Improved overall survival after contralateral risk-reducing mastectomy in BRCA1/2 mutation carriers with a history of unilateral breast cancer: a prospective analysis.
Data on survival of BRCA1/2-associated primary breast cancer (PBC) patients who opt for subsequent contralateral risk-reducing mastectomy (CRRM) are scarce and inconsistent. We examined the efficacy of CRRM on overall survival in mutation carriers with a history of PBC. From a Dutch multicentre cohort, we selected 583 BRCA-associated PBC patients, being diagnosed between 1980 and 2011. Over time, 242 patients (42%) underwent CRRM and 341 patients (58%) remained under surveillance. Survival analyses were performed using Cox models, with CRRM as a time-dependent covariate. The median follow-up after PBC diagnosis was 11.4 years. In the CRRM group, four patients developed contralateral breast cancer (2%), against 64 patients (19%) in the surveillance group (p < 0.001). The mortality was lower in the CRRM group than in the surveillance group (9.6 and 21.6 per 1000 person-years of observation, respectively; adjusted hazard ratio 0.49, 95% confidence interval 0.29-0.82). Survival benefit was especially seen in young PBC patients (<40 years), in patients having a PBC with differentiation grade 1/2 and/or no triple-negative phenotype, and in patients not treated with adjuvant chemotherapy. We conclude that CRRM is associated with improved overall survival in BRCA1/2 mutation carriers with a history of PBC. Further research is warranted to develop a model based on age at diagnosis and tumour and treatment characteristics that can predict survival benefit for specific subgroups of patients, aiming at further personalized counselling and improved decision making.


Insomnia and short-acting benzodiazepine hypnotics.
Insomnia--the chronic inability to obtain the amount and quality of sleep needed for effective daytime function--is a common subjective complaint. Several major causes exist, and many strategems can be used in management. When drugs are indicated, the benzodiazepines are the first choice. Flurazepam, a long-acting compound, has recently been complemented by temazepam, a shorter-acting hypnotic. The pharmacokinetics, actions, and clinical uses of temazepam are reviewed. It is concluded that temazepam is preferable where daytime alertness must be unimpaired, with flurazepam reserved for patients who need daytime sedation.


Structural insight into RNA hairpin folding intermediates.
Hairpins are a ubiquitous secondary structure motif in RNA molecules. Despite their simple structure, there is some debate over whether they fold in a two-state or multi-state manner. We have studied the folding of a small tetraloop hairpin using a serial version of replica exchange molecular dynamics on a distributed computing environment. On the basis of these simulations, we have identified a number of intermediates that are consistent with experimental results. We also find that folding is not simply the reverse of high-temperature unfolding and suggest that this may be a general feature of biomolecular folding.


A 2H solid-state NMR study of lipid clustering by cationic antimicrobial and cell-penetrating peptides in model bacterial membranes.
Domain formation in bacteria-mimetic membranes due to cationic peptide binding was recently proposed based on calorimetric data. We now use (2)H solid-state NMR to critically examine the presence and absence of domains in bacterial membranes containing zwitterionic 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphatidylethanolamine (POPE) and anionic 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphatidylglycerol (POPG) lipids. Chain-perdeuterated POPE and POPG are used in single-component membranes, binary POPE/POPG (3:1) membranes, and membranes containing one of four cationic peptides: two antimicrobial peptides (AMPs) of the β-hairpin family of protegrin-1 (PG-1), and two cell-penetrating peptides (CPPs), HIV TAT and penetratin. (2)H quadrupolar couplings were measured to determine the motional amplitudes of POPE and POPG acyl chains as a function of temperature. Homogeneously mixed POPE/POPG membranes should give the same quadrupolar couplings for the two lipids, whereas the presence of membrane domains enriched in one of the two lipids should cause distinct (2)H quadrupolar couplings that reflect different chain disorder. At physiological temperature (308 K), we observed no or only small coupling differences between POPE and POPG in the presence of any of the cationic peptides. However, around ambient temperature (293 K), at which gel- and liquid-crystalline phases coexist in the peptide-free POPE/POPG membrane, the peptides caused distinct quadrupolar couplings for the two lipids, indicating domain formation. The broad-spectrum antimicrobial peptide PG-1 ordered ∼40% of the POPE lipids while disordering POPG. The Gram-negative selective PG-1 mutant, IB549, caused even larger differences in the POPE and POPG disorder: ∼80% of POPE partitioned into the ordered phase, whereas all of the POPG remained in the disordered phase. In comparison, TAT rigidified POPE and POPG similarly in the binary membrane at ambient temperature, indicating that TAT does not cause dynamic heterogeneity but interacts with the membrane with a different mechanism. Penetratin maintained the POPE order but disordered POPG, suggesting moderate domain separation. These results provide insight into the extent of domain formation in bacterial membranes and the possible peptide structural requirements for this phenomenon.


Natural developmental variations in leaf and plant senescence in Arabidopsis thaliana.
Leaf senescence is a developmentally regulated process that contributes to nutrient redistribution during reproductive growth and finally leads to tissue death. Manipulating leaf senescence through breeding or genetic engineering may help to improve important agronomic traits, such as crop yield and the storage life of harvested organs. Here, we studied natural variations in the regulation of plant senescence among 16 Arabidopsis thaliana accessions. Chlorophyll content and the proportion of yellow leaves were used as indicator parameters to determine leaf and plant senescence respectively. Our study indicated significant genotype effects on the onset and development of senescence. We selected three late- and five early-senescence accessions for further physiological studies. The relationship between leaf and plant senescence was accession-dependent. There was a significant correlation between plant senescence and the total number of leaves, siliques and plant bolting age. We monitored expression of two senescence marker genes, SAG12 and WRKY53, to evaluate progression of senescence. Our data revealed that chlorophyll content does not fully reflect leaf age, because even fully green leaves had already commenced senescence at the molecular level. Integrating senescence parameters, such as the proportion of senescent leaves, at the whole plant level provided a better indication of the molecular status of the plant than single leaf senescence parameters.


Real ear measures of auditory brain stem response click spectra in infants and adults.
This study investigated spectral differences for auditory brain stem response (ABR) clicks measured in infant and adult ear canals. Click stimuli were presented via insert earphones into the ear canals of 15 adults and 11 infants at 30 and 60 dB nHL. An ER-7C probe microphone fed the stimulus to a spectrum analyzer for analysis of spectral content. Infants had spectral peaks at higher frequencies (mean = 2339.77 Hz) than adults (mean = 1618.75 Hz). The infants also exhibited greater variability in their spectral peaks (SD = 669.37) than did adults (SD = 319.0). The total energy of the clicks was the same in infants' and adults' ears. These results indicate that click stimuli such as those used for ABR testing resonate in infant and adult ears differently, and that the resonance in infants is much more variable than the resonance of adults. These findings provide additional evidence for the need to use infant norms when screening infant hearing using the ABR. Similarly, the findings suggest that infant ABRs may be more correlated with higher frequency behavioral thresholds than those assumed for adults.


Carbonyl Compounds Produced by Vaporizing Cannabis Oil Thinning Agents.
Cannabis use has increased in the United States, particularly the use of vaporized cannabis oil, which is often mixed with thinning agents for use in vaporizing devices. E-cigarette research shows that heated thinning agents produce potentially harmful carbonyls; however, similar studies have not been conducted (1) with agents that are commonly used in the cannabis industry and (2) at temperatures that are appropriate for cannabis oil vaporization. The goal of this study was to determine whether thinning agents used in the cannabis industry produce potentially harmful carbonyls when heated to a temperature that is appropriate for cannabis oil vaporization. Four thinning agents (propylene glycol [PG], vegetable glycerin [VG], polyethylene glycol 400 [PEG 400], and medium chain triglycerides [MCT]) were heated to 230°C and the resulting vapors were tested for acetaldehyde, acrolein, and formaldehyde. Each agent was tested three times. Testing was conducted in a smoking laboratory. Carbonyl levels were measured in micrograms per puff block. Analyses showed that PEG 400 produced significantly higher levels of acetaldehyde and formaldehyde than PG, MCT, and VG. Formaldehyde production was also significantly greater in PG compared with MCT and VG. Acrolein production did not differ significantly across the agents. PG and PEG 400 produced high levels of acetaldehyde and formaldehyde when heated to 230°C. Formaldehyde production from PEG 400 isolate was particularly high, with one inhalation accounting for 1.12% of the daily exposure limit, nearly the same exposure as smoking one cigarette. Because PG and PEG 400 are often mixed with cannabis oil, individuals who vaporize cannabis oil products may risk exposure to harmful formaldehyde levels. Although more research is needed, consumers and policy makers should consider these potential health effects before use and when drafting cannabis-related legislation.


Syrian refugee mothers' knowledge levels of childhood wellness: A program evaluation.
Children are most affected by migration and wars. The health of child asylum seekers is adversely affected due to poor nutrition, malnutrition, insufficient vaccinations, and a lack of preventive health-care services (PHCS). The aim of this study was to determine the knowledge levels of the refugee mothers of child wellness monitoring, child vaccinations, and the importance of breastfeeding before and after implementation of an educational program. A questionnaire consisting of 12 statements about the importance of child wellness monitoring, child vaccinations, and breastfeeding was prepared and translated into Arabic. Refugee mothers were asked to complete the questionnaire before and after the education program. Thirty-one (72.1%) of the 43 participating mothers had their children vaccinated regularly. Vitamin D supplementation was given to 58.1%, and 23.1% were started on iron supplementation. The rate of vitamin D supplementation was higher in the literate mothers (P = 0.010). The least correctly answered statement before the education program was related to iron supplementation (n = 24, 55.8%). The mean (standard deviation) number of correct answers given by the participating mothers to the 12 statements before the seminar was 9.16 (±2.05), this increased to 11.16 (±0.99) after the seminar. The resultes of this study show that refugee mothers' knowledge levels about vitamin D and iron supplementation are quite low. Their knowledge levels can be increased in the short term by providing information in the mothers' native languages.


The extent of gametophytic-sporophytic gene expression in maize.
To determine the extent of gametophytic gene expression and the type of transcription, haploid or haplo-diploid, of the genes, isozymes were used as genetic markers. Fifteen enzymatic systems, including thirty-four isozymes, were studied. The determination of the type of expression of genes coding for multimeric enzymes was based on the comparison of electrophoretic patterns of pollen and of sporophytic tissues from plants heterozygous for electrophoretic mobility: if gene expression in pollen is of a gametophytic (haploid) origin, pollen, unlike the sporophyte, would reveal only the parental homomultimeric bands. The enzymes analyzed can be grouped in three categories according to type of gene expression: i) enzymes present in both pollen and sporophyte, coded by the same gene with haplo-diploid expression; ii) enzymes controlling analogous functions in pollen and sporophyte, coded by different genes, expressed in only one of the two phases; iii) enzymes present in two or more forms in the sporophyte and only in one form in the gametophyte. The data allow the proportion of haplo-diploid gene expression in the loci examined to be estimated at 0.72; 0.22 and 0.06 being the proportions attributable to the sporophytic and gametophytic domains, respectively.


The Piks allele of the NLR immune receptor Pik breaks the recognition of AvrPik effectors of rice blast fungus.
Arms race co-evolution of plant-pathogen interactions evolved sophisticated recognition mechanisms between host immune receptors and pathogen effectors. Different allelic haplotypes of an immune receptor in the host mount distinct recognition against sequence or non-sequence related effectors in pathogens. We report the molecular characterization of the Piks allele of the rice immune receptor Pik against rice blast pathogen, which requires two head-to-head arrayed nucleotide-binding sites and leucine-rich repeat proteins. Like other Pik alleles, both Piks-1 and Piks-2 are necessary and sufficient for mediating resistance. However, unlike other Pik alleles, Piks does not recognize any known AvrPik variants of Magnaporthe oryzae. Sequence analysis of the genome of an avirulent isolate V86010 further revealed that its cognate avirulence (Avr) gene most likely has no significant sequence similarity to known AvrPik variants. Piks-1 and Pikm-1 have only two amino acid differences within the integrated heavy metal-associated (HMA) domain. Pikm-HMA interacts with AvrPik-A, -D, and -E in vitro and in vivo, whereas Piks-HMA does not bind any AvrPik variants. Characterization of two amino acid residues differing Piks-1 from Pikm-1 reveal that Piks-E229Q derived from the exchange of Glu229 to Gln229 in Piks-1 gains recognition specificity against AvrPik-D but not AvrPik-A or -E, indicating that Piks-E229Q partially restores the Pikm spectrum. By contrast, Piks-A261V derived from the exchange of Ala261 to Val261 in Piks-1 retains Piks recognition specificity. We conclude that Glu229 in Piks-1 is critical for Piks breaking the canonical Pik/AvrPik recognition pattern. Intriguingly, binding activity and ectopic cell death induction is maintained between Piks-A261V and AvrPik-D, implying that positive outcomes from ectopic assays might be insufficient to deduce its immune activity against the relevant effectors in rice and rice blast interaction.


Objective assessment of technical surgical skills.
Surgeons are increasingly being scrutinized for their performance and there is growing interest in objective assessment of technical skills. The purpose of this study was to review all evidence for these methods, in order to provide a guideline for use in clinical practice. A systematic search was performed using PubMed and Web of Science for studies addressing the validity and reliability of methods for objective skills assessment within surgery and gynaecology only. The studies were assessed according to the Oxford Centre for Evidence-based Medicine levels of evidence. In total 104 studies were included, of which 20 (19.2 per cent) had a level of evidence 1b or 2b. In 28 studies (26.9 per cent), the assessment method was used in the operating room. Virtual reality simulators and Objective Structured Assessment of Technical Skills (OSATS) have been studied most. Although OSATS is seen as the standard for skills assessment, only seven studies, with a low level of evidence, addressed its use in the operating room. Based on currently available evidence, most methods of skills assessment are valid for feedback or measuring progress of training, but few can be used for examination or credentialing. The purpose of the assessment determines the choice of method.


Synthesis and antimalarial activity of novel side chain modified antimalarial agents derived from 4-aminoquinoline.
Malaria is one of the foremost public health problems in developing countries affecting nearly 40% of the global population. Apart from this, the past two decade's emergence of drug resistance has severely limited the choice of available antimalarial drugs. Furthermore, the general trend emerging from the SAR-studies is that chloroquine resistance does not involve any change to the target of this class of drugs but involves compound specific efflux mechanism. Based on this premise a number of groups have developed short chain analogues of 4-aminoquinoline, which are active against CQ-resistant strains of P. falciparum in in vitro studies. However, these derivatives undergo biotransformation (de-alklyation) significantly affecting lipid solubility of the drug. In view of this background information, we thought that it would be interesting to study the effect of additional lipophilicity and cationic charge at the lateral side chain of 4-aminoquinoline. This prompted us to explore the cationic amino acid conjugates namely, lysine and ornithine of 4-aminoquinoline with a view to achieve improved antimalarial activity and to the best of our knowledge such amino acid conjugates have not been hitherto reported in the literature in the case of 4-aminoquinolines. In the present study, a new series of side-chain modified 4-aminoquinolines have been synthesized and found active against both susceptible and multidrug resistant strains of P. falciparum in vitro and P. yoelli in vivo. The seminal finding of the present study is that a new series of compounds having significantly more activity against CQ resistant parasites has been identified.


Content analysis of work limitation, stanford presenteeism, and work instability questionnaires using international classification of functioning, disability, and health and item perspective framework.
Background. Presenteeism refers to reduced performance or productivity while at work due to health reasons. WLQ-26, SPS-6, and RA-WIS are the commonly used self-report presenteeism questionnaires. These questionnaires have acceptable psychometric properties but have not been subject to structured content analysis that would define their conceptual basis. Objective. To describe the conceptual basis of the three questionnaires using ICF and IPF and then compare the distribution and content of codes to those on the vocational rehabilitation core set. Methods. Two researchers independently linked the items of the WLQ-26, SPS-6, and RA-WIS to the ICF and IPF following the established linking rules. The percentage agreement on coding was calculated between the researchers. Results. WLQ-26 was linked to 62 ICF codes, SPS-6 was linked to 17 ICF codes, and RA-WIS was linked to 74 ICF codes. Most of these codes belonged to the activity and participation domains. All the concepts were classified by the IPF, and the most were rational appraisals within the social domain. Only 12% of codes of the core set for vocational rehabilitation were used in this study to code these questionnaires. Conclusion. The specific nature of work disability that was included in these three questionnaires was difficult to explain using ICF since many aspects of content were not confined. The core set for vocational rehabilitation covered very limited content of the WLQ-26, SPS-6, and RA-WIS.


The insulin-3 gene: lack of a genetic basis for human cryptorchidism.
The etiology of cryptorchidism appears to be multifactorial and related to hormonal and mechanical factors. Recently, the insulin-3 gene (INSL3) was noted to have a role in mouse gubernacular development and testicular descent. Knockout male mice for the INSL3 gene show isolated bilateral cryptorchidism. This phenotype suggests that INSL3 may have a role in the development of human cryptorchidism. Using single strand conformational polymorphism analysis we detected mutations of the INSL3 gene in boys with cryptorchidism. Genomic DNA from 118 boys with cryptorchidism and 48 normal controls were obtained from 3 institutions. Using polymerase chain reaction with INSL3 sequence specific primers DNA fragments were analyzed using single strand conformational polymorphism reactions. Samples with band shifts were re-amplified and sequenced to detect mutations. A single base substitution (G greater than A) causing an amino acid change (missense mutation) was identified in 27 of 118 cryptorchid (23%) samples and 12 of 48 normal (25%) samples. Two other base substitutions did not produce alterations in the amino acid sequence (silent mutations). Although a common polymorphism was detected in the INSL3 gene, no specific mutations were detected in a large population of individuals with cryptorchidism. Therefore, mutations in the coding region of the INSL3 gene are not a common cause of human cryptorchidism.


Cost effectiveness of nurse delivered endoscopy: findings from randomised multi-institution nurse endoscopy trial (MINuET).
To compare the cost effectiveness of nurses and doctors in performing upper gastrointestinal endoscopy and flexible sigmoidoscopy. As part of a pragmatic randomised trial, the economic analysis calculated incremental cost effectiveness ratios, and generated cost effectiveness acceptability curves to address uncertainty. 23 hospitals in the United Kingdom. 67 doctors and 30 nurses, with a total of 1888 patients, from July 2002 to June 2003. Diagnostic upper gastrointestinal endoscopy and flexible sigmoidoscopy carried out by doctors or nurses. Estimated health gains in QALYs measured with EQ-5D. Probability of cost effectiveness over a range of decision makers' willingness to pay for an additional quality adjusted life year (QALY). Although differences did not reach traditional levels of significance, patients in the doctor group gained 0.015 QALYs more than those in the nurse group, at an increased cost of about pound56 (euro59, $78) per patient. This yields an incremental cost effectiveness ratio of pound3660 (euro3876, $5097) per QALY. Though there is uncertainty around these results, doctors are probably more cost effective than nurses for plausible values of a QALY. Though upper gastrointestinal endoscopies and flexible sigmoidoscopies carried out by doctors cost slightly more than those by nurses and improved health outcomes only slightly, our analysis favours endoscopies by doctors. For plausible values of decision makers' willingness to pay for an extra QALY, endoscopy delivered by nurses is unlikely to be cost effective compared with endoscopy delivered by doctors. International standard RCT 82765705.


Paediatric developmental screening: a survey of general practitioners.
A questionnaire survey of 293 general practitioner trainers in England to investigate paediatric screening yielded a response rate of 86%. Paediatric screening sessions were being held by a practice member in the practices of 54% of respondents. In one-third of these practices the practice member was acting in the capacity of clinical medical officer. Of responding trainers 28% held sessions personally and these doctors did not differ significantly from the remainder in terms of sex, seniority, hospital paediatric experience or membership of the Royal College of General Practitioners. About one-third of the doctors holding sessions had spent six months or more working in hospital paediatric departments. First-hand experience of paediatric screening was gained by 60% of the current trainees.Sixty-one per cent of trainers agreed with the view that developmental screening is an appropriate task for all general practitioners, while 71% saw it as an appropriate task for themselves. Eight-six per cent of trainers agreed that doctors should be paid for this service if trained for it, and 56% that they should be paid regardless of training.Comparative figures were determined from a parallel survey of 333 non-training general practitioners of whom 225 (68%) replied. Paediatric screening sessions were held in the practices of 34% of respondents and personally by 21%.It is concluded that there is a high level of interest in paediatric screening among general practitioners, but that there is a need for further expansion in postgraduate paediatric training.


Therapeutic efficacy of the thymidine kinase/ganciclovir system on large experimental gliomas: a nuclear magnetic resonance imaging study.
Contradictory experimental results and human trials have questioned the clinical relevance of the HSVtk/ganciclovir system. To bypass the problem of transfection efficiency, we used a glioma cell line stably expressing the HSVtk gene, which was also fully characterized from gene to protein. We also designed a more clinically relevant experimental protocol, consisting of late GCV delivery on large tumor formations. In short-term studies, histological examination revealed a significant decrease in tumor volume in GCV-treated animals from day 1 or from day 10 after cell inoculation. We observed that late GCV delivery is as efficient as early delivery, probably because GCV can reach tumor cells more easily when neoangiogenesis occurs. In long-term experiments, the survival of treated rats bearing 15-day tumors was improved by 60% compared with C6 control animals. Surprisingly, a 30% survival rate was observed in C6TK control animals. Nuclear magnetic resonance imaging demonstrated, in all surviving animals, a complete regression of tumors without mass effect. These results clearly demonstrate that the HSVtk/GCV system remains a potent therapeutic strategy, even when tested in large tumors, in contrast with the microscopic tumor formations previously reported.


Therapeutic effects of cefotiam and cefazolin on experimental pneumonia caused by Klebsiella pneumoniae DT-S in mice.
The efficacies of several dosage schedules, productive of plasma levels of cefotiam and cefazolin of short and long duration and starting at three levels of cefotiam and cefazolin of short and long duration and starting at three different times (3, 18, and 30h) after infection, were examined in experimental pneumonia caused by Klebsiella pneumoniae DT-S in mice. With each of the multiday regimens there was a large segment of the day when plasma levels fell below assayable concentrations. In all cases, cefotiam proved about eight times as active as cefazolin, indicating that the potent in vitro antibacterial activity of cefotiam was well reflected in the therapeutic effect in this model infection. As judged by the total dose administered, the regimen of cefotiam producing a low but sustained plasma level gave better therapeutic effects than that exhibiting a high but transient plasma level. The cefotiam levels in the plasma of mice that received the regimen effective when initiated at 18 h after infection were less than the expected levels in humans after intravenous infusion of the usual clinical dose.


Transcranial magnetic stimulation in cognitive rehabilitation.
Repetitive transcranial magnetic stimulation (rTMS) can generate an increase or a decrease of neuronal excitability, which can modulate cognition and behaviour. Transcranial magnetic stimulation-induced cortical changes have been shown to result in neural plasticity. Thus, TMS provides an important opportunity to gain more insight into the mechanisms responsible for the remarkable flexibility of the central nervous system. The aim of this review was to cover the topics that could be useful when using TMS in the cognitive rehabilitation field after brain damage. The basic TMS principles are introduced, together with the clinical application for diagnosis and prognosis, the biological aspects, and the use in cognitive neuroscience studies. Finally, several hypotheses are discussed to explain the likely mechanisms induced by TMS that favour the recovery of a function after brain damage and cause the adult brain to undergo plasticity. The possibility of non-invasively interacting with the functioning of the brain and its plasticity mechanisms - a possibility that may eventually lead to cognitive and behavioural modifications - opens new and exciting scenarios in the cognitive neurorehabilitation field.


Fitting the anophthalmic socket: achieving cosmesis with comfort.
In attempting to achieve acceptable cosmesis while maintaining comfort for the anophthalmic patient, communication between the ocularist and oculoplastic surgeon is of paramount importance. The ocularist must realize his limitations in correcting deformities prosthetically and must communicate these limitations to the ophthalmologist when surgical alternatives are being considered.


Synthesis and reactivity of the unsaturated trinuclear phosphanido complex [(C6F5)2Pt(μ-PPh2)2Pt(μ-PPh2)2Pt(PPh3)].
The reaction of [NBu(4)][(C(6)F(5))(2)Pt(μ-PPh(2))(2)Pt(μ-PPh(2))(2)Pt(O,O-acac)] (48 VEC) with [HPPh(3)][ClO(4)] gives the 46 VEC unsaturated [(C(6)F(5))(2)Pt(1)(μ-PPh(2))(2)Pt(2)(μ-PPh(2))(2)Pt(3)(PPh(3))](Pt(2)-Pt(3)) (1), a trinuclear compound endowed with a Pt-Pt bond. This compound displays amphiphilic behavior and reacts easily with nucleophiles L, yielding the saturated complexes [(C(6)F(5))(2)Pt(II)(μ-PPh(2))(2)Pt(II)(μ-PPh(2))(2)Pt(II)(PPh(3))L] [L = PPh(3) (2), py (3)]. The reaction with the electrophile [Ag(OClO(3))PPh(3)] affords the adduct 1·AgPPh(3), which evolves, even at low temperature, to a mixture in which [(C(6)F(5))(2)Pt(III)(μ-PPh(2))(2)Pt(III)(μ-PPh(2))(2)Pt(II)(PPh(3))(2)](2+)(Pt(III)-Pt(III)) and 2 (plus silver metal) are present. The nucleophilic nature of 1 is also demonstrated through its reaction with cis-[Pt(C(6)F(5))(2)(THF)(2)], which results in the formation of [Pt(4)(μ-PPh(2))(4)(C(6)F(5))(4)(PPh(3))] (4). The structure and NMR features indicate that 1 can be better considered as a Pt(II)-Pt(III)-Pt(I) complex instead of a Pt(II)-Pt(II)-Pt(II) derivative. Theoretical calculations (density functional theory) on similar model compounds are in agreement with the assigned oxidation states of the metal centers. The strong intermetallic interactions resulting in a Pt(2)-Pt(3) metal-metal bond and the respective bonding mechanism were verified by employing a multitude of computational techniques (natural bond order analysis, the Laplacian of the electron density, and localized orbital locator profiles).


Self-specific processing in the meditating brain: a MEG neurophenomenology study.
Self-specific processes (SSPs) specify the self as an embodied subject and agent, implementing a functional self/nonself distinction in perception, cognition, and action. Despite recent interest, it is still undetermined whether SSPs are all-or-nothing or graded phenomena; whether they can be identified in neuroimaging data; and whether they can be altered through attentional training. These issues are approached through a neurophenomenological exploration of the sense-of-boundaries (SB), the fundamental experience of being an 'I' (self) separated from the 'world' (nonself). The SB experience was explored in collaboration with a uniquely qualified meditation practitioner, who volitionally produced, while being scanned by magnetoencephalogram (MEG), three mental states characterized by a graded SB experience. The results were then partly validated in an independent group of 10 long-term meditators. Implicated neural mechanisms include right-lateralized beta oscillations in the temporo-parietal junction, a region known to mediate the experiential unity of self and body; and in the medial parietal cortex, a central node of the self's representational system. The graded nature as well as the trainable flexibility and neural plasticity of SSPs may hold clinical implications for populations with a disturbed SB.


An Anaphylactoid Reaction to Rattlesnake Envenomation in a Pediatric Patient.
Rattlesnake envenomation causes a spectrum of symptoms and signs, many of which closely resemble an anaphylactic reaction. Development of airway compromise after a snake bite to an extremity is unusual, but has been previously reported. The majority of such reports detail a history of probable sensitization from previous contact with snakes or snake venom. Few reports exist without such previous contact; no reports are of pediatric cases. Here we present a 14-year-old boy with no history of prior snake exposure who presents with an anaphylactoid reaction after a rattlesnake bite.


Adsorption and degradation of doxorubicin from aqueous solution in polypropylene containers.
The purpose of this study was to examine doxorubicin adsorption in polypropylene containers as a function of pH and drug concentration based on anecdotal evidence of such adsorption. Doxorubicin loss was first examined in high-performance liquid chromatography (HPLC) glass inserts by UV absorbance to determine appropriate pH and time durations for subsequent analysis. Doxorubicin loss was then investigated in polypropylene microcentrifuge tubes at different pH values and starting drug concentration at 37°C over 48 h using HPLC with fluorescent detection. Doxorubicin concentrations was essentially constant in HPLC glass inserts at pH 4.8 up to 12 h but declined 5% at pH 7.4 by 3 h. The percent doxorubicin adsorption was calculated in polypropylene microcentrifuge tubes from extrapolations to zero time and was the least at pH 4.8, but increased with pH values 6.5 and 7.4, and decreased with drug concentration to reach a maximum adsorption of 45% in 2.0 μg/mL at pH 7.4 and 37°C. Degradation rate constants, ranging from 0.0021 to 0.019 h(-1), also increased with pH in these studies. Determinations of low amounts of doxorubicin in polypropylene containers at pH 7.4 may be underestimated if adsorption and degradation issues are not taken into account.


The mitogen-activated protein kinase kinase 4 has a pro-oncogenic role in skin cancer.
The mitogen-activated protein kinase (MAPK) kinase 4 (MKK4) is a nonredundant component of stress-activated MAPK signaling modules. Its function in tumorigenesis remains highly controversial with some studies indicating that MKK4 is a tumor suppressor, whereas others have reported a pro-oncogenic role. To clarify the role of MKK4 in cancer, we have created a novel mouse model to test the effect of the specific loss of MKK4 in the epidermis on the formation of papillomas caused by activated ras mutation. We have discovered that skin-specific MKK4-deficient mice are resistant to carcinogen-induced tumorigenesis. One mechanism by which MKK4 promotes cell proliferation and the formation of tumors is by increasing epidermal growth factor receptor expression through the c-Jun NH(2)-terminal protein kinase/c-Jun signaling pathway. Together, our results provide the first genetic demonstration that MKK4 is essential to mediate the oncogenic effect of Ras in vivo, thereby validating MKK4 as a potential drug target for cancer therapy.


Association of decreased levels of lipopolysaccharide-binding protein with OKN-007-induced regression of tumor growth in an F98 rat glioma model.
High-grade gliomas, such as glioblastoma (GBM), are devastating tumors with a very poor prognosis. Previously the authors have found that the nitrone compound OKN-007 (OKlahoma Nitrone 007; or disodium 4-[(tert-butyl-imino) methyl] benzene-1,3-disulfonate N-oxide) is effective against high-grade gliomas in various GBM rodent and human xenograft models. The purpose of the present study was to assess the levels of the lipopolysaccharide-binding protein (LBP) in rodent gliomas treated with OKN-007 as well as determine the expression of LBP in human gliomas. Microarray analysis was done to assess altered gene expression following OKN-007 administration in an F98 glioma model. An enzyme-linked immunosorbent assay was incorporated to assess LBP levels in glioma tissues, as well as blood serum, comparing results in OKN-007-treated and untreated tumor-bearing animals. Immunohistochemistry was used to assess LBP levels in varying grades of human glioma tissue sections. Upon further assessment of gene expression fold changes in F98 gliomas in rats that received or did not receive OKN-007, it was found that the gene for LBP was significantly downregulated by OKN-007. Further investigation was done to see whether levels of LBP were affected by OKN-007 treatment in F98 gliomas. It was found that LBP could be detected not only in glioma tissue but also in blood serum of F98 glioma-bearing rats and that OKN-007 decreased the levels of LBP. It was also found that LBP levels are highly expressed in human high-grade glioma tissues. LBP could potentially be used as a serum diagnostic marker of treatment response in high-grade gliomas.


Long-term affinity modification on poly(dimethylsiloxane) substrate and its application for ELISA analysis.
Poly(dimethylsiloxane) (PDMS) possesses many advantages, such as biocompatibility and high oxygen permeability, which makes it an attractive material for fabricating biodevices. Creating an affinity surface with long-term stability and reactivity for biomolecular interactions on a PDMS substrate, however, is difficult due to its inherent hydrophobicity. In this study, an affinity surface on a PDMS substrate with long-term hydrophilicity and affinity reactivity is reported. This modification is composed of two parts. The bottom part is made of polyelectrolyte multilayers and is capable of providing long-term hydrophilic stability. The top part consists of three protein layers, bovine serum albumin (BSA), anti-BSA, and protein G, and offers an affinity surface for antibody binding and, more importantly, provides favorable orientation and minimum nonspecific binding. The chemical modification for the different stages was monitored by atomic force microscopy (AFM), attenuated total reflection Fourier transform infrared spectroscopy (ATR-FT-IR), and contact angle and fluorescence measurements. A long-term PDMS immunodevice (LPID) based on polyelectrolyte multilayers and protein layers was fabricated and applied to the detection of transforming growth factor beta (TGF-beta) protein in mouse serum by the enzyme-linked immunosorbent assay (ELISA) method. Results show that a linear calibration curve was obtained in the concentration range from 500 to 15.125 pg/mL, and the relative standard deviation was less than 3%. Also, the amount of TGF-beta spiked in mouse serum was precisely determined. Results indicate that the modified surface was hydrophilic and reactive to biospecies up to more than 7 days in its dry form. Moreover, the blocking reagent used to reduce nonspecific binding was found to be not necessary for the LPID. Thus, the reported method is expected to hold a great potential for fabricating PDMS-based affinity devices such as protein chips.


Knee laxity after anterior tibial eminence fracture in children: A 35-case series.
Anterior tibial eminence (ATE) fractures are characterized by avulsion of the anterior cruciate ligament insertion. The aim of our study was to evaluate the long-term incidence of laxity and instability in the aftermath of these fractures. The secondary objective was to identify factors for instability. ATE fracture in children is responsible for laxity and instability in the medium and long term. This retrospective, single-center study included 35 isolated fractures of the tibial intercondylar eminence during skeletal growth between January 2006 and January 2020. Analysis comprised demographics, laxity measured by GNRB™, range of motion and IKDC and Lysholm scores. Clinical reassessment was performed in 24 patients, the other 11 being interviewed by telephone. Mean laxity on GNRB™ was 1.46mm, and 3 patients had>3mm differential with respect to the healthy knee. Mean IKDC score was 92.2 and mean Lysholm score 93.1. Four patients showed instability, 2 of whom required surgical management. There was no significant difference in occurrence of laxity according to fracture type or reduction quality. Mean follow-up was 5.9years (range, 1.1-14.8). Our clinical and functional results were in accordance with the literature. The long-term clinical results were satisfactory. ATE fractures require long-term follow-up to screen for instability and laxity on GNRB™. IV, retrospective study.


Growth hormone attenuates the acute-phase response to thermal injury.
To determine the effects of growth hormone (GH) on the hepatic acute-phase response (APR) in a burned rat model. Laboratory. Male Sprague-Dawley rats (weight, 300-350 g). Rats underwent a 40% total body surface area burn injury and received GH or saline solution daily by subcutaneous injection. Unburned rats served as controls. Hepatic messenger RNA (mRNA) expression and serum levels of alpha 1-acid glycoprotein and albumin were determined 2, 7, and 14 days after injury. The serum alpha 1-acid glycoprotein levels in GH-treated animals did not increase on days 2 and 7, whereas saline-treated animals showed a major increase. Hepatic mRNA expression increased dramatically on day 2 for burned groups; however, the mRNA pool levels of GH-treated animals showed a faster rate of decline to control levels on days 7 and 14. The albumin mRNA pool levels of GH-treated and control animals did not show significant differences, whereas the negative APR, indicated by loss of albumin mRNA, was more pronounced on day 7 in the saline-treated animals. By day 14, mRNA levels were comparable in all 3 groups. Growth hormone attenuated the positive APR, as indicated by a decrease in alpha 1-acid glycoprotein expression and production, and prevented the negative APR, as seen by an absence of a decline of albumin mRNA pool levels and serum concentration. We conclude that the beneficial effects of GH on thermal injury may be due in part to a modification of the APR.


Historically and currently used Dechloranes in the sediments of the Great Lakes.
Dechlorane (mirex), Dechlorane Plus (DP), Dechlorane 602 (Dec602), Dechlorane 603 (Dec603), Dechlorane 604 (Dec604), and Chlordecone (Kepone) were analyzed in 16 sediment cores collected in 2007 from the Great Lakes of North America. Results show that Lake Ontario sediments have accumulated mirex, DP, Dec602 and Dec604 in amounts 1 to 2 orders of magnitude higher than other Great Lakes. The chemical inventory decreases log-linearly with increasing latitude (N) and longitude (W) of the sampling locations, but Lake Ontario sites are outliers in the regression against latitude. The regression analyses suggest differences among the analytes with regard to source impact and long-range transport behavior. Temporal trends of input differ among lakes. Most sites in Lake Superior are still receiving increasing fluxes of DP and Dec602, while these have been declining in Lake Ontario from the peak around 1990. The relative abundance of the two DP isomers, represented by f(syn), increases with increasing distance from the potential discharge source in Niagara Falls, NY, suggesting the anti-DP isomer is more vulnerable to degradation during long-range atmospheric transport. Kepone was not detected in the sediments of Lakes Ontario, Erie, and Michigan.


Long-lasting plasma membrane permeabilization in mammalian cells by nanosecond pulsed electric field (nsPEF).
The barrier function of plasma membrane in nsPEF-exposed mammalian cells was examined using whole-cell patch-clamp techniques. A specialized setup for nsPEF exposure of individual cells in culture was developed and characterized for artifact-free compatibility with the patch-clamp method. For the first time, our study provides experimental evidence that even a single 60-ns pulse at 12 kV/cm can cause a profound and long-lasting (minutes) reduction of the cell membrane resistance (R(m)), accompanied by the loss of the membrane potential. R(m) measured in GH3, PC-12, and Jurkat cells (but not in HeLa cells) in 80-120 s after nsPEF exposure was decreased about threefold, and its gradual recovery could take 15 min. Multiple pulses enhanced permeabilization, for example, R(m) in GH3 cells fell about 10-fold after a train of five pulses. Within studied limits, permeabilization did not depend on the presence of Ca(2+), Mg(2+), K(+), Cs(+), Cd(2+), EGTA, tetraethylammonium, or 4-aminopyridine in the pipette or bath solutions. Our results supported theoretical model predictions of plasma membrane poration by nsPEF. However, the extended decrease in R(m), assumed to be related to the life span of the pores, and different nsPEF sensitivity of individual cell lines have yet to be explained. The phenomenon of long-lived membrane permeabilization provides new insights on the nature of nsPEF-opened conductance pores and on molecular mechanisms that underlie nsPEF bioeffects.


Clinical correlates of oral impacts on daily performances.
The aim of this study was to investigate the associations between oral health measures and oral health-related quality of life as captured by OIDP (oral impacts on daily performances). The study was performed in three dental clinics in Sweden and included 204 patients, 43.8% men and 56.2% women (aged 20-86 years), consecutively recruited in connection with their routine dental examination. The patients were interviewed using the OIDP followed by a clinical examination. Four bite-wing radiographs were taken in two of the clinics (n = 154). A self-administered questionnaire provided information about socio-economic data. Subjects >or=60 years had significantly more missing teeth, lesser maximal jaw opening, higher number of sites with alveolar bone loss and proportionally more filled teeth than younger individuals. Impacts related to the oral health that affected their daily life were reported in 39.7%. Multivariate logistic regressions analysis showed that missing teeth (>or=10) and a limited jaw opening (<40 mm) were significantly associated with having one or more impact as measured with the OIDP [odds ratio (OR) 6.50, 95% CI 1.48-28.43 and OR 2.87, 95% CI 1.03-7.96, respectively]. Individuals with diminished functional oral health status (missing teeth and limited jaw opening) had significantly more often one or more oral impacts on daily life than those with fewer than 10 missing teeth and a jaw opening >or=40 mm. The OIDP instrument may be valuable for use in routine dental check-ups in patients with related problems to determine possible oral impacts on daily life.


The development of the rank-order stability of the Big Five across the life span.
Several studies have suggested that the rank-order stability of personality increases until midlife and declines later in old age. However, this inverted U-shaped pattern has not consistently emerged in previous research; in particular, a recent investigation implementing several methodological advances failed to support it. To resolve the matter, we analyzed data from two representative panel studies and investigated how certain methodological decisions affect conclusions regarding the age trajectories of stability. The data came from Australia (N = 15,465; Study 1) and Germany (N = 21,777; Study 2), and each study included four waves of personality assessment. We investigated the life span development of the rank-order stability of the Big Five for 4-, 8-, and 12-year intervals. Whereas Study 1 provided strong evidence for an inverted U-shape with rank-order stability declining past age 50, Study 2 provided more mixed results that nonetheless generally supported the inverted U-shape. This developmental trend held for single personality traits as well as for the overall pattern across traits; and it held for all three retest intervals-both descriptively and in formal tests. Additionally, we found evidence that health-related changes accounted for the decline in rank-order stability in older age. This suggests that if analyses are implicitly conditioned on health (e.g., by excluding participants with missing data on later waves), the decline in stability in old age will be underestimated or even missed. Our results provide further evidence for the inverted U-shaped age pattern in personality stability development but also extend knowledge about the underlying processes. (PsycInfo Database Record (c) 2022 APA, all rights reserved).


Factors Involved in the Food Choices of Diners in a Kibbutz Communal Dining Room Buffet: A Qualitative Study.
Improving nutrition improves health outcomes. Eating in a catering system may provide an environment for promoting healthy dietary choices. To map the factors that shape the food choices of diners who routinely eat in catering systems, we collected and analyzed qualitative data about diners' perceptions of their food choices in communal dining rooms in three kibbutzim in Israel. From May to July 2014, we conducted in-depth, semi-structured, face-to-face interviews with 13 diners who ate at least three lunches per week in the kibbutz's dining room. Data analysis followed thematic analysis principles. Two categories of themes emerged from the interviews. In the personal context category, the themes identified were eating as a task and attempts to control one's eating. In the contextual aspects of eating in the catering system category, themes identified were eating in the dining room as a default, the characteristics of the food served, routine, and personal versus public aspects. The sub-theme of the diners' freedom of choice emerged in the two categories of themes. Diners' wishes of maintaining their freedom of choice may be an important contribution to the debate of whether catering systems should provide only healthy foods, which may jeopardize diners' freedom of choice.


Targeting of cytotoxic luteinizing hormone-releasing hormone analogs to breast, ovarian, endometrial, and prostate cancers.
Targeted chemotherapy is a modern approach aimed at increasing the efficacy of systemic chemotherapy and reducing its side effects. The peptide receptors expressed primarily on cancerous cells can serve as targets for a selective destruction of malignant tumors. Binding sites for LHRH (now known in genome and microarray databases as GNRH1), were found on 52% of human breast cancers, about 80% of human ovarian and endometrial cancers, and 86% of human prostatic carcinoma specimens. Because LHRH receptors are not expressed on most normal tissues, they represent a specific target for cancer chemotherapy with antineoplastic agents linked to an LHRH vector molecule. To test the efficacy of targeted chemotherapy based on LHRH analogs, we recently developed a cytotoxic analog of LHRH, designated AN-152, which consists of [D-Lys6]LHRH covalently linked to one of the most widely used chemotherapeutic agents, doxorubicin (DOX). In addition, we designed and synthesized a highly active derivative of DOX, 2-pyrrolino-DOX (AN-201), which is 500-1000 times more potent than DOX in vitro. AN-201 is active against tumors resistant to DOX, and noncardiotoxic. As in the case of DOX, AN-201 was coupled to carrier peptide [D-Lys6]LHRH to form a superactive targeted cytotoxic LHRH analog, AN-207. Both AN-152 and AN-207 can effectively inhibit the growth of LHRH receptor-positive human breast, ovarian, endometrial, and prostate cancers xenografted into nude mice. DOX-containing cytotoxic LHRH analog AN-152 is scheduled for clinical phase I/IIa trials in patients with advanced ovarian and breast cancers in 2005.


Array of stacked leaky-wave antennas in groove gap waveguide technology.
The design of an array of stacked leaky-wave antennas in groove gap waveguide technology is presented in this work. The proposed array is formed by simply stacking a number of leaky-wave antennas one on top of the other and feeding all of them with uniform amplitude and phase. The inter-element distance is studied in order to avoid grating lobes and to maximize the directivity. A feeding network based on vertical coupling is designed, where the input port feeds the bottom element, and then the energy is equally coupled to the other elements. To obtain maximum directivity the phase is corrected at each element separately. The central frequency of the proposed design is 28 GHz. With this technique of stacking the elements a pencil beam is achieved, i.e. the radiated energy is focalized in the two main planes. The designed array with four elements achieves an enhancement of + 5 dB, reaching 24.5 dBi of directivity in comparison to 19.6 dBi of directivity of the single leaky-wave antenna made in this technology. A prototype is manufactured and measured and its results are presented and compared with the simulations.


The science of space weather.
The basic physics underpinning space weather is reviewed, beginning with a brief overview of the main causes of variability in the near-Earth space environment. Although many plasma phenomena contribute to space weather, one of the most important is magnetic reconnection, and recent cutting edge research in this field is reviewed. We then place this research in context by discussing a number of specific types of space weather in more detail. As society inexorably increases its dependence on space, the necessity of predicting and mitigating space weather will become ever more acute. This requires a deep understanding of the complexities inherent in the plasmas that fill space and has prompted the development of a new generation of scientific space missions at the international level.


Pseudomonas aeruginosa: evolution of antimicrobial resistance and implications for therapy.
Pseudomonas aeruginosa is a formidable pathogen in the infection arena. It is able to easily adapt to the environment which it inhabits and can also colonize and invade the human host to cause serious infections. In 2011, it was responsible for 7.1% of all health care-associated infection in the United States. The morbidity and mortality of both blood stream infections and ventilator-associated pneumonia are significant. On a global scale, we have seen the development of not only multidrug resistance but also extensive and pan drug resistance in this organism. This is often associated with limited clonal types of which we now have epidemiological evidence of spread. With this has come reduced antibiotic treatment options. Consideration of antibiotic infusions, combination therapy, and inhalational therapy has occurred in an attempt to gain the upper ground. Gram-negative resistance has appropriately been described as a global emergency.


Consequences of Epithelial Inflammasome Activation by Bacterial Pathogens.
Inflammasome signaling impinges on the activation of inflammatory caspases (i.e., caspase-1 and caspase-4/5/11) and endows host cells with a sentinel system to sense microbial intrusion and thereby initiate appropriate immune responses. Lately, it has become evident that mammalian inflammasome-dependent responses to infection are not confined solely to cells of hematopoietic origin. Epithelial cells that line the body's mucosal surfaces use inflammasome signaling to sense and counteract pathogenic microorganisms that compromise barrier integrity. Many of the molecular mechanisms of epithelial inflammasome signaling remain unexplored. However, it now seems clear that epithelial inflammasome activation has a profound impact both on the infected cell itself and on its ability to communicate with other cell types of the mucosa. Here, we summarize current knowledge regarding the output of epithelial inflammasome activation during bacterial infection. Well-established downstream effects include epithelial cell death, release of soluble mediators, and subsequent recruitment of effector cell types, including NK cells, mast cells, and neutrophils, to sites of mucosal infection. We discuss the implications of recent findings for antibacterial defense in the mucosa and sketch out areas for future exploration.


MORC2 regulates DNA damage response through a PARP1-dependent pathway.
Microrchidia family CW-type zinc finger 2 (MORC2) is a newly identified chromatin remodeling enzyme with an emerging role in DNA damage response (DDR), but the underlying mechanism remains largely unknown. Here, we show that poly(ADP-ribose) polymerase 1 (PARP1), a key chromatin-associated enzyme responsible for the synthesis of poly(ADP-ribose) (PAR) polymers in mammalian cells, interacts with and PARylates MORC2 at two residues within its conserved CW-type zinc finger domain. Following DNA damage, PARP1 recruits MORC2 to DNA damage sites and catalyzes MORC2 PARylation, which stimulates its ATPase and chromatin remodeling activities. Mutation of PARylation residues in MORC2 results in reduced cell survival after DNA damage. MORC2, in turn, stabilizes PARP1 through enhancing acetyltransferase NAT10-mediated acetylation of PARP1 at lysine 949, which blocks its ubiquitination at the same residue and subsequent degradation by E3 ubiquitin ligase CHFR. Consequently, depletion of MORC2 or expression of an acetylation-defective PARP1 mutant impairs DNA damage-induced PAR production and PAR-dependent recruitment of DNA repair proteins to DNA lesions, leading to enhanced sensitivity to genotoxic stress. Collectively, these findings uncover a previously unrecognized mechanistic link between MORC2 and PARP1 in the regulation of cellular response to DNA damage.


A whole-organ regenerative medicine approach for liver replacement.
The therapy of choice for end-stage liver disease is whole-organ liver transplantation, but this option is limited by a shortage of donor organs. Cell-based therapies and hepatic tissue engineering have been considered as alternatives to liver transplantation, but neither has proven effective to date. A regenerative medicine approach for liver replacement has recently been described that includes the use of a three-dimensional organ scaffold prepared by decellularization of xenogeneic liver. The present study investigates a new, minimally disruptive method for whole-organ liver decellularization and three different cell reseeding strategies to engineer functional liver tissue. A combination of enzymatic, detergent, and mechanical methods are used to remove all cells from isolated rat livers. Whole-organ perfusion is used in a customized organ chamber and the decellularized livers are examined by morphologic, biochemical, and immunolabeling techniques for preservation of the native matrix architecture and composition. Three different methods for hepatocyte seeding of the resultant three-dimensional liver scaffolds are evaluated to maximize cell survival and function: (1) direct parenchymal injection, (2) multistep infusion, or (3) continuous perfusion. The decellularization process preserves the three-dimensional macrostructure, the ultrastructure, the composition of the extracellular matrix components, the native microvascular network of the liver, and the bile drainage system, and up to 50% of growth factor content. The three-dimensional liver matrix reseeded with the multistep infusion of hepatocytes generated ∼90% of cell engraftment and supported liver-specific functional capacities of the engrafted cells, including albumin production, urea metabolism, and cytochrome P450 induction. Whole-organ liver decellularization is possible with maintenance of structure and composition suitable to support functional hepatocytes.


NO modulates fetoplacental blood flow distribution and whole body oxygen extraction in fetal sheep.
It is unknown if nitric oxide (NO) influences the relative level of the left (LV) and right ventricular (RV) outputs, the blood flow distribution between the body and placenta, or whole body O2 extraction and O2 consumption in the fetus. To address these questions eight fetal lambs were chronically instrumented at 128-134 days gestation (term 147 days), and blood flows were measured with radioactive microspheres 3-4 days later at baseline and after inhibition of NO synthesis with N omega-nitro-L-arginine (L-NNA, 10 and 25 mg/kg i.v.). L-NNA progressively reduced the combined ventricular output (P < 0.005) but did not alter the relative levels of the LV and RV outputs. Fetal body blood flow fell by 31% after 10 mg/kg L-NNA (P < 0.005), but a reduction in placental blood flow (P < 0.005) was smaller (20%) and not observed until 25 mg/kg L-NNA. Whole body O2 extraction increased by 71% after 10 mg/kg L-NNA (P < 0.005) and did change further at 25 mg/kg L-NNA, whereas whole body O2 consumption rose by 15% at 10 mg/kg L-NNA (P < 0.05) and returned to baseline at 25 mg/kg L-NNA. These results suggest that, as well as reducing the combined ventricular output, inhibition of fetal NO synthesis redistributes systemic blood flow toward the placenta and increases fetal body O2 extraction. The latter initially increases whole body O2 consumption and then maintains it at near baseline levels after a fall in placental perfusion.


Expectations of Middle aged Women for a life of Happiness to the Spouse.
This study was to define and analyze the expectations of middle-aged woman to their spouse. A sample of 39 middle-aged women was recruited from the workplace, religious organization and acquaintances. The survey was conducted with participants using self-reported questionnaire. The data were analyzed using Q methodology. There were four different types of expectations of middle-aged women. First type was "Collaborative and self-development type," meaning that women developed self and were collaborated with their family or spouses. The second type was "Maintain their own life" meaning that women recognized and maintained their expertise without interference from their spouses. The third one was "Respect for the inner value" meaning that women gave continued respect and understanding their inner value. The last one was "Realistic acceptance" meaning that women kept the current situation without implementing new self-development type. There is a subjective structure within the types of expectations of middle-aged women for a life of happiness to the spouse.


[Biological activity of a new Soviet natural inducer, double-stranded RNA].
Antiviral activity of a two-spiral RNA (ts RNA), a new natural interferon inductor was studied. It was shown that ts RNA extracted from a phage infected E. coli culture was an active inductor of interferon and resistance to infection with the forestspring encephalitis virus experimental animals. In experiments on 10-12 g mice ts RNA administered in a dose of 50 micrograms/mouse 6 hours after the infection induced up to 1280 units/ml of the serum interferon. When the inductor was administered repeatedly, the experimental animals developed hyporeactivity resulting in a marked decrease in interferon production after the 3rd subsequent injection. The most pronounced effect with respect to the forest-spring encephalitis virus was observed when the inductor was administered intraperitoneally in a dose of 20 micrograms/mouse 4 hours before the infection. The protective effect was less pronounced when the inductor was administered 24 and 48 hours before the infection. A two-fold administration of the inductor did not increase the antiviral effect. When the inductor was administered in a dose of 100 micrograms 14 days before the infection, the animals showed an increase in the nonspecific resistance to the infection resulting in a marked antiviral effect.


Recurrent laryngeal nerve diameter increases during thyroidectomy.
Temporary recurrent laryngeal nerve (RLN) palsy after thyroidectomy is usually due to a neurapraxia. Voice changes after thyroidectomy, in the absence of RLN palsy, are common. We postulated that this is due to edema and consequent increased diameter of the RLN during thyroidectomy. The aim of this study was to document changes in the diameter of the RLN during surgery. A consecutive series of 110 RLNs was prospectively analyzed in 75 patients. The RLN was measured on identification and removal of the lobe. Electromyogram (EMG) amplitudes were recorded concurrently after neurostimulation. Univariate and multivariate analyses were performed. The mean increase in RLN diameter was 0.71 from 1.95 mm (P < 000.1). Right and left RLNs were similar diameter. On univariate and multivariate analysis, factors predictive of increased RLN diameter were increasing age (P = 0.04) and increased difference in EMG amplitude (P = 0.01). There was a mean increased EMG amplitude of 101 from 493.5 μV (P < 0.001). On univariate and multivariate analyses, factors predictive of increased difference in EMG amplitude were decreasing age (P = 0.01) and increased difference in RLN diameter (P = 0.02). There was a statistically significant positive relationship between increased diameter of RLN and increased EMG amplitude (R(2) = 0.04). The temporary RLN palsy rate was 3%. The small numbers of temporary RLN palsies prevent correlation with increased diameter of RLN. However, this may explain voice changes after thyroidectomy without RLN palsy. The observed increase in RLN diameter is probably due to edema; its cause is unknown. The increased EMG amplitude seen with increasing RLN diameter may reflect increased excitability of ionic channels in neurons.


Urinary biotin analogs increase in humans during chronic supplementation: the analogs are biotin metabolites.
In human subjects, the metabolic origins of bisnorbiotin and biotin sulfoxide were determined by measuring the urinary excretion of each after chronic administration of large oral doses of biotin. For 2 wk, 14 adult volunteers consumed 1,200 micrograms of biotin per day, an amount approximately 20 times the daily dietary intake. With the use of a high-performance liquid chromatography/avidin-binding assay in untimed urine samples obtained before the first dose of biotin and after the 14th dose, concentrations of biotin, bisnorbiotin, and biotin sulfoxide were measured. Excretion was expressed as concentration ratios to urinary creatinine. Bisnorbiotin and biotin sulfoxide excretion increased 85-fold (P < 0.0001) and 114-fold (P < 0.0001), respectively. The molar percentages of bisnorbiotin and biotin sulfoxide decreased from 28 to 14% (P = 0.006) and from 9 to 5% (P = 0.017), respectively. These data provide evidence that the bisnorbiotin and biotin sulfoxide found in human urine are biotin metabolites. Furthermore, we infer that chronic consumption of large amounts of biotin does not substantially saturate the human biotin pathways of biotransformation.


In situ nitrogen doping of TiO2 by plasma enhanced atomic layer deposition for enhanced sodium storage performance.
Nitrogen-doped TiO2 is in situ synthesized by plasma enhanced atomic layer deposition on carbon nanotubes (N-TiO2/CNTs). The prepared N-TiO2/CNT nanocomposites are employed as anode materials for sodium ion batteries. The specific capacity of an N-TiO2/CNT electrode is 1.45 times higher than that of a TiO2/CNT electrode at a current density of 50 mA g-1 after 200 cycles. It is demonstrated that N-TiO2/CNTs are more effective in an electrochemical Na cell due to the enhanced kinetics which results from the nitrogen-doping and the amorphous feature.


More than a virus: a qualitative study of the social implications of hepatitis B infection in China.
China has the largest absolute number of people living with hepatitis B with up to 300,000 people estimated to die each year from hepatitis B related diseases. Despite advances in immunisation, clinical management, and health policy, there is still a lack of accessible and affordable health care for people with hepatitis B. Through in-depth interviews, this study identifies the personal, social and economic impact of living with hepatitis B and considers the role of stigma and discrimination as barriers to effective clinical management of the disease. Semi-structured qualitative interviews were held with 41 people living with hepatitis B in five Chinese cities. Participants were recruited through clinical and non-government organisations providing services to people with hepatitis B, with most (n = 32) being under the age of 35 years. People living with hepatitis B experience the disease as a transformative intergenerational chronic infection with multiple personal and social impacts. These include education and employment choices, economic opportunities, and the development of intimate relationships. While regulations reducing access to employment and education for people with hepatitis B have been repealed, stigma and discrimination continue to marginalise people with hepatitis B. Effective public policy to reduce morbidity and mortality associated with hepatitis B needs to address the lived impact of hepatitis B on families, employment and educational choices, finances, and social marginalisation.


Augmenting serotonin neurotransmission with citalopram modulates emotional expression decoding but not structural encoding of moderate intensity sad facial emotional stimuli: an event-related potential (ERP) investigation.
Antidepressants targeting the serotonergic system have been shown to modulate biases in emotional processing. The effects of serotonergic modulation on the temporal course of emotional processing (accruing within milliseconds) are unknown. Furthermore, it is unknown how serotonin affects different stages of facial emotional processing. The current study investigated the effects of acute serotonin augmentation on event-related potential (ERP) measures associated with 'structural encoding' (N170) and emotion 'expression decoding' (N250 and a late slow-wave positive potential [LPP]) of happy and sad facial stimuli, relative to neutral facial stimuli. The study was a double-blind, placebo-controlled, cross-over design, in which 14 healthy male participants completed a facial recognition task under two acute treatment conditions: 1) placebo (PLB) and 2) 20 mg citalopram (CIT). ERP recording were conducted while subjects viewed neutral, happy and sad facial stimuli. Findings indicated that under PLB, the N170 was not modulated by valence (happy or sad versus neutral), but the N250 and LPP were enhanced for processing happy (relative to neutral) faces. Citalopram had no effect on the N170, but it enhanced the LPP for processing sad (relative to neutral) faces. These findings suggest that serotonin enhancement has selective and temporal effects on emotional face processing, with evidence for modulating processes associated with 'expression decoding' but not 'structural encoding'. The enhanced cortical response to perception of moderately intense sad facial expressions following citalopram administration may relate to the cognitive processing of the social relevance or significance of such ambiguous stimuli.


Vascular risk profiles for dementia and Alzheimer's disease in very old people: a population-based longitudinal study.
Numerous studies have linked individual vascular factors to dementia including Alzheimer's disease (AD). We investigated different vascular risk profiles in relation to dementia and AD among very old people. A standardized follow-up procedure was applied three times to a dementia-free cohort (n=1270, age >or= 75) over a nine-year period to detect dementia and AD cases using the DSM-III-R criteria. We examined two vascular risk profiles, which were scored by counting the number of corresponding vascular factors: 1) atherosclerotic profile included systolic pressure >or= 160 mmHg, diabetes/prediabetes, and stroke; and 2) cerebral hypoperfusion profile constituted diastolic pressure < 70 mmHg, pulse pressure < 70 mmHg, and heart failure. Data were analyzed with Cox proportional-hazards models controlling for major potential confounders. During the 6406 person-years of follow-up, 428 subjects developed dementia, including 328 AD cases. All components of vascular profiles were significantly or marginally associated with increased dementia risk. The risk of dementias was increased with increasing score of both risk profiles (p for trend <or= 0.001); subjects with a score >or= 2 in either profile had an approximately twofold-increased risk for dementia and AD. These data suggest that aggregation of atherosclerotic- and hypoperfusion-related vascular factors increases the risk of dementia in very old people. Severe cerebral atherosclerosis and insufficient perfusion are involved in the development of dementia including AD.


[Osteoarthritis - which approach is evidence based?].
Paracetamol is the first choice mediation for osteoarthritis. The analgetic potential of NSAIDs is slightly higher and they also have some antiphlogistic effect, but their use has to be strictly limited to a short period of time. They should mainly be used in the therapy of the acute and painful phase of osteoarthritis. Among the NSAIDs, Diclofenac is the medication of first choice. In patients with an increased risk of gastrointestinal complications, a protonpumpinhibitor should be added. Patients with cardiovascular risk factors should receive NSAIDs only in case of no appropriate alternative treatments. Opioids have their place in osteoarthritis treatment and should be part of an individualized pain regime, which should also contain a pain diary and proactive monitoring. It is important to emphasize the positive effects of physical activity on the function of the joints as well as the negative effect of overweight and immobility.


"Aspirinated" platelets are hemostatic in thrombocytopenic rats with "nonaspirinated" vessel walls--evidence from an exchange transfusion model.
The contrasting effects of aspirin on bleeding time (BT) might be related to the drug's inhibitory activity on platelets and vascular prostaglandin I2 (PGI2). To test this, we developed an exchange transfusion model in the rat and studied the BT in animals whose platelets but not vessels had been exposed to aspirin. Rats with severe experimental thrombocytopenia were exchange-transfused with blood from normocythemic rats pretreated with aspirin 6 hr before. The platelet count was raised from 2% to about 70% of basal level and the BT returned to control values even though the platelets neither responded to arachidonic acid nor produced detectable amounts of malondialdehyde and vascular PGI2 was not inhibited. These results indicate that "aspirinated" platelets may be hemostatically active and that the BT is not necessarily affected by unbalanced prostaglandin production in platelets and the vessell wall.


Predicting Pathologic Response of Esophageal Cancer to Neoadjuvant Chemotherapy: The Implications of Metabolic Nodal Response for Personalized Therapy.
Only a minority of esophageal cancers demonstrates a pathologic tumor response (pTR) to neoadjuvant chemotherapy (NAC). 18F-FDG PET/CT is often used for restaging after NAC and to assess response. Increasingly, it is used during therapy to identify unresponsive tumors and predict pTR, using avidity of the primary tumor alone. However, definitions of such metabolic tumor response (mTR) vary. We aimed to comprehensively reevaluate metabolic response assessment using accepted parameters, as well as novel concepts of metabolic nodal stage (mN) and metabolic nodal response (mNR). This was a single-center retrospective U.K. cohort study. All patients with esophageal cancer staged before NAC with PET/CT and after with CT or PET/CT and undergoing resection from 2006 to 2014 were identified. pTR was defined as Mandard tumor regression grade 1-3; imaging parameters included metrics of tumor avidity (SUVmax/mean/peak), composites of avidity and volume (including metabolic tumor volume), nodal SUVmax, and our new concepts of mN stage and mNR. Eighty-two (27.2%) of 301 patients demonstrated pTR. No pre-NAC PET parameters predicted pTR. In 220 patients restaged by PET/CT, the optimal tumor ΔSUVmax threshold was a 77.8% reduction. This was as sensitive as the current PERCIST 30% reduction, but more specific with a higher negative predictive value (P < 0.001). ΔSUVmax and Δlength independently predicted pTR, and composite avidity/spatial metrics outperformed avidity alone. Although both mTR and mNR were associated with pTR, in 82 patients with 18F-FDG-avid nodes before NAC we observed mNR in 10 (12.2%) not demonstrating mTR. Current definitions of metabolic response are suboptimal and too simplistic. Composite avidity/volume measures improve prediction. mNR may further improve response assessment, by specifically assessing metastatic tumor subpopulations, likely responsible for disease relapse, and should be urgently assessed when considering aborting therapy on the basis of mTR alone.


Acute endothelial swelling is induced in endoneurial microvessels by ischemia.
Structural alterations of endoneurial microvessels occur in diabetic neuropathy and are statistically associated with severity of nerve fiber loss and teased fiber abnormality. It is therefore hypothesized that the microvessel alterations may cause or contribute to pathologic alterations of nerve fibers in diabetic neuropathy, possibly through hypoxic injury. The mechanism of the microvessel change in diabetic neuropathy is unknown. The role of microvessels and details of microvessel structure in other possible ischemic neuropathies has not been studied completely. Already there is evidence that hypoxia induces endothelial swelling but this has not been characterized or quantitated in nerve. To determine the acute morphologic effect of ischemia on ultrastructural features of transverse profiles of endoneurial microvessels major pelvic arteries were ligated in rats. At 36 h mean lumen and mural areas were greater in ischemic than in control nerves. All components (endothelium, pericytes and basement membrane) were on average greater in ischemic than controls. The greatest increase was in endothelial cells. In these cells swollen mitochondria were abundant. This study demonstrates that acute ischemia induces swelling of the cells and organelles of endoneurial microvessels.


Acute effects on human sperm exposed in vitro to cadmium chloride and diisobutyl phthalate.
Epidemiological studies reported a negative relationship between concentrations of heavy metals and phthalates in seminal fluid and semen quality, likely compromising male fertility potential. The aim of this study was to investigate the in vitro effects of cadmium chloride (CdCl2), a common heavy metal, and diisobutyl phthalate (DIBP), a common phthalate ester, on human sperm functions necessary for fertilization. After in vitro incubation of spermatozoa with 10 µM CdCl2 or 100 and 200 µM DIBP for 24 h, a significant decrease of sperm progressive and hyperactivated motility was observed. The exposure to each of the two toxic agents also induced spontaneous sperm acrosome reaction and blunted the physiological response to progesterone. Both agents induced an increase of caspase activity suggesting triggering of an apoptotic pathway. Our results suggest that acute exposure of spermatozoa to these pollutants may impair sperm ability to reach and fertilize the oocyte.


Differences in Cognitive Function in Women and Men with Diabetic Peripheral Neuropathy with or without Pain.
The aim of this study was to analyse the differences in cognitive function between women and men with type-2 diabetes mellitus (DMT2) and diabetic peripheral neuropathy (DPN) with and without diabetic neuropathic pain (DNP), and the factors associated with cognitive function in each sex. A cross-sectional study of 149 patients with DMT2 and DPN was performed. Sociodemographic and clinical variables, Test Your Memory (TYM) for cognitive assessment, anxiety and depression (HADS), quality of life (SF-12v2) and sleep characteristics (MOS-sleep) were measured. A high percentage of women presented cognitive impairment (50% vs. 36.1%) and they scored lower on the TYM (mean = 40.77; SD = 6.03 vs. mean = 42.49; SD = 6.05). Women with DNP scored lower on calculation tasks (3.17 vs. 3.52) than men with DNP, while women without DNP scored lower on retrograde memory (2.70 vs. 3.74), executive function (3.83 vs. 4.25) and similarities (2.51 vs. 3.12) than men without DNP. Being older (B = -0.181) and presenting cardiovascular risk factors (B = -5.059) were associated with worse cognitive function in women, while in men this was associated with older age (B = -0.154), a longer duration of diabetes (B = -0.319) and the presence of depression (B = -0.363). Women with and without DNP obtained worse results in cognitive function. However, the presence of pain had a greater impact on the different dimensions in men.


Guideline compliance for bridging anticoagulation use in vitamin-K antagonist patients; practice variation and factors associated with non-compliance.
Bridging anticoagulation is used in vitamin-K antagonist (VKA) patients undergoing invasive procedures and involves complex risk assessment in order to prevent thromboembolic and bleeding outcomes. Our aim was to assess guideline compliance and identify factors associated with bridging and especially, non-compliant bridging. A retrospective review of 256 patient records in 13 Dutch hospitals was performed. Demographic, clinical, surgical and care delivery characteristics were collected. Compliance to the American College of Chest Physicians ninth edition guideline (AT9) was assessed. Multilevel regression models were built to explain bridging use and predict non-compliance. Bridging use varied from 15.0 to 83.3% (mean = 41.8%) of patients per hospital, whereas guideline compliance varied from 20.0 to 88.2% (mean = 68.5%) per hospital. Both established thromboembolic risk factors and characteristics outside thromboembolic risk assessment were associated with bridging use. Predictors for overuse were gastrointestinal surgery (OR 14.85, 95% CI 2.69-81.99), vascular surgery (OR 13.01, 95% CI 1.83-92.30), non-elective surgery (OR 8.67, 95% CI 1.67-45.14), lowest 25th percentile socioeconomic status (OR 0.33, 95% CI 0.11-1.02) and use of VKA reversal agents (OR 0.22, 95% CI 0.04-1.16). Bridging anticoagulation practice was not compliant with the AT9 in 31.5% of patients. The aggregated AT9 thromboembolic risk was inferior to individual thromboembolic risk factors and other characteristics in explaining bridging use. Therefor the AT9 risk seems less important for the decision making in everyday practice. Additionally, a heterogeneous implementation of the guideline between hospitals was found. Further research and interventions are needed to improve bridging anticoagulation practice in VKA patients.


Catalytic, asymmetric alpha-chlorination of acid halides.
We present a full account of a tandem catalytic, asymmetric chlorination/esterification process that produces highly optically enriched alpha-chloroesters from inexpensive, commercially available acid halides using cinchona alkaloid derivatives as catalysts and polychlorinated quinones as halogenating agents. We have performed kinetics and control experiments to investigate the reaction mechanism and establish conditions under which the reactions can be best performed. We have developed NaH and NaHCO3 shuttle base systems as the easiest and most cost-effective ways of conducting the reactions, rendering the methodology economically competitive with known chiral halogenation procedures. We have also demonstrated the utility of our reactions by converting the products to synthetically useful derivatives.


Ion-Type Dependence of DNA Electronic Excitation in Water under Proton, α-Particle, and Carbon Ion Irradiation: A First-Principles Simulation Study.
Understanding how the electronic excitation of DNA changes in response to different high-energy particles is central to advancing ion beam cancer therapy and other related approaches, such as boron neutron capture therapy. While protons have been the predominant ions of choice in ion beam cancer therapy, heavier ions, particularly carbon ions, have drawn significant attention over the past decade. Carbon ions are expected to transfer larger amounts of energy according to linear response theory. However, molecular-level details of the electronic excitation under heavier ion irradiation remain unknown. In this work, we use real-time time-dependent density functional theory simulations to examine the quantum-mechanical details of DNA electronic excitations in water under proton, α-particle, and carbon ion irradiation. Our results show that the energy transfer does indeed increase for the heavier ions, while the excitation remains highly conformal. However, the increase in the energy transfer rate, measured by electronic stopping power, does not match the prediction by the linear response model, even when accounting for the velocity dependence of the irradiating ion's charge. The simulations also reveal that while the number of holes generated on DNA increases for heavier ions, the increase is only partially responsible for the larger stopping power. Larger numbers of highly energetic holes formed from the heavier ions also contribute significantly to the increased electronic stopping power.


Electroconvulsive therapy and complaints of memory dysfunction: a prospective three-year follow-up study.
Self-reports of memory problems have been evaluated prospectively in depressed patients receiving bilateral ECT or unilateral ECT, and in depressed patients receiving treatments other than ECT. Depressed patients did not complain of poor memory at seven months after hospitalization. Compared to bilateral ECT, right unilateral ECT was associated with only mild memory complaints. At three years after treatment approximately one-half of the persons who had received bilateral ECT reported poor memory. These reports seemed to be influenced by three factors: (1) recurrence or persistence of conditions that were present before ECT; (2) the experience of amnesia initially associated with ECT and a subsequent tendency to question if memory had ever recovered; and (3) impaired memory for events that had occurred up to six months before treatment and up to about two months afterwards.


[Systemic lupus erythematosus with steroid induced non-alcoholic steatohepatitis: a case report].
A 56-year old Japanese female was admitted to our hospital because of the increased levels of serum AST, ALT, and gamma-GTP. She was diagnosed with systemic lupus erythematosus in September, 1996 and had been on a regular glucocorticoid therapy since then. Abdominal ultrasonography showed the mild fatty liver, and hepatic histopathology revealed a typical and remarkable steatohepatitis, a remarkable neutrophil infiltration, and Mallory bodies. Because she had no history of alcohol-drinking, diagnosis of non-alcoholic steatohepatitis (NASH) was made. Treatment was started with a low-calorie diet, bed-rest, and an oral administration of alpha-tocopherol and bezafibrate with favorable effects on her serum levels of AST, ALT, gamma-GTP, and LDH. When a patient on a glucocorticoid therapy shows signs of fatty liver, diabetes mellitus, hyperlipidemia, an insulin resistance, NASH should be considered as one of the differential diagnosis. This is particularly important since proper therapy with a low-calorie diet and drugs with anti-oxidant activities improve this potentially progressive disease before resulting in liver cirrhosis and hepatic carcinoma.


Role of the actin cytoskeleton in insulin action.
Insulin has diverse effects on cells, including stimulation of glucose transport, gene expression, and alterations of cell morphology. The hormone mediates these effects by activation of signaling pathways which utilize, 1) adaptor molecules such as the insulin receptor substrates (IRS), the Src and collagen homologs (Shc), and the growth factor receptor binding protein 2 (Grb2); 2) lipid kinases such as phosphatidylinositol 3-kinase (PI 3-Kinase); 3) small G proteins; and 4) serine, threonine, and tyrosine kinases. The activation of such signaling molecules by insulin is now well established, but we do not yet fully understand the mechanisms integrating these seemingly diverse pathways. Here, we discuss the involvement of the actin cytoskeleton in the propagation and regulation of insulin signals. In muscle cells in culture, insulin induces a rapid actin filament reorganization that coincides with plasma membrane ruffling and intense accumulation of pinocytotic vesicles. Initiation of these effects of insulin requires an intact actin cytoskeleton and activation of PI 3-kinase. We observed recruitment PI 3-kinase subunits and glucose transporter proteins to regions of reorganized actin. In both muscle and adipose cells, actin disassembly inhibited early insulin-induced events such as recruitment of glucose transporters to the cell surface and enhanced glucose transport. Additionally, actin disassembly inhibited more prolonged effects of insulin, including DNA synthesis and expression of immediate early genes such as c-fos. Intact actin filaments appear to be essential for mediation of early events such as association of Shc with Grb2 in response to insulin, which leads to stimulation of gene expression. Preliminary observations support a role for focal adhesion signaling complexes in insulin action. These observations suggest that the actin cytoskeleton facilitates propagation of the morphological, metabolic, and nuclear effects of insulin by regulating proper subcellular distribution of signaling molecules that participate in the insulin signaling pathway.


The effect of oxidative stress on the mutation rate of Mycobacterium tuberculosis with impaired catalase/peroxidase function.
To determine the effect of oxidative stress on isoniazid-resistant Mycobacterium tuberculosis deficient in catalase/peroxidase activity to varying degrees through mutation in katG. The mutation rate was determined for a set of isogenic strains with different katG alleles giving different catalase and/or peroxidase activities following exposure to the oxidizing agent, hydrogen peroxide. Mutants were selected on rifampicin, and the location and nature of the mutation were identified by sequencing the rpoB gene. No evidence was found to suggest that strains that had impaired catalase/peroxidase activity were hypermutable, and the presence of excess hydrogen peroxide had no effect on the mutation rate. An unusual pattern of mutations in rpoB was observed in catalase-deficient strains with only 3 of 66 having mutations within the rifampicin resistance-determining region. The mutation rate of M. tuberculosis in response to oxidative stress is not increased in strains with significant deficits in catalase and peroxidase activity. Our data suggest that isoniazid-resistant strains compensate for their reduced ability to detoxify oxidative stress effectively. Interestingly, mutations were found in unusual locations at positions similar to those found in clinical isoniazid-resistant strains.


The Transcriptional and Epigenetic Landscape of Cancer Cell Lineage Plasticity.
Lineage plasticity, a process whereby cells change their phenotype to take on a different molecular and/or histologic identity, is a key driver of cancer progression and therapy resistance. Although underlying genetic changes within the tumor can enhance lineage plasticity, it is predominantly a dynamic process controlled by transcriptional and epigenetic dysregulation. This review explores the transcriptional and epigenetic regulators of lineage plasticity and their interplay with other features of malignancy, such as dysregulated metabolism, the tumor microenvironment, and immune evasion. We also discuss strategies for the detection and treatment of highly plastic tumors. Lineage plasticity is a hallmark of cancer and a critical facilitator of other oncogenic features such as metastasis, therapy resistance, dysregulated metabolism, and immune evasion. It is essential that the molecular mechanisms of lineage plasticity are elucidated to enable the development of strategies to effectively target this phenomenon. In this review, we describe key transcriptional and epigenetic regulators of cancer cell plasticity, in the process highlighting therapeutic approaches that may be harnessed for patient benefit.


TOPAS Simulation of the Mevion S250 compact proton therapy unit.
As proton therapy becomes increasingly popular, so does the need for Monte Carlo simulation studies involving accurate beam line modeling of proton treatment units. In this study, the 24 beam configurations of the Mevion S250 proton therapy system installed recently at our institution were modeled using the TOolkit for PArticle Simulation (TOPAS) code. Pristine Bragg peak, spread out Bragg peak (SOBP), and lateral beam profile dose distributions were simulated and matched to the measurements taken during commissioning of the unit. Differences in the range for all Percent Depth Dose (PDD) curves between measured and simulated data agreed to within 0.1 cm. For SOBP scans, the SOBP widths all agreed to within 0.3 cm. With regards to lateral beam profile comparisons between the measured and simulated data, the penumbras differed by less than 1 mm and the flatness differed by less than 1% in nearly all cases. This study shows that Monte Carlo simulation studies involving the Mevion S250 proton therapy unit can be a viable tool in commissioning and verification of the proton treatment planning system.


Modelling spatiotemporal dynamics of cerebral blood flow using multiple-timepoint arterial spin labelling MRI.
Introduction: Cerebral blood flow (CBF) is an important physiological parameter that can be quantified non-invasively using arterial spin labelling (ASL) imaging. Although most ASL studies are based on single-timepoint strategies, multi-timepoint approaches (multiple-PLD) in combination with appropriate model fitting strategies may be beneficial not only to improve CBF quantification but also to retrieve other physiological information of interest. Methods: In this work, we tested several kinetic models for the fitting of multiple-PLD pCASL data in a group of 10 healthy subjects. In particular, we extended the standard kinetic model by incorporating dispersion effects and the macrovascular contribution and assessed their individual and combined effect on CBF quantification. These assessments were performed using two pseudo-continuous ASL (pCASL) datasets acquired in the same subjects but during two conditions mimicking different CBF dynamics: normocapnia and hypercapnia (achieved through a CO2 stimulus). Results: All kinetic models quantified and highlighted the different CBF spatiotemporal dynamics between the two conditions. Hypercapnia led to an increase in CBF whilst decreasing arterial transit time (ATT) and arterial blood volume (aBV). When comparing the different kinetic models, the incorporation of dispersion effects yielded a significant decrease in CBF (∼10-22%) and ATT (∼17-26%), whilst aBV (∼44-74%) increased, and this was observed in both conditions. The extended model that includes dispersion effects and the macrovascular component has been shown to provide the best fit to both datasets. Conclusion: Our results support the use of extended models that include the macrovascular component and dispersion effects when modelling multiple-PLD pCASL data.


Downregulation of connexin40 is associated with coronary endothelial cell dysfunction in streptozotocin-induced diabetic mice.
Vascular endothelial cells (ECs) play a major role in regulating vascular tone and in revascularization. There is increasing evidence showing endothelial dysfunction in diabetes, although little is known about the contribution of connexins (Cxs) to vascular complications in the diabetic heart. This study was designed to investigate the role of Cxs in coronary endothelial dysfunction in diabetic mice. Coronary ECs isolated from diabetic mice exhibit lowered protein levels of Cx37 and Cx40 (but not Cx43) and a loss of gap junction intercellular communication (GJIC). Vasodilatation induced by the assumed contribution of EC-dependent hyperpolarization was significantly reduced in the diabetic coronary artery (CA). Cx40-specific inhibitory peptide (40)GAP27 strongly attenuated endothelium-dependent relaxation in diabetic CA at the concentration that does not affect the relaxation in control CA, suggesting that the total amount of Cx40 is lower in diabetic CA than in control CA. In diabetic mice, coronary capillary density was significantly decreased in vivo. In vitro, GJIC inhibitor attenuated the ability of EC capillary network formation. High-glucose treatment caused a decrease in Cx40 protein expression in ECs and impaired endothelial capillary network formation, which was restored by Cx40 overexpression. Furthermore, we found that the hyperglycemia-induced decrease in Cx40 was associated with inhibited protein expression of Sp1, a transcriptional factor that regulates Cx40 expression. These data suggest that downregulation of Cx40 protein expression and resultant inhibition of GJIC contribute to coronary vascular dysfunction in diabetes.


Hormonal status modulates circulating endothelial progenitor cells.
Endothelial progenitor cells (EPCs) may have an important role in vascular homeostasis and repair. We examined the level of circulating EPCs in pre- (n = 22; mean age 28.7 years), and postmenopausal healthy females without (n = 30; mean age 61.6 years) or under current hormone replacement therapy (HRT) (n = 19; mean age 59.8 years). Premenopausal females had the highest level of circulating EPCs (0.147 +/- 0.076 per thousand of polymorphnuclear cells). The level of EPCs was lowest in postmenopausal females (0.094 +/- 0.058 per thousand), and increased significantly with HRT on average by 25.5%. In addition, the proliferative capacity of circulating EPCs was assessed under cell culture conditions. This capacity was significantly increased in EPCs isolated from postmenopausal subjects under current HRT as compared to corresponding samples obtained from postmenopausal females without HRT. This observation is in line with the hypothesis that the hormonal status in females modulates the cardiovascular risk and that circulating EPCs could be involved in this phenomenon.