Dissipation kinetics, residue determination and consumer risk assessment of acephate on green pea by gas chromatography-electron capture detector.
The present study on "acephate persistence on green pea" was conducted in SKUAST-Kashmir. The study aimed to determine the persistence, dissipation kinetics and waiting period of acephate on green pea. Acephate was sprayed at 75% soluble powder (SP) at 560 g a.i.ha-1 at the fruiting stage followed by another application at a 10 day interval. A rapid and accurate method (quick, easy, cheap, effective, rugged and safe, QuEChERS) was used for extraction and the residue was determined by gas chromatography-electron capture detection on a CPSIL-8CB capillary column (0.25um film thickness, 0.25 mm i.d, 30 m length). At the fortification levels of 0.05, 0.1 and 0.5 mg kg-1 , the percentage recovery of acephate on green pea was found in the range of 71-107%. The initial deposit of green pea was estimated to be 0.37 mg kg-1 . At the indicated dose, the residue of acephate on green pea dissipated below the limit of quantification of 0.05 mg kg-1 after 10 days. Acephate degradation was quick in green pea, with a half-life of 4.07 days. For safe eating of green peas, a 10 day waiting period is recommended. The gas chromatography-electron capture detection technique was validated by following the SANTE standards.


RAS signalling is abnormal in a c-raf1 MEK1 double mutant.
A mutant rat cell clone that suppresses the transformation defects of RAS effector loop substitutions is heterozygous for mutations in c-raf1 and MEK1. The mutant cells can be transformed by many otherwise defective RAS effector mutants, including RAS genes with the effector regions of distantly related GTPases, even though the encoded RAS proteins do not interact with either the mutant or wild-type RAF in Saccharomyces cerevisiae. While the significance of the c-raf1 mutation is unclear, the MEK1 mutation increases MEK1 activity and leads to activation of mitogen-activated protein kinase. The mutant MEK1 is coupled to the epidermal growth factor pathway but exhibits decreased physical interaction with RAF. When overexpressed, the MEK1 mutation is transforming and causes hyperphosphorylation of RAF. Signalling from RAS to MEK1 may be mediated by something other than RAF alone, but signalling through MEK1 is probably sufficient for RAS transformation.


Current knowledge and pattern of use of family planning methods among a severely ill female Nigerian psychiatric outpatients: implication for existing service.
Despite the acknowledged benefits accruable to the mentally ill from the use of contraceptives, research and dedicated reproductive health services for this group of people in Nigeria are still underdeveloped or non-existent in many cases. This study aims at identifying the current status of knowledge and pattern of use of modern contraceptives in a clinic population. A cross-sectional descriptive study of 100 women with severe mental illness attending psychiatric outpatient clinic in Ilorin, Nigeria, using a semi-structured questionnaire. A majority (88%) of the women had good knowledge of family planning, many (61%) were interested in its use but at least half of them had not used any method and current use was just 27%. Attitude to family planning was generally positive as indicated by desired family size (71%) of less than 4 children with 6 out of 10 desiring future intention to use. The male condom was the most known and ever used method (68%, 37%), respectively. Most (81%) of these women desired that family planning information be provided at the psychiatric clinic but fewer of them discussed this with their partners. Considering discrepancies between knowledge, interest and use, the potential health benefits of fertility regulation, and the adverse effects of unregulated family, and contraceptive use is still relatively low. There is need for improvement which might only be possible if psychiatric focused programs are initiated. The large number of women who desired clinic-based family planning education suggests that the introduction of such program is feasible.


Increased Ascorbate Content of Glioblastoma Is Associated With a Suppressed Hypoxic Response and Improved Patient Survival.
Glioblastoma multiforme is a challenging disease with limited treatment options and poor survival. Glioblastoma tumours are characterised by hypoxia that activates the hypoxia inducible factor (HIF) pathway and controls a myriad of genes that drive cancer progression. HIF transcription factors are regulated at the post-translation level via HIF-hydroxylases. These hydroxylases require oxygen and 2-oxoglutarate as substrates, and ferrous iron and ascorbate as cofactors. In this retrospective observational study, we aimed to determine whether ascorbate played a role in the hypoxic response of glioblastoma, and whether this affected patient outcome. We measured the ascorbate content and members of the HIF-pathway of clinical glioblastoma samples, and assessed their association with clinicopathological features and patient survival. In 37 samples (37 patients), median ascorbate content was 7.6 μg ascorbate/100 mg tissue, range 0.8 - 20.4 μg ascorbate/100 mg tissue. In tumours with above median ascorbate content, HIF-pathway activity as a whole was significantly suppressed (p = 0.005), and several members of the pathway showed decreased expression (carbonic anhydrase-9 and glucose transporter-1, both p < 0.01). Patients with either lower tumour HIF-pathway activity or higher tumour ascorbate content survived significantly longer than patients with higher HIF-pathway or lower ascorbate levels (p = 0.011, p = 0.043, respectively). Median survival for the low HIF-pathway score group was 362 days compared to 203 days for the high HIF-pathway score group, and median survival for the above median ascorbate group was 390 days, compared to the below median ascorbate group with 219 days. The apparent survival advantage associated with higher tumour ascorbate was more prominent for the first 8 months following surgery. These associations are promising, suggesting an important role for ascorbate-regulated HIF-pathway activity in glioblastoma that may impact on patient survival.


DNA polymorphisms in two paraoxonase genes (PON1 and PON2) are associated with the risk of coronary heart disease.
A common polymorphism at codon 192 in the human paraoxonase (PON) 1 gene has been shown to be associated with increased risk for coronary heart disease (CHD) in Caucasian populations. However, these findings have not been reported consistently in all Caucasian and non-Caucasian populations, suggesting that this is not a functional mutation but may mark a functional mutation present in either PON1 or a nearby gene. Recently, two other PON-like genes, designated "PON2" and "PON3," have been identified, and they are linked with the known PON1 gene on chromosome 7. Identification of additional polymorphisms in the PON-gene cluster may help to locate the functional polymorphism. In this report, we describe the existence of a common polymorphism at codon 311 (Cys-->Ser; PON2*S) in the PON2 gene, as well as its association with CHD alone and in combination with the PON1 codon 192 polymorphism in Asian Indians. The frequency of the PON2*S allele was significantly higher in cases than in controls (.71 vs. .61; P=.016). The age- and sex-adjusted odds ratio (OR) was 2.5 (95% confidence interval &sqbl0;95% CI&sqbr0;=1.8-3.1; P=.0090) for the PON2*S allele carriers. Further stratification of the PON2*S association, on the basis of the presence or absence of the PON1*B allele, showed that the CHD risk associated with the PON2*S allele was confined to PON1*B-allele carriers. Likewise, the PON1*B-allele risk was present only among PON2*S carriers. Age- and sex-adjusted ORs for the PON2*S and PON1*B were 3.6 (95% CI=2.6-4.6; P=.011) and 2.9 (95% CI=2.4-3.5; P=.0002) among the PON1*B and PON2*S carriers, respectively. Our data indicate that both polymorphisms synergistically contribute to the CHD risk in this sample and that this genetic risk is independent of the conventional plasma lipid profile.


Crystal structure of Escherichia coli pyruvate kinase type I: molecular basis of the allosteric transition.
Pyruvate kinase (PK) plays a major role in the regulation of glycolysis. Its catalytic activity is controlled by the substrate phosphoenolpyruvate and by one or more allosteric effectors. The crystal structures of the non-allosteric PKs from cat and rabbit muscle are known. We have determined the three-dimensional structure of the allosteric type I PK from Escherichia coli, in order to study the mechanism of allosteric regulation. The 2.5 A resolution crystal structure of the unligated type I PK in the inactive T-state shows that each subunit of the homotetrameric enzyme comprises a (beta/alpha)8-barrel domain, a flexible beta-barrel domain and a C-terminal domain. The allosteric and active sites are located at the domain interfaces. Comparison of the T-state E. coli PK with the non-allosteric muscle enzyme, which is thought to adopt a conformation similar to the active R-state, reveals differences in the orientations of the beta-barrel and C-terminal domains of each subunit, which are rotated by 17 degrees and 15 degrees, respectively. Moreover, the relative orientation of the four subunits differs by about 16 degrees in the two enzymes. Highly conserved residues at the subunit interfaces couple these movements to conformational changes in the substrate and allosteric effector binding sites. The subunit rotations observed in the T-state PK induce a shift in loop 6 of the (beta/alpha)8-barrel domain, leading to a distortion of the phosphoenolpyruvate-binding site accounting for the low substrate affinity of the T-state enzyme. Our results suggest that allosteric control of PK is accomplished through remarkable domain and subunit rotations. On transition from the T- to the R-state all 12 domains of the functional tetramer modify their relative orientations. These concerted motions are the molecular basis of the coupling between the active centre and the allosteric site.


A model in mice for the study of the early death phenomenon after vaccination and challenge with rabies virus.
The reactions of mice were studied for a period of 10 days after their vaccination with an inactivated rabies vaccine. The kinetics of their resistance to an intracerebral challenge and neutralizing antibody activity of their serum were determined daily. Protection began on the fourth day after vaccination and was approximately correlated with virus-neutralizing antibody titres from the sixth to the tenth day. However, in vaccinated mice still unprotected, death following the intracerebral challenge occurred earlier than in unvaccinated control mice. This "early death phenomenon" is proposed as a model for immuno-pathological study of reaction to rabies vaccines.


[The investigation of accidental swallowing of Fluoride toothpaste in young children]
OBJECTIVE:This study was to investigate the accidental swallowing of fluoride toothpastes in young children during their toothbrushing and then to discuss the security of fluoride toothpastes used by young children.METHODS:The accidental ingesting of fluoride toothpastes in 180 young children aged 3-6 years old was assayed with fluoride-specific electrode.RESULTS:Among the 180 children invited to join the test,the average amount of the ingestion of fluoride toothpastes was 0.053g.The average swallowing ratio was 26.163%.The maximums presented in the group aged 3.0 to 3.9years old were 0.082g and 30.348%;respectively.CONCLUSION:Young children swallow a quantity of toothpaste on the brush during toothbrushing inevitably.It may result in the excessive daily fluoride ingestion which includes the fluoride ingestion from toothpaste,food and surroundings.So the incidence of fluorosis will increase certainly.


The epidemiology of myasthenia gravis.
Population-based studies of the epidemiology of myasthenia gravis (MG) have been conducted for over 50 years. Over that time, there has been a clear trend towards an increase in the reported prevalence of the disease. In recent years, there has also been an interest in determining a reasonably accurate estimate of the number of MG patients in the United States. Current estimates place the prevalence at a high value of about 20 per 100,000. The year 2000 U.S. population estimate is slightly less than 280 million. A crude estimate of the number of MG patients derived from the population estimate and the reported prevalence from selected populations indicates that there are between 53,000 and 59,900 patients in the United States at this time. The age and ethnic distributions in the United States are evolving from those that were present when the majority of the population-based studies were done, and the distribution of severity of the disease may also have changed. Future studies of the epidemiology of MG should take these factors into account, and further research into the economic and quality-of-life impact of the disease in the population is needed.


Natural history of bronchial preinvasive lesions.
Preinvasive bronchial lesions defined as dysplasia and carcinoma in situ (CIS) have been considered as precursors of squamous cell carcinoma of the lung. The risk and rate of progression of preinvasive lesions to invasive squamous cell carcinoma as well as the mechanism of progression or regression are incompletely understood. While the evidence for the multistage, stepwise progression model is weak with relatively few documented lesions that progress through various grades of dysplasia to CIS and then to invasive carcinoma, the concept of field carcinogenesis is strongly supported. The presence of high-grade dysplasia or CIS is a risk marker for lung cancer both in the central airways and peripheral lung. Genetic alterations such as loss of heterozygosity in chromosome 3p or chromosomal aneusomy as well as host factors such as the inflammatory load and levels of anti-inflammatory proteins in the lung influence the progression or regression of preinvasive lesions. CIS is different than severe dysplasia at the molecular level and has different clinical outcome. Molecular analysis of dysplastic lesions that progress to CIS or invasive cancer and rare lesions that progress rapidly from hyperplasia or metaplasia to CIS or invasive cancer will shed light on the key molecular determinants driving development to an invasive phenotype versus those associated with tobacco smoke damage.


Calbindin-D28K is increased in the ventral horn of spinal cord by neuroprotective factors for motor neurons.
Slow glutamate-mediated neuronal degeneration is implicated in the pathophysiology of motor neuron diseases such as amyotrophic lateral sclerosis (ALS). The calcium-binding proteins calbindin-D28K and parvalbumin have been reported to protect neurons against excitotoxic insults. Expression of calbindin-D28K is low in adult human motor neurons, and vulnerable motor neurons additionally may lack parvalbumin. Thus, it has been speculated that the lack of calcium-binding proteins may, in part, be responsible for early degeneration of the population of motor neurons most vulnerable in ALS. Using a rat organotypic spinal cord slice system, we examined whether the most potent neuroprotective factors for motor neurons can increase the expression of calbindin-D28K or parvalbumin proteins in the postnatal spinal cord. After 4 weeks of incubation of spinal cord slices with 1) glial cell line-derived neurotrophic factor (GDNF), 2) neurturin, 3) insulin-like growth factor I (IGF-I), or 4) pigment epithelium-derived factor (PEDF), the number of calbindin-D28K -immunopositive large neurons (>20 μm) in the ventral horn was higher under the first three conditions, but not after PEDF, compared with untreated controls. Under the same conditions, parvalbumin was not upregulated by any neuroprotective factor. The same calbindin increase was true of IGF-I and GDNF in a parallel glutamate toxicity model of motor neuron degeneration. Taken together with our previous reports from the same model, which showed that all these neurotrophic factors can potently protect motor neurons from slow glutamate injury, the data here suggest that upregulation of calbindin-D28K by some of these factors may be one mechanism by which motor neurons can be protected from glutamate-induced, calcium-mediated excitotoxicity.


Sleep Spindle Characteristics in Obstructive Sleep Apnea Syndrome (OSAS).
Background: We compared the density and duration of sleep spindles topographically in stage 2 and 3 of non-rapid eye movement sleep (N2 and N3) among adults diagnosed with Obstructive Sleep Apnea Syndrome (OSAS) and healthy controls. Materials and Methods: Thirty-one individuals with OSAS (mean age: 48.50 years) and 23 healthy controls took part in the study. All participants underwent a whole night polysomnography. Additionally, those with OSAS were divided into mild, moderate and severe cases of OSAS. Results: For N2, sleep spindle density did not significantly differ between participants with and without OSAS, or among those with mild, moderate and severe OSAS. For N3, post-hoc analyses revealed significantly higher spindle densities in healthy controls and individuals with mild OSAS than in those with moderate or severe OSAS. Last, in N2 a higher AHI was associated with a shorter sleep spindle duration. Conclusion: OSAS is associated with a significantly lower spindle density in N3 and a shorter spindle duration in N2. Our results also revealed that, in contrast to moderate and severe OSAS, the sleep spindle characteristics of individuals with mild OSAS were very similar to those of healthy controls.


Planted matching problems on random hypergraphs.
We consider the problem of inferring a matching hidden in a weighted random k-hypergraph. We assume that the hyperedges' weights are random and distributed according to two different densities conditioning on the fact that they belong to the hidden matching or not. We show that for k>2 and in the large-graph-size limit, an algorithmic first-order transition in the signal strength separates a regime in which a complete recovery of the hidden matching is feasible from a regime in which partial recovery is possible. This is in contrast to the k=2 case, where the transition is known to be continuous. Finally, we consider the case of graphs presenting a mixture of edges and 3-hyperedges, interpolating between the k=2 and the k=3 cases, and we study how the transition changes from continuous to first order by tuning the relative amount of edges and hyperedges.


[Intestinal parasite infections in a periurban community from the Province of Buenos Aires, Argentina].
A survey for intestinal parasites was performed on 38 individuals within the urban area of La Plata City (Province of Buenos Aires, Argentina). This community is composed of brick-factory workers who also live in the factory premises. An analysis for intestinal parasites was done on fecal samples collected serially and by means of anal swabs and thereafter preserved in formol solution. At the same time, the occurrence of the parasites under study as well as that of commensal organisms was investigated in water and soil samples within the factory environs. Information was also obtained from the members of this community as their age, sex, birthplace, and recent travels, either abroad or to the interior of Argentina. The prevalence of the pathogens and commensal parasites was 89.5%. The frequency of protozoans and helminths was: G. lamblia 26.3%, B. hominis 65.8%, A. lumbricoides 7.9%, H. nana 2.6%, Uncinaria sp. 7.9%, S. stercoralis 2.6% and E. vermicularis 42.1%. None of these parasite or commensal organisms were present in the water samples investigated. Four out of 20 soil samples analyzed contained parasitic elements: T. canis eggs (one), G. lamblia cysts (two) and A. lumbricoides eggs (one). These results indicated that most important factors causing such a high prevalence of coproparasites were the poor conditions of personal and community hygiene in combination with the frequent travels to the north and the northeast of the country, regions which are endemic parasitic areas. The implementation of programs on health education and communal sanitation would contribute in the control of this health problem.


S-nitrosylation of HDAC2 regulates the expression of the chromatin-remodeling factor Brm during radial neuron migration.
Dynamic epigenetic modifications play a key role in mediating the expression of genes required for neuronal development. We previously identified nitric oxide (NO) as a signaling molecule that mediates S-nitrosylation of histone deacetylase 2 (HDAC2) and epigenetic changes in neurons. Here, we show that HDAC2 nitrosylation regulates neuronal radial migration during cortical development. Bead-array analysis performed in the developing cortex revealed that brahma (Brm), a subunit of the ATP-dependent chromatin-remodeling complex BRG/brahma-associated factor, is one of the genes regulated by S-nitrosylation of HDAC2. In the cortex, expression of a mutant form of HDAC2 that cannot be nitrosylated dramatically inhibits Brm expression. Our study identifies NO and HDAC2 nitrosylation as part of a signaling pathway that regulates cortical development and the expression of Brm in neurons.


Multiple stressors interact primarily through antagonism to drive changes in the coral microbiome.
Perturbations in natural systems generally are the combination of multiple interactions among individual stressors. However, methods to interpret the effects of interacting stressors remain challenging and are biased to identifying synergies which are prioritized in conservation. Therefore we conducted a multiple stressor experiment (no stress, single, double, triple) on the coral Pocillopora meandrina to evaluate how its microbiome changes compositionally with increasing levels of perturbation. We found that effects of nutrient enrichment, simulated predation, and increased temperature are antagonistic, rather than synergistic or additive, for a variety of microbial community diversity measures. Importantly, high temperature and scarring alone had the greatest effect on changing microbial community composition and diversity. Using differential abundance analysis, we found that the main effects of stressors increased the abundance of opportunistic taxa, and two-way interactions among stressors acted antagonistically on this increase, while three-way interactions acted synergistically. These data suggest that: (1) multiple statistical analyses should be conducted for a complete assessment of microbial community dynamics, (2) for some statistical metrics multiple stressors do not necessarily increase the disruption of microbiomes over single stressors in this coral species, and (3) the observed stressor-induced community dysbiosis is characterized by a proliferation of opportunists rather than a depletion of a proposed coral symbiont of the genus Endozoicomonas.


Methods for the determination of androgen receptor content in human prostatic tissue.
Human prostatic androgen receptor content can be measured reliably in either fresh or bulk tissue stored in liquid nitrogen using (3H) R 1881 at incubation conditions of 4 C for 20 hr. Powdered tissue stored in liquid nitrogen for more than 12 days shows a marked deterioration in receptor content. Although multiple point dextran coated charcoal assays analyzed by Scatchard plot are preferable for receptor quantitation of bulk tissue, the single saturating dose assay provides useful information on needle biopsy specimens. When this technique is used to evaluate samples with protein concentrations less than 1 mg per ml, the use of hydroxylapatite to separate receptor bound and free steroid is superior to the use of dextran coated charcoal. The addition of sodium molybdate to the homogenization buffers results in a marked increase in cystosolic androgen receptor content and a decrease in extractable nuclear receptor content. The use of a vertical rotor to ensure short centrifugation times enhances the reliability of sucrose density gradient analyses of human prostatic androgen receptor.


Planning for resilience: Incorporating scenario and model uncertainty and trade-offs when prioritizing management of climate refugia.
Climate change has become the greatest threat to the world's ecosystems. Locating and managing areas that contribute to the survival of key species under climate change is critical for the persistence of ecosystems in the future. Here, we identify 'Climate Priority' sites as coral reefs exposed to relatively low levels of climate stress that will be more likely to persist in the future. We present the first analysis of uncertainty in climate change scenarios and models, along with multiple objectives, in a marine spatial planning exercise and offer a comprehensive approach to incorporating uncertainty and trade-offs in any ecosystem. We first described each site using environmental characteristics that are associated with a higher chance of persistence (larval connectivity, hurricane influence, and acute and chronic temperature conditions in the past and the future). Future temperature increases were assessed using downscaled data under four different climate scenarios (SSP1 2.6, SSP2 4.5, SSP3 7.0 and SSP5 8.5) and 57 model runs. We then prioritized sites for intervention (conservation, improved management or restoration) using robust decision-making approaches that select sites that will have a benign climate under most climate scenarios and models. The modelling work is novel because it solves two important issues. (1) It considers trade-offs between multiple planning objectives explicitly through Pareto analyses and (2) It makes use of all the uncertainty around future climate change. Priority intervention sites identified by the model were verified and refined through local stakeholder engagement including assessments of local threats, ecological conditions and government priorities. The workflow is presented for the Insular Caribbean and Florida, and at the national level for Cuba, Jamaica, Dominican Republic and Haiti. Our approach allows managers to consider uncertainty and multiple objectives for climate-smart spatial management in coral reefs or any ecosystem across the globe.


Comparison of IOL power calculation methods and intraoperative wavefront aberrometer in eyes after refractive surgery.
To compare preoperative methods for calculating intraocular lens (IOL) power versus the intraoperative wavefront aberrometer in eyes with a history of refractive surgery. A retrospective study of 46 eyes (33 patients) with previous refractive surgery that underwent subsequent cataract surgery was conducted. Suggested IOL power predicted by ORange intraoperative wavefront aberrometer (WaveTec Vision Systems, Inc., Aliso Viejo, CA) was compared to power predicted by the (1) SRK-T formula using keratometry and axial length measurements from the IOLMaster (Carl Zeiss Meditec, Dublin, CA), (2) average central keratometry (Avg K) from corneal topography, and (3) average IOL power predicted by the American Society of Cataract and Refractive Surgery (ASCRS) web site. No historical information was used for the calculations. IOL power required for emmetropia was back-calculated using manifest refraction and implanted IOL power after cataract surgery. Mean age was 60 ± 7.9 years. Fifteen percent had a history of myopic photorefractive keratectomy (n = 7), 57% myopic LASIK (n = 26), 13% hyperopic LASIK (n = 6), and 22% radial keratectomy (RK) (n = 10). In 37% of cases, ORange predicted IOL power to within ±0.50 diopters (D) of emmetropia, compared to 30% for IOLMaster keratometry, 26% for Avg K, and 17% for ASCRS web site. In eyes after myopic treatment, ORange, IOLMaster, Avg K, and ASCRS web site predicted within ±0.50 D of emmetropia in 39%, 27%, 24%, and 18%, respectively, and within ±1.0 D in 60%, 39%, 39%, and 51%, respectively. In eyes after RK, ORange, Avg K, and ASCRS web site predicted to within ±0.50 D of emmetropia in 14% and the IOLMaster in 43% cases. Although the ORange most often predicted to within ±0.5 D of emmetropia, no method was able to achieve this accuracy more than 50% of the time. Predictions for eyes after RK were worse than for other types of refractive procedures.


Ammonium-acetate is sensed by gustatory and olfactory neurons in Caenorhabditis elegans.
Caenorhabditis elegans chemosensation has been successfully studied using behavioral assays that treat detection of volatile and water soluble chemicals as separate senses, analogous to smell and taste. However, considerable ambiguity has been associated with the attractive properties of the compound ammonium-acetate (NH(4)Ac). NH(4)Ac has been used in behavioral assays both as a chemosensory neutral compound and as an attractant. Here we show that over a range of concentrations NH(4)Ac can be detected both as a water soluble attractant and as an odorant, and that ammonia and acetic acid individually act as olfactory attractants. We use genetic analysis to show that NaCl and NH(4)Ac sensation are mediated by separate pathways and that ammonium sensation depends on the cyclic nucleotide gated ion channel TAX-2/TAX-4, but acetate sensation does not. Furthermore we show that sodium-acetate (NaAc) and ammonium-chloride (NH(4)Cl) are not detected as Na(+) and Cl(-) specific stimuli, respectively. These findings clarify the behavioral response of C. elegans to NH(4)Ac. The results should have an impact on the design and interpretation of chemosensory experiments studying detection and adaptation to soluble compounds in the nematode Caenorhabditis elegans.


Double photoionization and dication fragmentation of CF3I: Experiment and theory.
The double photoionization of CF(3)I and the electronic structure and the dissociation dynamics of the CF(3)I(++) dication have been investigated using large ab initio calculations and coincidence techniques. The double photoionization spectrum of CF(3)I consists of a continuous background with a number of narrow bands superimposed. The spectrum is attributed here to the population of groups of close lying electronic states interacting mutually by spin-orbit, spin-spin, and rovibronic couplings. At energies near the vertical double ionization threshold, CF(3) (+)+I(+) ionic fragments are produced. At higher energies, a very specific dissociation with double charge retained on one fragment, CF(3)I(++)-->CF(2)I(++)+F becomes dominant and is attributed to a specific group of dication electronic states.


Long-term efficacy and toxicity of cholinesterase inhibitors in the treatment of Alzheimer disease.
Though the symptoms of Alzheimer disease go on for years, the phase 3 trials of the cholinesterase inhibitors (ChEIs), the current mainstay of symptomatic pharmacotherapy for this condition, were typically of only 3- to 6-months' duration. We have limited data on long-term (that is, a year or more) therapy with these agents. In this review, we explore the available information on the biological and clinical effects of long-term ChEI therapy, what happens when these agents are discontinued, and examine what others have recommended An individualized approach to deciding on whether to carry on with a ChEI should be taken. If continued, treatment goals should be clarified and patients monitored over time, for both drug-related benefits and adverse effects.


Enhancing the Biopharmacological Characteristics of Asperosaponin VI: Unveiling Dynamic Self-Assembly Phase Transitions in the Gastrointestinal Environment.
Asperosaponin VI (ASP VI) as an active ingredient of Dipsacus asperoides, which has a wide range of biological and pharmacological activity. However, its development and application are restricted due to the poor gastrointestinal permeability and oral bioavailability. This investigation aims to reveal the influence of the self-assembled structure by the interaction between ASP VI and endogenous components NaTC and/or DOPC in the gastrointestinal environment on its biopharmaceutical properties, and novelty elucidated the molecular mechanism for the formation of self-assembled nanomicelles. This change in phase state in gastrointestinal fluids is characterized by dynamic light scattering (DLS) and transmission electron microscope (TEM). UPLC-Q-TOF-MS was used to analyze the composition of phase components and the exposure of nanomicelles in vivo. Molecular dynamics simulation (MDS) was applied to preliminarily elucidate the self-assembly mechanism of ASP VI in the gastrointestinal environment. Furthermore, theS8 promoting absorption mechanism of nanomicelles were investigated through in vivo pharmacokinetic experiments, parallel artificial membrane permeability assay (PAMPA), quadruple single-pass intestinal perfusion in rats, and Caco-2 cell monolayer model. We demonstrated that the ASP VI could spontaneously form dynamic self-assembled structures with sodium taurocholate (NaTC) and dipalmitoyl phosphatidylcholine (DOPC) during gastrointestinal solubilization, which promoted the gastrointestinal absorption and permeability of ASP VI and increased its exposure in vivo, thus improving the biopharmacological characteristics of ASP VI. Moreover, ASP VI-NaTC-DOPC-self-assembled nanostructures (ASP VI-NaTC-DOPC-SAN) manifested higher cellular uptake in Caco-2 cells as evidenced by flow cytometry and confocal microscopy, and this study also preliminarily revealed the mechanism of self-assembly formation of ASP VI with endogenous components NaTC and DOPC driven by electrostatic and hydrogen bonding interactions. This study provides evidence that the dynamic self-assembled phase transition may play a key role in improving the biopharmacological characteristics of insoluble or low permeability active ingredients during the gastrointestinal dissolution of Chinese medicines.


Effects of positive pressure breathing on right and left ventricular preload and afterload.
Positive pressure ventilation with positive end-expiratory pressure (PEEP) is used to treat patients with pulmonary edema. PEEP usually raises arterial oxygen tension, but also may reduce cardiac output and stroke volume. The fall in cardiac output could be due to a fall in preload, an increase in afterload, or a decrease in myocardial contractility of either ventricle. Right ventricular afterload increases during PEEP but not enough to cause cardiac output to fall. Right atrial transmural pressure increases; right ventricular diastolic shape changes with an increase in septal to lateral diameter. Left ventricular afterload and end-diastolic volume fall during PEEP even though left atrial transmural pressure is maintained; an exaggerated decrease in left ventricular diastolic septal to lateral diameter suggests that a septal shift impairs left ventricular end-diastolic volume.


The regularity of sustained firing reveals two populations of slowly adapting touch receptors in mouse hairy skin.
Touch is initiated by diverse somatosensory afferents that innervate the skin. The ability to manipulate and classify receptor subtypes is prerequisite for elucidating sensory mechanisms. Merkel cell-neurite complexes, which distinguish shapes and textures, are experimentally tractable mammalian touch receptors that mediate slowly adapting type I (SAI) responses. The assessment of SAI function in mutant mice has been hindered because previous studies did not distinguish SAI responses from slowly adapting type II (SAII) responses, which are thought to arise from different end organs, such as Ruffini endings. Thus we sought methods to discriminate these afferent types. We developed an epidermis-up ex vivo skin-nerve chamber to record action potentials from afferents while imaging Merkel cells in intact receptive fields. Using model-based cluster analysis, we found that two types of slowly adapting receptors were readily distinguished based on the regularity of touch-evoked firing patterns. We identified these clusters as SAI (coefficient of variation = 0.78 +/- 0.09) and SAII responses (0.21 +/- 0.09). The identity of SAI afferents was confirmed by recording from transgenic mice with green fluorescent protein-expressing Merkel cells. SAI receptive fields always contained fluorescent Merkel cells (n = 10), whereas SAII receptive fields lacked these cells (n = 5). Consistent with reports from other vertebrates, mouse SAI and SAII responses arise from afferents exhibiting similar conduction velocities, receptive field sizes, mechanical thresholds, and firing rates. These results demonstrate that mice, like other vertebrates, have two classes of slowly adapting light-touch receptors, identify a simple method to distinguish these populations, and extend the utility of skin-nerve recordings for genetic dissection of touch receptor mechanisms.


Clinical Outcomes After Anterior Shoulder Stabilization in Overhead Athletes: An Analysis of the MOON Shoulder Instability Consortium.
Traumatic anterior shoulder instability is a common condition affecting sports participation among young athletes. Clinical outcomes after surgical management may vary according to patient activity level and sport involvement. Overhead athletes may experience a higher rate of recurrent instability and difficulty returning to sport postoperatively with limited previous literature to guide treatment. To report the clinical outcomes of patients undergoing primary arthroscopic anterior shoulder stabilization within the Multicenter Orthopaedic Outcomes Network (MOON) Shoulder Instability Consortium and to identify prognostic factors associated with successful return to sport at 2 years postoperatively. Case series; Level of evidence, 4. Overhead athletes undergoing primary arthroscopic anterior shoulder stabilization as part of the MOON Shoulder Instability Consortium were identified for analysis. Primary outcomes included the rate of recurrent instability, defined as any patient reporting recurrent dislocation or reoperation attributed to persistent instability, and return to sport at 2 years postoperatively. Secondary outcomes included the Western Ontario Shoulder Instability Index and Kerlan-Jobe Orthopaedic Clinic Shoulder and Elbow questionnaire score. Univariate regression analysis was performed to identify patient and surgical factors predictive of return to sport at short-term follow-up. A total of 49 athletes were identified for inclusion. At 2-year follow-up, 31 (63%) athletes reported returning to sport. Of those returning to sport, 22 athletes (45% of the study population) were able to return to their previous levels of competition (nonrefereed, refereed, or professional) in at least 1 overhead sport. Two patients (4.1%) underwent revision stabilization, although 14 (28.6%) reported subjective apprehension or looseness. Age ( P = .87), sex ( P = .82), and baseline level of competition ( P = .37) were not predictive of return to sport. No difference in range of motion in all planes ( P > .05) and Western Ontario Shoulder Instability Index scores (78.0 vs 80.1, P = .73) was noted between those who reported returning to sport and those who did not. Primary arthroscopic anterior shoulder stabilization in overhead athletes is associated with a low rate of recurrent stabilization surgery. Return to overhead athletics at short-term follow-up is lower than that previously reported for the general athletic population.


Non-conformal coarse-grained potentials for water.
Water is a notoriously difficult substance to model both accurately and efficiently. Here, we focus on descriptions with a single coarse-grained particle per molecule using the so-called approximate non-conformal and generalized Stockmayer potentials as the starting points. They are fitted using the radial distribution function and the liquid-gas density profile of the atomistic extended simple point charge (SPC/E) model by downhill simplex optimization. We compare the results with monatomic water (mW), ELBA, and direct iterative Boltzmann inversion of SPC/E. The results show that symmetrical potentials result in non-transferable models, that is, they need to be reparametrized for new state points. This indicates that transferability may require more complex models. Furthermore, the results also show that the addition of a point dipole is not sufficient to make the potentials accurate and transferable to different temperatures (300 K-500 K) and pressures without an appropriate choice of properties as targets during model optimization.


Modified Bovine Milk Exosomes for Doxorubicin Delivery to Triple-Negative Breast Cancer Cells.
Biological nanoparticles, such as exosomes, offer an approach to drug delivery because of their innate ability to transport biomolecules. Exosomes are derived from cells and an integral component of cellular communication. However, the cellular cargo of human exosomes could negatively impact their use as a safe drug carrier. Additionally, exosomes have the intrinsic yet enigmatic, targeting characteristics of complex cellular communication. Hence, harnessing the natural transport abilities of exosomes for drug delivery requires predictably targeting these biological nanoparticles. This manuscript describes the use of two chemical modifications, incorporating a neuropilin receptor agonist peptide (iRGD) and a hypoxia-responsive lipid for targeting and release of an encapsulated drug from bovine milk exosomes to triple-negative breast cancer cells. Triple-negative breast cancer is a very aggressive and deadly form of malignancy with limited treatment options. Incorporation of both the iRGD peptide and hypoxia-responsive lipid into the lipid bilayer of bovine milk exosomes and encapsulation of the anticancer drug, doxorubicin, created the peptide targeted, hypoxia-responsive bovine milk exosomes, iDHRX. Initial studies confirmed the presence of iRGD peptide and the exosomes' ability to target the αvβ3 integrin, overexpressed on triple-negative breast cancer cells' surface. These modified exosomes were stable under normoxic conditions but fragmented in the reducing microenvironment created by 10 mM glutathione. In vitro cellular internalization studies in monolayer and three-dimensional (3D) spheroids of triple-negative breast cancer cells confirmed the cell-killing ability of iDHRX. Cell viability of 50% was reached at 10 μM iDHRX in the 3D spheroid models using four different triple-negative breast cancer cell lines. Overall, the tumor penetrating, hypoxia-responsive exosomes encapsulating doxorubicin would be effective in reducing triple-negative breast cancer cells' survival.


[Changes in calcium signaling in mammalian nociceptive neurons in diabetes mellitus].
The changes in neuronal Ca2+ homeostasis were studied on dorsal horn neurons from spinal cord rat slices and freshly isolated dorsal root ganglion neurons of mice in control condition and under streptozotocin (STZ)-induced diabetes. The cytoplasmic free Ca2+ concentration ([Ca2+]i) was measured using fura-2 and indo-1 based microfluorimetry. The recovery of depolarization-induced [Ca2+]i increase was delayed in diabetic neurons compared with normal animals. The amplitude of calcium release from caffeine-sensitive endoplasmic reticulum calcium stores became significantly smaller in diabetic neurons. The participation of mitochondria in [Ca2+]i homeostasis was determined by investigation of changes which occurred after addition of mitochondrial protonophore (CCCP) to the extracellular solution. In control cells 10 (M CCCP applied before membrane depolarization induced an increase of the amplitude of depolarization-induced [Ca2+]i transients and disappearance of their delayed recovery, indicating the participation of mitochondria in fast uptake of Ca2+ ions from the cytosol during the peak of the transient and subsequent slow release them back during its decay. In neurons from diabetic animals the increase of the peak transient amplitude under the action of CCCP became diminished, and the delayed elevation of [Ca2+]i disappeared in small size neurons. We conclude that streptozotocin-induced diabetes is associated with prominent changes in the mechanisms responsible for [Ca2+]i regulation in neurones of the nociceptive system, which presumably include a slow down of Ca2+ elimination from the cytoplasm by endoplasmic reticulum and mitochondria.


Role of osmotic and hydrostatic pressures in bacteriophage genome ejection.
A critical step in the bacteriophage life cycle is genome ejection into host bacteria. The ejection process for double-stranded DNA phages has been studied thoroughly in vitro, where after triggering with the cellular receptor the genome ejects into a buffer. The experimental data have been interpreted in terms of the decrease in free energy of the densely packed DNA associated with genome ejection. Here we detail a simple model of genome ejection in terms of the hydrostatic and osmotic pressures inside the phage, a bacterium, and a buffer solution or culture medium. We argue that the hydrodynamic flow associated with the water movement from the buffer solution into the phage capsid and further drainage into the bacterial cytoplasm, driven by the osmotic gradient between the bacterial cytoplasm and culture medium, provides an alternative mechanism for phage genome ejection in vivo; the mechanism is perfectly consistent with phage genome ejection in vitro.


Conservative treatment of acute appendicitis.
Acute appendicitis has been considered by surgeons a progressive disease leading to perforation for more than 100 years. In the last decades the theories about this concept gained attention, especially in adults. However, appendectomy for acute appendicitis remains the most common urgent/emergent surgical procedure. At present, accumulating evidences are showing the changing in clinical practice towards the non-operative management of several cases of acute appendicitis either non-complicated or complicated. The present review aims to show the literature results regarding the non-operative management of acute appendicitis in non-complicated and in complicated cases.


Nonmyeloablative stem cell transplant in a patient with advanced systemic sclerosis and systemic lupus erythematosus.
Systemic sclerosis (SSc) is an uncommon connective tissue disease characterized by excessive collagen deposition within the skin and internal organs. Most patients with diffuse severe SSc are treated with immunosuppressive agents, but patients with advanced disease have very high 5-year mortality rates despite adequate therapy. We describe a patient with both diffuse cutaneous SSc and systemic lupus erythematosus who showed mixed chimerism 29 months after undergoing nonmyeloablative stem cell transplant. She experienced remission of both diseases.


Mitochondria as Key Players in the Pathogenesis and Treatment of Rheumatoid Arthritis.
Mitochondria are major energy-producing organelles that have central roles in cellular metabolism. They also act as important signalling hubs, and their dynamic regulation in response to stress signals helps to dictate the stress response of the cell. Rheumatoid arthritis is an inflammatory and autoimmune disease with high prevalence and complex aetiology. Mitochondrial activity affects differentiation, activation and survival of immune and non-immune cells that contribute to the pathogenesis of this disease. This review outlines what is known about the role of mitochondria in rheumatoid arthritis pathogenesis, and how current and future therapeutic strategies can function through modulation of mitochondrial activity. We also highlight areas of this topic that warrant further study. As producers of energy and of metabolites such as succinate and citrate, mitochondria help to shape the inflammatory phenotype of leukocytes during disease. Mitochondrial components can directly stimulate immune receptors by acting as damage-associated molecular patterns, which could represent an initiating factor for the development of sterile inflammation. Mitochondria are also an important source of intracellular reactive oxygen species, and facilitate the activation of the NLRP3 inflammasome, which produces cytokines linked to disease symptoms in rheumatoid arthritis. The fact that mitochondria contain their own genetic material renders them susceptible to mutation, which can propagate their dysfunction and immunostimulatory potential. Several drugs currently used for the treatment of rheumatoid arthritis regulate mitochondrial function either directly or indirectly. These actions contribute to their immunomodulatory functions, but can also lead to adverse effects. Metabolic and mitochondrial pathways are attractive targets for future anti-rheumatic drugs, however many questions still remain about the precise role of mitochondrial activity in different cell types in rheumatoid arthritis.


Evidences of increasing risk of dirofilarioses in southern Italy.
Given the spread of Aedes albopictus from northern to southern Italy, and the lack of updated data on Dirofilaria infections, this study was carried out to assess the infection risk for dogs and cats in Apulia region. During a 2-year study, 175 A. albopictus female specimens and samples of blood from 427 dogs (309 privately owned dogs and 118 shelter dogs) and 12 cats were collected. All blood samples were subjected to a modified Knott method, to a test for the detection of circulating Dirofilaria immitis antigen, and to a Dirofilaria species-specific real-time PCR for the simultaneous detection of D. immitis and Dirofilaria repens, targeting on partial cytochrome oxidase subunit 1 and internal transcribed spacer-2, respectively. Two abdomen and one thorax pools from A. albopictus were positive for D. immitis, with minimum infection rates of 1.14 and 0.51, respectively, and a probability of a single positive specimen to be infected of P = 0.6 % (95 % confidence interval (CI) = 0.12-1.73). Out of 439 examined subjects, 22 (5.0 %) tested positive for Dirofilaria spp. in at least one diagnostic test. A specific D. immitis infestation rate of 3.5 % was found among the privately owned dogs, while shelter dogs tested positive only for D. repens with a prevalence of 3.4 %; one cat tested molecularly positive for D. immitis. There was a significantly higher rate of positivity among guard dogs for D. immitis (odds ratio, 6.24, 95 % CI, 1.26-25.28; P < 0.05). The increasing risk of D. immitis infection in southern Italy is supported by the noteworthy positivity of A. albopictus populations and the cat. Our data highlight the usefulness to include filarioid infestation in routine diagnosis.


Human immunodeficiency virus type 1 Vif protein binds to the Pr55Gag precursor.
The Vif protein of human immunodeficiency virus type 1 is required for productive replication in peripheral blood lymphocytes. Previous reports suggest that vif-deleted viruses are limited in replication because of a defect in the late steps of the virus life cycle. One of the remaining questions is to determine whether the functional role of Vif involves a specific interaction with virus core proteins. In this study, we demonstrate a direct interaction between Vif and the Pr55Gag precursor in vitro as well as in infected cells. No interaction is observed between Vif and the mature capsid protein. The Pr55Gag-Vif interaction is detected (i) in the glutathione S-transferase system, with in vitro-translated proteins demonstrating a critical role of the NC p7 domain of the Gag precursor; (ii) with proteins expressed in infected cells; and (iii) by coimmunoprecipitation experiments. Deletion of the C-terminal 22 amino acids of Vif abolishes its interaction with the Pr55Gag precursor. Furthermore, point mutations in the C-terminal domain of Vif which have been previously shown to abolish virus infectivity and binding to cell membranes dramatically decrease the Gag-Vif interaction. These results suggest that the interaction between Vif and the pr55Gag precursor is a critical determinant of Vif function.


Iohexol for cervical myelography via C1-C2 puncture: study of efficacy and adverse reactions.
Cervical myelography with iohexol via C1-C2 puncture was performed in 30 patients in two medical centers using a concentration of 240 mg l/ml. The study demonstrated iohexol to be a safe contrast medium without significant changes in neurologic and physical examination, vital signs, electrocardiogram, or hematologic or blood chemistry parameters. Fifteen patients had electroencephalograms (EEGs); two were abnormal. In one patient the baseline EEG demonstrated nonspecific slow waves in the temporal regions bilaterally that remained unchanged after myelography. In the second patient, transient changes in the left hemisphere during either hyperventilation or photic stimulation on postmyelographic EEG had not been present on the baseline recording. The relation of these changes to the drug remains unclear. Iohexol was found to be an efficacious myelographic contrast agent, with good to excellent myelograms in 93% of cases. Headache occurred in 13% and nausea in 3%.


The BCR gene encodes a novel serine/threonine kinase activity within a single exon.
Sequences encoded by the first exon of BCR that bind to the ABL SH2 domain are essential for the activation of the ABL tyrosine kinase and transforming potential of the chimeric BCR-ABL oncogene. The normal cellular BCR gene encodes a 160,000 dalton phosphoprotein associated with a serine/threonine kinase activity, but it shows only weak dispersed homologies to protein kinases. p160c-BCR was purified to apparent homogeneity as an oligomer of greater than 600,000 daltons that contains autophosphorylation activity and transphosphorylation activity for several protein substrates. A region containing paired cysteine residues within the 426 amino acids encoded by the first exon of BCR is essential for its novel phosphotransferase activity, which overlaps with the strong SH2-binding regions. The recent demonstration of a GTPase-activating function within the C-terminal portion of BCR suggests that the protein kinase and SH2-binding domains may work in concert with other regions of the molecule in intracellular signalling processes.


Optical Sorter-Based Selection Effectively Identifies Soft Red Winter Wheat Breeding Lines With Fhb1 and Enhances FHB Resistance in Lines With and Without Fhb1.
Previous results from our lab have shown that using an optical sorter to identify Fusarium head blight (FHB) resistant breeding lines was effective at reducing the toxin deoxynivalenol (DON) and FHB-associated kernel damage. In this paper we quantified the proportion of desirable genotypes at FHB resistance QTL in lines from three selection cycles of optical sorting. Breeding lines were genotyped at loci on chromosomes 3BS, 2DL, and 5A using the following DNA markers: TaHRC, CFD233, and GWM304. TaHRC is a KASP marker for Fhb1, a major FHB resistance QTL on chromosome 3BS. CFD233 is an SSR marker for Qfhs.nau-2DL on chromosome 2DL. GWM304 is an SSR marker for Qfhs.ifa-5A on chromosome 5A. Sorter selection was effective at identifying lines that had the resistant genotype at TaHRC; in other words, the sorter was able to identify lines with resistance alleles at Fhb1. The sorter was less effective at selecting for the resistant genotype at CFD233 and GWM304. However, the proportion of lines with resistant genotypes at GWM304 did increase with additional sorter selection, just not to the degree that was observed for the Fhb1-associated marker. The proportion of lines with resistant alleles at CFD233 did not show a consistent trend. In addition to increasing the proportion of lines with Fhb1 and Qfhs.ifa-5A each selection cycle, optical sorter-based mass selection enhanced FHB resistance in different marker genotype combinations evaluated in this study. For example, there were net reductions in DON and kernel damage after two cycles of sorter selection in 15X110601S07002, a line with Fhb1, with Qfhs.nau-2DL, and with Qfhs.ifa-5A; final C3 DON levels were 63% of the resistant check (KY02C-3005-25). Kernel damage was also reduced in 15X110601A08221 a line without Fhb1, without Qfhs.nau-2DL, and without Qfhs.ifa-5A. Our findings suggest the increased resistance observed in different marker genotype combinations was conferred by QTL other than Fhb1, QFhs.nau-2DL, and Qfhs.ifa-5, and validate our previous results that the optical sorter is effective at selecting FHB-resistant breeding material.


CENP-A and CENP-B collaborate to create an open centromeric chromatin state.
Centromeres are epigenetically defined via the presence of the histone H3 variant CENP-A. Contacting CENP-A nucleosomes, the constitutive centromere associated network (CCAN) and the kinetochore assemble, connecting the centromere to spindle microtubules during cell division. The DNA-binding centromeric protein CENP-B is involved in maintaining centromere stability and, together with CENP-A, shapes the centromeric chromatin state. The nanoscale organization of centromeric chromatin is not well understood. Here, we use single-molecule fluorescence and cryoelectron microscopy (cryoEM) to show that CENP-A incorporation establishes a dynamic and open chromatin state. The increased dynamics of CENP-A chromatin create an opening for CENP-B DNA access. In turn, bound CENP-B further opens the chromatin fiber structure and induces nucleosomal DNA unwrapping. Finally, removal of CENP-A increases CENP-B mobility in cells. Together, our studies show that the two centromere-specific proteins collaborate to reshape chromatin structure, enabling the binding of centromeric factors and establishing a centromeric chromatin state.


External cardioversion of atrial fibrillation: role of paddle position on technical efficacy and energy requirements.
To define the effect of defibrillator paddle position on technical success and dc shock energy requirements of external cardioversion of atrial fibrillation. 301 patients (mean (SD) age 62 (11) years) with stable atrial fibrillation were randomly assigned to elective external cardioversion using anterolateral paddle position (ventricular apex-right infraclavicular area; group AL (151 patients)) or anteroposterior paddle position (sternal body-angle of the left scapula; group AP (150 patients)). A step up protocol was used, delivering a 3 J/kg body weight dc shock, then a 4 J/kg shock (maximum 360 J), and finally a second 4 J/kg shock using the alternative paddle location. The two groups were comparable for the all clinical variables evaluated. The cumulative percentage of patients successfully converted to sinus rhythm was 58% in group AL and 67% in group AP with low energy dc shock (NS); this rose to 76% in group AL and to 87% in group AP with high energy dc shock (p = 0.013). Thirty seven patients in group AL and 19 in group AP experienced dc shock with the alternative paddle position; atrial fibrillation persisted in 10/37 in group AL and in 10/19 in group AP. Mean dc shock energy requirements were lower for group AP patients than for group AL patients, at 383 (235) v 451 (287) J, p = 0.025. Arrhythmia duration was the only factor that affected the technical success of external cardioversion (successful: 281 patients, 80 (109) days; unsuccessful: 20 patients, 193 (229) days; p < 0.0001). The success rate was lower if atrial fibrillation persisted for > 6 months: 29 of 37 (78%) v 252 of 264 (95%); p = 0.0001. An anteroposterior defibrillator paddle position is superior to an anterolateral location with regard to technical success in external cardioversion of stable atrial fibrillation, and permits lower dc shock energy requirements. Arrhythmia duration is the only clinical variable that can limit the restoration of sinus rhythm.


Determination of safety factor for defibrillator waveforms in cultured heart cells.
We studied excitation thresholds and arrhythmias produced in cultured chick embryo myocardial cells subjected to electric shocks using rectangular, untruncated resistor-capacitor (RC), and critically damped resistor-inductor-capacitor (RLC) waveforms with variable intensities while photocell mechanograms were recorded. Strength-duration curves for excitation and production of a specific postshock arrhythmia (4-s arrest) were constructed. Excitation curves closely resembled those for in situ defibrillation threshold (or specific % success). The ratio between the shock intensity producing a 4-s arrest and that producing excitation at each duration (termed the "safety factor") was determined. Waveforms with a large safety factor in vitro defibrillated most effectively in situ with little postshock dysfunction. Waveforms with low safety factors had a low rate of success in situ and produced much postshock dysfunction. Safety factor of monophasic clinical waveforms were lower than that of the 5-ms rectangular wave. The close correspondence between in vitro safety factor and in situ defibrillating effectiveness, as reported in the literature, suggests that the cell culture system is an effective screening system for determining waveforms that will improve the efficacy and safety of defibrillation procedures.


Production of the neurotoxin BMAA by a marine cyanobacterium.
Diverse species of cyanobacteria have recently been discovered to produce the neurotoxic non-protein amino acid beta-methylamino-L-alanine (BMAA). In Guam, BMAA has been studied as a possible environmental toxin in the diets of indigenous Chamorro people known to have high levels of Amyotrophic Lateral Sclerosis/ Parkinsonism Dementia Complex (ALS/PDC). BMAA has been found to accumulate in brain tissues of patients with progressive neurodegenerative illness in North America. In Guam, BMAA was found to be produced by endosymbiotic cyanobacteria of the genus Nostoc which live in specialized cycad roots. We here report detection of BMAA in laboratory cultures of a free-living marine species of Nostoc. We successfully detected BMAA in this marine species of Nostoc with five different methods: HPLC-FD, UPLC-UV, Amino Acid Analyzer, LC/MS, and Triple Quadrupole LC/MS/MS. This consensus of five different analytical methods unequivocally demonstrates the presence of BMAA in this marine cyanobacterium. Since protein-associated BMAA can accumulate in increasing levels within food chains, it is possible that biomagnification of BMAA could occur in marine ecosystems similar to the biomagnification of BMAA in terrestrial ecosystems. Production of BMAA by marine cyanobacteria may represent another route of human exposure to BMAA. Since BMAA at low concentrations causes the death of motor neurons, low levels of BMAA exposure may trigger motor neuron disease in genetically vulnerable individuals.


A statistical framework for differential pseudotime analysis with multiple single-cell RNA-seq samples.
Pseudotime analysis with single-cell RNA-sequencing (scRNA-seq) data has been widely used to study dynamic gene regulatory programs along continuous biological processes. While many methods have been developed to infer the pseudotemporal trajectories of cells within a biological sample, it remains a challenge to compare pseudotemporal patterns with multiple samples (or replicates) across different experimental conditions. Here, we introduce Lamian, a comprehensive and statistically-rigorous computational framework for differential multi-sample pseudotime analysis. Lamian can be used to identify changes in a biological process associated with sample covariates, such as different biological conditions while adjusting for batch effects, and to detect changes in gene expression, cell density, and topology of a pseudotemporal trajectory. Unlike existing methods that ignore sample variability, Lamian draws statistical inference after accounting for cross-sample variability and hence substantially reduces sample-specific false discoveries that are not generalizable to new samples. Using both real scRNA-seq and simulation data, including an analysis of differential immune response programs between COVID-19 patients with different disease severity levels, we demonstrate the advantages of Lamian in decoding cellular gene expression programs in continuous biological processes.


Clinically meaningful interpretation of pediatric health-related quality of life in sickle cell disease.
Health-related quality of life (HRQL) measures provide information about disease assessment; however, health care providers may be reluctant to use HRQL assessments as scores can be difficult to interpret. We sought to identify levels for impaired pain-related HRQL in children with sickle cell disease (SCD). Children (n=251) completed the Pediatric Quality of Life Inventory (PedsQL) Generic Core Scales and PedsQL SCD Module in a multisite study. Using children's item scores on the Pain and Hurt and Pain Impact scales of the PedsQL SCD Module, High, Intermediate, and Low Functioning groups were created. We compared functioning groups with the Pain and Hurt and Pain Impact scale scores to determine levels representing high and low HRQL. These scores were compared with disease severity and the PedsQL Generic Core Scales. Scores of ≤60 on the PedsQL SCD Pain and Hurt and Pain Impact scales were associated with severe disease and met requirements for impaired functioning on the PedsQL Generic Core Scales. Scores of ≥81 on the Pain and Hurt and the Pain Impact scales can be considered consistent with good HRQL in those domains in SCD. Alternately, scores of ≤60 are cause for concern and suggest areas of HRQL impairment in SCD.


Influence of intrauterine dispositive in human papillomavirus clearance.
An important inverse relation between IUD use and risk of cervical cancer has been proved. Women who used IUD had half the risk of developing cervical cancer. The mechanism how IUD is a protector factor is still unknown. Could be improving the clearance of HPV infection or stopping progression to cancer from preneoplasic lesion. The aim of the study is to check if IUD increases HPV clearance, that is, checking if after 1 year HPV infection disappears in more patients using IUD than those not using it. This is a cohort case-control prospective study, carried out in Universitary Hospital La Zarzuela in Madrid, Spain, performed between October 2015 and April 2018. No pregnant women between 25-50 years old, with HPV cervical infection were enrolled. We separated the participants into two groups: an IUD group, with women starting using IUD and non IUD group, with women using any other contraceptive method or none. HPV genotyping of cervical cytology samples were performed initially on enrolment day and one year after. 254 participants were enrolled at the beginning of the study, 85 in the IUD group and 169 in the control group (non IUD). 179 participants completed the study, 54 (31%) in IUD group and 120 participants in non IUD group (69%). 38 women from IUD group cleared HPV infection (69.5%) and 65 women from control group cleared it (54.2%) (p = 0.044). An association in logistic regression was observed in HPV clearance with different factors. Firstly, higher percentage of patients with IUD clear the HPV infection than those in control group significantly associated (OR = 0.698, CI 95%; 0.251-0.998, p = 0.046). Clearance was higher in patients with low-risk HPV infection comparing with high risk HPV (OR = 1.078, CI 95%; 1.126-4.6.281, p = 0.026) and in patients with only one HPV type than those with more than one (OR = 0.194, CI 95%; 0.084-0.403, p < 0.001). In Spanish women with HPV infection, the HPV clearance between IUD and non IUD groups show results with statistical significance, patients with IUD have higher clearance rates. There were differences also between suffering one HPV type or more than one, and having low-risk HPV or high-risk HPV infection.


Intrathoracic chordoma presenting as a posterior mediastinal tumor.
Chordomas are tumors originating from remnants of the notochord, most commonly found in the cranial and caudal regions. This is a report of the seventh intrathoracic chordoma in the world literature and the second to be reported in a woman. Histologically the physalipherous (blister-bearing) cell is characteristic of the tumor. Complete surgical excision is rarely successful and recurrence with or without metastasis is the rule.


Combining Awake Anesthesia with Minimal Invasive Surgery Optimizes Intraoperative Surgical Spinal Cord Stimulation Lead Placement.
Spinal cord stimulation (SCS) is an effective and validated treatment to address chronic refractory neuropathic pain in persistent spinal pain syndrome-type 2 (PSPS-T2) patients. Surgical SCS lead placement is traditionally performed under general anesthesia due to its invasiveness. In parallel, recent works have suggested that awake anesthesia (AA), consisting of target controlled intra-venous anesthesia (TCIVA), could be an interesting tool to optimize lead anatomical placement using patient intra-operative feedback. We hypothesized that combining AA with minimal invasive surgery (MIS) could improve SCS outcomes. The goal of this study was to evaluate SCS lead performance (defined by the area of pain adequately covered by paraesthesia generated via SCS), using an intraoperative objective quantitative mapping tool, and secondarily, to assess pain relief, functional improvement and change in quality of life with a composite score. We analyzed data from a prospective multicenter study (ESTIMET) to compare the outcomes of 115 patients implanted with MIS under AA (MISAA group) or general anesthesia (MISGA group), or by laminectomy under general anesthesia (LGA group). All in all, awake surgery appears to show significantly better performance than general anesthesia in terms of patient pain coverage (65% vs. 34-62%), pain surface (50-76% vs. 50-61%) and pain intensity (65% vs. 35-40%), as well as improved secondary outcomes (quality of life, functional disability and depression). One step further, our results suggest that MISAA combined with intra-operative hypnosis could potentialize patient intraoperative cooperation and could be proposed as a personalized package offered to PSPS-T2 patients eligible for SCS implantation in highly dedicated neuromodulation centers.


[Transient global amnesia: study of 26 cases].
The clinical picture and investigation of 26 patients (16 males and 10 females) with diagnosis of transient global amnesia (TGA) are reported. Age ranged from 51 to 78 years at the time of TGA, which occurred mor often between 60 and 70 year-old people. Three patients presented more than one episode (3, 4 and 5). Precipitating factors were identified in 8 cases (emotional stress in 7 and physical exercise in 1). Risk factors for cerebrovascular disease were found in 13 cases, mainly hypertension (9 cases) and diabetes (3 cases). EEG was normal in 20 cases and disclosed diffuse delta waves in 2, temporal delta waves in 1 and temporal theta waves in another patient. CAT scan showed no abnormalities in 3 cases and ischemia in the vertebro-basilar system in another 2. Brain angiography was normal in 1 case and showed abnormalities in the vertebro-basilar system arteries in 3. During the follow-up period, which ranged from 1 to 84 months, no neurologic deterioration was seen. The role of risk factors for vascular diseases, epilepsy and migraine in the development of TGA is discussed.


The erythemal ultraviolet exposure for humans in greenhouses.
A spectrum evaluator has been employed to evaluate the erythemal exposure in late spring and late summer at three sites and five orientations at each site inside a glass greenhouse with black shadecloth over the glass roof. The maximum in the erythemal irradiance in the greenhouse is not necessarily at noon. Over a day, the maximum erythemal exposure occurred on the eastern side of the greenhouse in the morning and on the western side in the afternoon. The erythemal irradiance on the eastern side in the morning was higher by 26% and 50% for horizontal and vertical surfaces respectively compared with the same site at noon. On the western side the irradiance was higher by 45% and 78% for the horizontal and vertical surfaces respectively compared with the same site at noon. The erythemal irradiance inside the greenhouse does not vary as much during the day as it does outside; for example, for horizontal surfaces, the ratio of the erythemal irradiance outside to the average inside the greenhouse varies from 66 to 112 to 74 for the morning, noon and afternoon periods respectively. Over a 6 h period, the erythemal exposure to the shoulder for a standing posture in the greenhouse was 5 mJ cm-2.


Development of a classical swine fever subunit marker vaccine and companion diagnostic test.
The development of a classical swine fever (CSF) subunit marker vaccine, based on viral envelope glycoprotein E2, and a companion diagnostic test, based on a second viral envelope glycoprotein E(RNS), will be described. Important properties of the vaccine, such as onset and duration of immunity, and prevention of horizontal and vertical transmission of virus were evaluated. A single dose of the vaccine protected pigs against clinical signs of CSF, following intranasal challenge with 100LD(50) of virulent classical swine fever virus (CSFV) at 2 weeks after vaccination. However, challenge virus transmission to unvaccinated sentinels was not always completely inhibited at this time point. From 3 weeks up to 6 months after vaccination, pigs were protected against clinical signs of CSF, and no longer transmitted challenge virus to unvaccinated sentinels. In contrast, unvaccinated control pigs died within 2 weeks after challenge. We also evaluated transmission of challenge virus in a setup enabling determination of the reproduction ratio (R value) of the virus. In such an experiment, transmission of challenge virus is determined in a fully vaccinated population at different time points after vaccination. Pigs challenged at 1 week after immunization died of CSF, whereas the vaccinated sentinels became infected, seroconverted for E(RNS) antibodies, but survived. At 2 weeks after vaccination, the challenged pigs seroconverted for E(RNS) antibodies, but none of the vaccinated sentinels did. Thus, at 1 week after vaccination, R1, and at 2 weeks, R=0, implying no control or control of an outbreak, respectively. Vertical transmission of CSFV to the immune-incompetent fetus may lead to the birth of highly viraemic, persistently infected piglets which are one of the major sources of virus spread. Protection against transplacental transmission of CSFV in vaccinated sows was, therefore, tested in once and twice vaccinated sows. Only one out of nine once-vaccinated sows transmitted challenge virus to the fetus, whereas none of the nine twice-vaccinated sows did. Finally, our data show that the E(RNS) test detects CSFV-specific antibodies in vaccinated or unvaccinated pigs as early as 14 days after infection with a virulent CSF strain. This indicates that the E2 vaccine and companion test fully comply with the marker vaccine concept. This concept implies the possibility of detecting infected animals within a vaccinated population.


Application of the Kohonen artificial neural network in the identification of proteinaceous binders in samples of panel painting using gas chromatography-mass spectrometry.
Historically, three types of proteinaceous matter--casein, egg and animal glue--were used as binders for pigments or as adhesives in easel and wall painting. The relative percentage content of alanine, glycine, valine, leucine, isoleucine, serine, tyrosine, phenylalanine, aspartic acid, glutamic acid, lysine, methionine, proline and hydroxyproline, as determined by GC-MS, is used for binder identification. In this paper we analyse the viability of a multivariate modelling using Kohonen's neural network to characterise the wood adhesive in 16 old samples from Italian panel paintings of the 12-16th centuries. As a training set we use the amino acid composition of 141 samples contributed by the Opificio delle Pietre Dure of Florence (Cultural Heritage Ministry, Italy). Of the 141 samples, 113 were used to train the Kohonen neural network and the remaining 28 as the evaluation set. A specificity and sensitivity of 100% was achieved in training and 92-100% in prediction depending on the assignation criteria employed. The neural network thus trained and evaluated was applied to the old samples, achieving identification of all of them. In addition, the map obtained for each amino acid provides relevant information as to its importance in the characterisation of the sample.


The influence of medical student gender and drug use on the detection of addiction in patients.
Little is known about medical students' interest in their training on drug addiction, their personal experience of consumption, and whether these aspects influence the detection of addiction in patients. Eighty-eight and one half percent considered that drug dependence issues are important to their professional future. The students report consuming alcohol (69%), cigarettes (19.5%), and illegal drugs (15.8%). Female students consumed fewer illegal drugs than the men (p =.022). Male students consumed more illegal drugs more frequently (p =.005), knew more consumers (p =.023), and those who drink alcohol consumed more illegal drugs than women who drink alcohol (p <.005). Drug and alcohol consumption among medical students may serve to normalize consumption and thus, may prevent the detection of addicts. It is important to educate and raise awareness about drugs and alcohol use, as this may influence detection. The focus should be particularly on the male group.


Does stage at diagnosis influence the observed relationship between socioeconomic status and breast cancer incidence, case-fatality, and mortality?
Historically, lower socioeconomic status (SES) has been reported to be associated with decreased breast cancer incidence and mortality and increased case-fatality, although recent trends in breast cancer screening and treatment may alter these relationships. This study assessed the associations between SES and breast cancer incidence, case-fatality, and mortality by stage of disease at diagnosis using recent data in the United States. Breast cancer incidence and survival data from the Surveillance, Epidemiology, and End Results (SEER) tumor registry for black and white women aged 55 and above were linked to county level SES and population data based on place of residence. Poisson regression was used to calculate age-adjusted relative rates associated with SES levels and breast cancer incidence, case-fatality, and mortality. As SES decreased, localized breast cancer incidence rates decreased, while incidence rates of distant disease increased. Five-year localized and regional breast cancer case-fatality rates increased as SES decreased. Localized breast cancer mortality rates decreased as SES declined, whereas regional breast cancer mortality rates tended to increase. These results confirm some previously reported findings and suggest that associations between lower SES and lower localized breast cancer mortality rates are influenced mainly by underlying associations between SES and localized breast cancer incidence, whereas regional breast cancer mortality rates appear to reflect the underlying association between SES and regional case-fatality rates.


News on tobacco and public attitudes toward smokefree air policies in the United States.
Smoking bans are a central component of comprehensive tobacco control programs, and an increasing number of states and localities have adopted them. Public support is critical as efforts continue to extend protection from secondhand smoke to all. We examine the relationship between state print news coverage of tobacco and public sentiment towards smoking bans. We utilize two data sources: the February 2002 Current Population Survey Tobacco Use Supplement, and articles from 2 months in a database comprised of content-analyzed newspaper articles on tobacco from all daily U.S. newspapers. We control for demographic and state-level tobacco covariates, and use logistic regression to identify associations between news coverage and attitudes towards policy initiatives by smoking status. We find no association between tone of news coverage and any outcome measure and little evidence of a relationship between the volume of overall tobacco news and attitudes towards smoking bans. Our data suggest that people exposed to more news on secondhand smoke issues are less likely to support restaurant smoking bans. Higher volume of secondhand smoke news is also predictive of lower support for bar bans among recent quitters. Support for hospital bans is, however, largely independent of news coverage. We suggest two alternate explanations for our findings. First, for certain people, greater news volume on secondhand smoke may lead to lower support for bans. Alternatively, secondhand smoke may be more newsworthy in states where bans are more controversial.


Venous occlusion with lidocaine for preventing propofol induced pain. A prospective double-blind randomized study.
Pain is a well-known complication of intravenous administration of propofol, and to find out the optimal method to decrease this pain, we studied 4 methods of delivering propofol. The study took place at Jordan University Hospital, Amman, Jordan between November 2004 and March 2005 on 200 patients. The patients were divided into 4 groups, group I (n=50), the control group, propofol 1% was given alone. Group II (n=50), patients received propofol 1% premixed with 40 mg of lidocaine. Group III (n=50), patients received propofol 1% 60 seconds after giving 40 mg of lidocaine. Group IV (n=50), patients had venous occlusion for 60 seconds with the use of lidocaine 1% (40 mg), followed by release of the occlusion and administration of the propofol. Pain was assessed during injection and categorized into: no pain, pain, and pain with behavioral changes. In group I (control), 35 patient complained of pain, compared to 26 in group II, 23 in group III, and 7 patients in group IV, with a significant reduction in the incidence and intensity of pain in group II, III, and IV compared with the control (p<0.005). The best reduction of intensity and incidence was achieved in group VI, when compared with groups I, II and III (p<0.005), with no statistical difference between group II and III when compared with each other. Of the 4 methods studied, the optimal method to decrease the incidence and intensity of pain resulting from propofol injection is to inject lidocaine while applying venous occlusion for 60 seconds prior to administering propofol.


Modulation of collagenase 3 in human osteoarthritic cartilage by activation of extracellular transforming growth factor beta: role of furin convertase.
Treatment of normal cartilage with transforming growth factor beta (TGFbeta) can increase the synthesis of collagenase 3 by chondrocytes and mimic the in situ distribution of this enzyme in osteoarthritic (OA) cartilage, which occurs predominantly in the deep zone. In this study, we examined the elements of the TGFbeta system that are potentially relevant to this effect. TGFbeta1 and TGFbeta2 levels in cultured cartilage explants were determined by enzyme-linked immunosorbent assay (ELISA). OA cartilage explants were treated with small latent TGFbeta1 complex in the presence of various inhibitors, and collagenase 3 levels were determined by ELISA. The inhibitors were against serine proteases, plasmin, cathepsins, furin, and a neutralizing antibody against the mannose-6 phosphate/ insulin-like growth factor 2 receptor (M6P/IGF-2R). Small latent TGFbeta1, TGFbeta receptor types I, II, and III (TGFbetaRI, RII, and RIII), M6P/IGF-2R, and furin were immunolocalized in cartilage. Our data showed that latent TGFbeta1 is the major isoform that is synthesized; levels of 17.2 +/-1.7 pg/mg and 1.1 +/- 0.3 pg/mg tissue wet weight (mean +/- SEM) were found for total TGFbeta1 and TGFbeta2, respectively, in OA cartilage. A general serine protease inhibitor abrogated activation of both endogenous and exogenous small latent TGFbeta1. Plasmin and furin inhibitors and anti-M6P/IGF-2R reduced the levels of exogenous small latent TGFbeta1 complex-induced collagenase 3 by 33%, 95%, and 76%, respectively, but the cathepsin inhibitor had no effect. Immunolocalization of the small latent TGFbeta1 complex as well as of TGFbetaRI and RII revealed a statistically significant increase in the chondrocyte score in only the deep zone of OA cartilage. The M6P/IGF-2R level was significantly higher in OA cartilage in both the superficial and deep zones. Furin was found in normal cartilage exclusively in the superficial zone, whereas in OA cartilage, a level similar to that in normal cartilage was found in the superficial zone, but a significantly higher cell score (mean +/- SEM 23.6 +/- 4.7%) was registered in the deep zone. The mechanisms of TGFbeta activation/ activity with regard to collagenase 3 modulation in cartilage appear to be controlled by furin convertase with or without M6P/IGF-2R. These factors and the small latent TGFbeta complex are increased in the deep zone of OA cartilage, corresponding to the preferential site of collagenase 3 production.


Anaesthesia for neonatal surgical emergencies in a semi-urban hospital, Nigeria.
To establish the techniques of anaesthesia for neonatal surgical emergencies in a semi-urban hospital in Nigeria, assess their adequacy and make recommendations to improve our practice. Retrospective study. Obafemi Awolowo University Teaching Hospital, Ile-lfe, Nigeria, from January 1990 to December 2000. One hundred neonates aged one to twenty eighty days. One hundred neonatal surgical emergencies were operated but only 76 case notes were available for review. Three hundred and fifty eight elective neonatal surgeries were done during the same period. The ASA classification were: ASA IE=10, 2E=25, 3E=28, 4E =12 and 5E=1. The mean weight was 2.66+/ 0.52 kg. Over 95% of the cases were done under general anaesthesia. Anorectal malformations and intestinal obstruction were the most common indications for surgery (64.5%). Nurse anaesthetists gave over 50% of the anaesthetics. Peri-operative adverse events such as tachycardia, respiratory distress, aspiration and hypothermia were recorded in 11.8% of the cases. Mortality was 39.2%. General anaesthesia is still the main technique of anaesthesia and mortality following surgery is still high. Efforts should be made to train appropriate personnel to provide improved care and thereby reduce morbidity and mortality.


Circulating SH2B1 is associated with an increased risk of gastric cancer.
Gastric cancer (GC) is one of the most common types of cancer in humans and the second leading cause of cancer-associated mortality worldwide. Identifying novel risk factors will facilitate the development of therapeutic strategies to prevent and treat GC. Increased expression of the Src homology 2 B adaptor protein 1 (SH2B1) may stimulate the malignant progression of lung cancer, esophageal cancer and neuroblastoma. However, its function in GC has not yet been investigated. To identify whether increased serum SH2B1 is a risk factor for GC, the present study performed a nested case-control study of patients within the Chinese cohort study. Levels of serum SH2B1 were measured in 563 patients diagnosed with GC during the follow-up period and in 1,126 matched healthy controls. The results demonstrated that high levels of serum SH2B1 were associated with an increased GC risk (odds ratio, 3.23; 95% confidence interval, 2.45-5.65). When analyses were stratified further by sex, age and smoking, an association between increased levels of SH2B1 and GC was identified in males but not in females. Furthermore, the association between SH2B1 levels and GC was more evident in younger than in older participants, and statistically significant in current smokers but not in nonsmokers. These results were not altered following the exclusion of outliers. Furthermore, it was demonstrated that overexpression of SH2B1 contributes to the malignant transformation of normal gastric epithelial cells. Thus, the present study demonstrated that elevated serum SH2B1 levels may increase the risk of GC.


TRPM4 is highly expressed in human colorectal tumor buds and contributes to proliferation, cell cycle, and invasion of colorectal cancer cells.
Transient receptor potential melastatin-4 channel (TRPM4) dysregulation contributes to heart conditions, immune diseases, and cervical and prostate cancer. Up to now, the involvement of TRPM4 in colorectal cancer (CRC) pathophysiology remains unknown. Here, we investigated tumor tissue microarrays from 379 CRC patients and analyzed TRPM4 protein expression, tumor characteristics, and clinical outcome. High TRPM4 protein expression was associated with unfavorable tumor features characteristic for epithelial-mesenchymal transition and infiltrative growth patterns, that is, a high number of tumor buds and a low percentage in tumor border configuration. Compared to CRC cells representing early cancer stages, TRPM4 protein expression was the highest in cells representing late-stage metastatic cancer. Investigation of CRC cell line HCT116 and five CRISPR/cas9 TRPM4 knockout clones demonstrated that TRPM4 exhibited large Na+ current densities (~ 60 pA/pF). In addition, CRISPR/cas9 TRPM4 knockout clones showed a tendency toward decreased migration and invasion, cell viability, and proliferation and exhibited a shift in cell cycle when compared to HCT116. Stable overexpression of TRPM4 (TRPM4 wild-type) in two CRISPR/cas9 TRPM4 knockout clones rescued the decrease in cell viability and cell cycle shift. Stable overexpression of a nonconducting, dominant-negative TRPM4 mutant (TRPM4 D894A) did not rescue the decrease in viability or cell cycle shift. Taken together, these findings pointed to TRPM4 ion channel conductivity as the underlying mechanism for decreased viability and cell cycle shift in the TRPM4 knockout clones. Together with previous findings, our present data suggest that TRPM4 plays a versatile role in cancer cell proliferation, cell cycle, and invasion.


Impact of compulsory participation of medical students in a multiuser online game to learn radiological anatomy and radiological signs within the virtual world Second Life.
Competitive game-based learning within Second Life enables effective teaching of basic radiological anatomy and radiological signs to medical students, with good acceptance and results when students participate voluntarily, but unknown in a compulsory context. The objectives of this study were to reproduce a competitive online game based on self-guided presentations and multiple-choice tests in a mandatory format, to evaluate its development and student perceptions compared to a voluntary edition in 2015 (N = 90). In 2016 and 2017, respectively, 191 and 182 third-year medical students participated in the game as a mandatory course activity. The mean (±SD) score of the game was 74.7% (±19.5%) in 2015, 71.2% (±21.5%) in 2016, and 67.5% (±21.5%) in 2017 (P < 0.01). Participants valued positively the organization and educational contents but found the virtual world less attractive and the game less interesting than in the voluntary edition. The experience globally was rated with 8.2 (±1.5), 7.8 (±1.5), and 7.1 (±1.7) mean points (±SD) in a ten-point scale, in the 2015, 2016, and 2017 editions, respectively (P < 0.05). Competitive learning games within virtual worlds like Second Life have great learning potential in radiology, but the mean score in the game decreased, acceptance of virtual world technology was lower, and opinion about the game was worse with a compulsory participation, and even worse when dropouts were not allowed. Under the conditions in which this study was conducted, learning games in three-dimensional virtual environments should be voluntary to maintain adequate motivation and engagement of medical students.


Biotransformation and bioactivation reactions - 2018 literature highlights.
In the past three decades, ADME sciences have become an integral component of the drug discovery and development process. At the same time, the field has continued to evolve, thus, requiring ADME scientists to be knowledgeable of and engage with diverse aspects of drug assessment: from pharmacology to toxicology, and from in silico modeling to in vitro models and finally in vivo models. Progress in this field requires deliberate exposure to different aspects of ADME; however, this task can seem daunting in the current age of mass information. We hope this review provides a focused and brief summary of a wide array of critical advances over the past year and explains the relevance of this research ( Table 1 ). We divided the articles into categories of (1) drug optimization, (2) metabolites and drug metabolizing enzymes, and (3) bioactivation. This annual review is the fourth of its kind (Baillie et al. 2016 ; Khojasteh et al. 2017 , 2018 ). We have followed the same format we used in previous years in terms of the selection of articles and the authoring of each section. This effort in itself also continues to evolve. I am pleased that Rietjens, Miller, and Mitra have again contributed to this annual review. We would like to welcome Namandjé N. Bumpus, James P. Driscoll, and Donglu Zhang as authors for this year's issue. We strive to maintain a balance of authors from academic and industry settings. We would be pleased to hear your opinions of our commentary, and we extend an invitation to anyone who would like to contribute to a future edition of this review. Cyrus Khojasteh, on behalf of the authors.


Targeting NEDD8-activated cullin-RING ligases for the treatment of cancer.
E3 ubiquitin ligases regulate many dynamic cellular processes important for cancer cell survival. Together with ubiquitin-activating enzyme (E1) and ubiquitin-conjugating enzymes (E2s), E3s catalyze the ubiquitination of numerous protein substrates that are subsequently targeted to the 26S proteasome for degradation. The clinical success of the proteasome inhibitor bortezomib has encouraged the evaluation of other components of the ubiquitin proteasome system for pharmaceutical intervention. Targeting specific E3s is particularly attractive because there is the potential to selectively block the degradation of certain cellular proteins and possibly avoid unwanted effects on other proteins. The cullin-RING ubiquitin E3 ligases (CRLs) represent the largest subfamily of E3s. The requirement that CRLs be activated by NEDD8 modification on the cullin protein offers an "achilles heel" for modulating this entire subfamily. NEDD8-activating enzyme (NAE) catalyzes the first step in the NEDD8 pathway and as such controls the activity of CRLs. In this article, we describe the role of the NEDD8 pathway in activating CRLs and discuss the preclinical findings with a first-in-class NAE inhibitor that is currently in phase I clinical trials for both solid tumor and hematological malignancies. In addition, we speculate where NAE inhibitors may find clinical utility.


[Investigation of antihypoxic properties of short peptides].
The data presented suggest that short regulatory peptides (vilon, epitalon, vesugen and pinealon) have manifested the antihypoxic properties in the model of hypobaric hypoxia. Pinealon (Glu-Asp-Arg) has the most pronounced effect among them. The capability of pinealon to increase the neuronal resistance to hypoxic stress in experiments with prenatal hypoxia has a complex nature. It is based not so much on the inhibition of ROS increase in cells in response to stress as on stimulation of internal antioxidative enzyme system and possibly limiting the excitotoxic effect of N-methyl-D-aspartate.


Using optical coherence tomography and intravascular ultrasound imaging to quantify coronary plaque cap thickness and vulnerability: a pilot study.
Detecting coronary vulnerable plaques in vivo and assessing their vulnerability have been great challenges for clinicians and the research community. Intravascular ultrasound (IVUS) is commonly used in clinical practice for diagnosis and treatment decisions. However, due to IVUS limited resolution (about 150-200 µm), it is not sufficient to detect vulnerable plaques with a threshold cap thickness of 65 µm. Optical Coherence Tomography (OCT) has a resolution of 15-20 µm and can measure fibrous cap thickness more accurately. The aim of this study was to use OCT as the benchmark to obtain patient-specific coronary plaque cap thickness and evaluate the differences between OCT and IVUS fibrous cap quantifications. A cap index with integer values 0-4 was also introduced as a quantitative measure of plaque vulnerability to study plaque vulnerability. Data from 10 patients (mean age: 70.4; m: 6; f: 4) with coronary heart disease who underwent IVUS, OCT, and angiography were collected at Cardiovascular Research Foundation (CRF) using approved protocol with informed consent obtained. 348 slices with lipid core and fibrous caps were selected for study. Convolutional Neural Network (CNN)-based and expert-based data segmentation were performed using established methods previously published. Cap thickness data were extracted to quantify differences between IVUS and OCT measurements. For the 348 slices analyzed, the mean value difference between OCT and IVUS cap thickness measurements was 1.83% (p = 0.031). However, mean value of point-to-point differences was 35.76%. Comparing minimum cap thickness for each plaque, the mean value of the 20 plaque IVUS-OCT differences was 44.46%, ranging from 2.36% to 91.15%. For cap index values assigned to the 348 slices, the disagreement between OCT and IVUS assignments was 25%. However, for the OCT cap index = 2 and 3 groups, the disagreement rates were 91% and 80%, respectively. Furthermore, the observation of cap index changes from baseline to follow-up indicated that IVUS results differed from OCT by 80%. These preliminary results demonstrated that there were significant differences between IVUS and OCT plaque cap thickness measurements. Large-scale patient studies are needed to confirm our findings.


[Acute abdomen of urologic origin].
Presentation of one case of bladder carcinoma which caused acute abdomen following spontaneous perforation. This presentation is extremely rare and should be taken into account when haematuria and abdominal pain appear simultaneously.