (4). Fluoride uptake was determined with an ion selective electrode after acid dissolution of the specimens (surface and subsurface layers). For the determination of acid resistance, another 40 enamel sections were treated according to the above protocol. Acid resistance was determined in surface and subsurface layers by measuring eluted calcium upon 3% lactic acid exposure with atomic absorption spectrometry. Enamel surface alterations after laser irradiation were monitored using scanning electron microscopy. Laser irradiation through the fluoride solution led to significantly higher fluoride contents in the surface enamel layer compared to fluoride treatment alone or laser treatment alone (p=0.002). Laser treatment with or without fluoride resulted in an increased acid resistance of enamel specimens. Fewer surface alterations were observed upon SEM examination of specimens irradiated through the amine fluoride solution compared to counterparts treated with laser only. CO(2) laser light application through an amine fluoride solution may be useful and effective in the prevention of caries.


Prevention of adolescent substance abuse through social skill development.
The purpose of this chapter is to evaluate the capability of social skills training approaches for preventing adolescent substance abuse and to suggest directions for future research in this area. Social skills and training are defined from the perspective of social learning theory. Training approaches are then reviewed for their effect on drug use and drug use-related behaviors. The results indicate that social skills training reduces substance use and related behaviors such as aggression, withdrawal, truancy, and stealing in adolescents. Outcome was enhanced by the inclusion of modeling in a training program and the use of normal (versus problem or disordered) adolescents. In addition, discriminant analyses indicated that the more positive outcomes were discriminable by a focus on prevention, experimental rigor, and training comprehensiveness, that is, inclusion of multiple techniques and skills. Follow-up results showed that training effects were maintained, and that they could be generalized to several areas of everyday adolescent functioning. The findings suggest that social skills training is an effective means for substance abuse prevention in adolescents, and that it may have an advantage over more direct prevention methods in terms of long-term generalization capabilities. Finally, several issues are raised for their potential impact on social skills training as a strategy for substance abuse prevention in adolescents. These issues encompass conceptual, methodological, and evaluative domains: development of a taxonomy of person-situation specificity characteristics to increase the validity and reliability of assessment and training procedures; assessment of the relative contribution of individual differences in adolescents to training outcome; identification of qualitative developmental shifts in social skills as adolescents progress through grade, school, and family transitions; and comparison of the maintenance and generalization effects, over time, of social skills training with more direct abuse prevention methods. Guidelines are offered for addressing these issues, as well as directions for the role of social skills training research in future drug abuse prevention efforts.