RNA from an immediate early region of the type 1 herpes simplex virus genome is present in the trigeminal ganglia of latently infected mice.
Transcription of the type 1 herpes simplex virus (HSV-1) genome in trigeminal ganglia of latently infected mice was studied using in situ hybridization. Probes representative of each temporal gene class were used to determine the regions of the genome that encode the transcripts present in latently infected cells. Probes encoding HSV-1 sequences of the five immediate early genes and representative early (thymidine kinase), early-late (major capsid protein), and late (glycoprotein C) genes were used in these experiments. Of the probes tested, only those encoding the immediate early gene product infected-cell polypeptide (ICP) 0 hybridized to RNA in latently infected tissues. Probes containing the other immediate early genes (ICP4, ICP22, ICP27, and ICP47) and the representative early, early-late, and late genes did not hybridize. Two probes covering approximately equal to 30% of the HSV-1 genome and encoding over 20 early and late transcripts also did not hybridize to RNA in latently infected tissues. These results, with probes spanning greater than 60% of the HSV-1 genome, suggest that transcription of the HSV-1 genome is restricted to one region in latently infected mouse trigeminal ganglia.


Sex chromosomes and the evolution of sexual dimorphism: lessons from the genome.
Females and males of many animals exhibit a striking array of sexual dimorphisms, ranging from the primary differences of the gametes and gonads to the somatic differences often seen in behavior, morphology, and physiology. These differences raise many questions regarding how such divergent phenotypes can arise from a genome that is largely shared between the sexes. Recent progress in genomics has revealed some of the actual genetic mechanisms that create separate sex-specific phenotypes, and the evidence indicates that thousands of genes across all portions of the genome contribute to male and female forms through sex-biased gene expression. Related work has begun to define the strength and influence of sex-specific evolutionary forces that shape these phenotypic dimorphisms and how they in turn affect the genome. Additionally, theory has long suggested that the evolution of sexual dimorphism is facilitated by sex chromosomes, as these are the only portions of the genome that differ between males and females. Genomic analysis indicates that there is indeed a relationship between sexual dimorphism and the sex chromosomes. However, the connection is far more complicated than current theory allows, and this may ultimately require a reexamination of the assumptions so that predictions match the accumulating empirical data.


Endoscopically Assisted Middle Cranial Fossa Reconstruction Following Traumatic Intracranial Intrusion of the Mandibular Condyle.
Traumatic intracranial intrusions of the mandibular condyle are a rare injury pattern that carries significant morbid outcomes due to potential for neurological, otological, temporomandibular joint damage, or joint ankylosis. Treatment modalities involve either closed reduction, open approaches via transcranial or subtemporal approaches, or delayed total joint replacement. This paper presents a clinical report of a traumatic intracranial intrusion of the mandibular condyle treated with an endoscopically assisted preauricular approach to reconstruct the middle cranial fossa and temporomandibular articular disc. Endoscopically assisted techniques allow for minimally invasive and less morbid surgical interventions to this pathology, with greater precision than closed reduction techniques.


Therapeutic efficacy of folate receptor-targeted amphiphilic cyclodextrin nanoparticles as a novel vehicle for paclitaxel delivery in breast cancer.
The aim of this study is to test folate-conjugated cyclodextrin nanoparticles (FCD-1 and FCD-2) as a vehicle for reducing toxicity and increasing the antitumor efficacy of paclitaxel especially for metastatic breast cancer. For the evaluation of PCX-loaded FCD nanoparticles, animal studies were realised in terms of survival rate, tumour size, weight change, metastazis and histopathological examination. FCD-1 displayed significant advantages such as efficient targeting of folate receptor positive breast cancer cells and having considerably lower toxicity compared to that of Cremophor®. When loaded with paclitaxel, FCD-1 nanoparticles, which have smaller particle size, neutral zeta potential, high encapsulation efficiency and better loading capacity for controlled release, emerged as an effective formulation in terms of cytotoxicity and high cellular uptake. In an experimental breast cancer model, anticancer activity of these nanoparticles were compatible with that of paclitaxel in Cremophor® however repeated administrations of FCD-1 nanoparticles were better tolerated by the animals. These nanoparticles were able to localise in tumour site. Both paclitaxel-loaded FCD-1 and FCD-2 significantly reduced tumour burden while FCD-1 significantly improved the survival. Folate-conjugated amphiphilic cyclodextrin nanoparticles can be considered as promising Cremophor®-free, low-toxicity and efficient active drug delivery systems for paclitaxel.


Elbasvir plus grazoprevir in patients with hepatitis C virus infection and stage 4-5 chronic kidney disease: clinical, virological, and health-related quality-of-life outcomes from a phase 3, multicentre, randomised, double-blind, placebo-controlled trial.
In the C-SURFER study, therapy with the all-oral elbasvir plus grazoprevir regimen for 12 weeks in patients with chronic hepatitis C virus (HCV) infection and stage 4-5 chronic kidney disease resulted in a high rate of virological cure compared with placebo. Here, we report sustained virological response (SVR), safety data, health-related quality-of-life (HRQOL), and virological resistance analyses in patients in C-SURFER who received immediate antiviral therapy or who received placebo before therapy. In this phase 3, multicentre, randomised, placebo-controlled study, we randomly assigned adults with HCV genotype 1 infection and stage 4-5 chronic kidney disease enrolled at 68 centres worldwide to either elbasvir 50 mg plus grazoprevir 100 mg once per day for 12 weeks (immediate treatment group) or placebo for 12 weeks followed by elbasvir 50 mg plus grazoprevir 100 mg once per day for 12 weeks beginning at week 16 (deferred treatment group). The primary safety and efficacy endpoints for the immediate treatment group and placebo phase of the deferred treatment group have been reported previously. Here, we report safety and efficacy data for the treatment phase of the deferred treatment group, as well as HRQOL assessed using the 36-Item Short Form Health Survey for all groups, and baseline and treatment-emergent resistance-associated substitutions (RASs). SVR at 12 weeks (SVR12) was assessed in the modified full analysis set (FAS), defined as all patients excluding those who did not receive at least one dose of study drug, who died, or who discontinued the study before the end of treatment for reasons determined to be unrelated to HCV treatment. This trial is registered with ClinicalTrials.gov, Number NCT02092350. Between March 30 and Nov 28, 2014, 235 patients were enrolled and received at least one dose of study drug. The modified FAS included 116 patients assigned to immediate treatment and 99 assigned to deferred treatment. 115 (99·1%; 95% CI 95·3-100·0) of 116 assigned to immediate treatment achieved SVR12 compared with 97 (98·0%; 92·9-99·7) of 99 assigned to deferred treatment. In patients with genotype 1a infections, SVR12 was achieved by 11 (84·6%) of 13 patients with detectable baseline NS5A RASs and in 98 (100%) of 98 without. HRQOL did not differ at week 12 between immediate treatment and the placebo phase of deferred treatment. Safety was generally similar between patients receiving immediate treatment and those receiving placebo in the deferred treatment group. One serious adverse event during deferred treatment (interstitial nephritis) and one during the placebo phase of deferred treatment (raised lipase concentration) were deemed related to study drug. Four patients died, one who received immediate treatment (cardiac arrest) and three who received deferred treatment (aortic aneurysm, pneumonia, and unknown cause); all four deaths were considered unrelated to study drugs. Of the three deaths in the deferred treatment group, one occurred during placebo treatment and two occurred before starting active treatment. There were no notable differences in aminotransferase elevations in the deferred treatment group compared with the immediate treatment group, and no patients in the deferred treatment group had total bilirubin elevations. These data add to the growing body of clinical evidence for the fixed-dose combination regimen of elbasvir plus grazoprevir for 12 weeks and support use of this therapy in patients with HCV genotype 1 infection and stage 4-5 chronic kidney disease. Merck Sharp & Dohme.


[Expression of survivin and its clinical significance in non-small cell lung cancer].
To investigate the expression of survivin in non-small cell lung cancer (NSCLC) and analyze its relationship with clinico pathological features and the expression of p53. The expression of survivin and p53 in 61 cases of NSCLC were detected by immunohistochemistry using monoclonal antibodies against human survivin protein and P53, respectively. The expression of survivin in NSCLC could be detected in cytoplasm and/or in nucleus. The overall immunoreactivity was found in 77.0% (47/61) of tumors. Cytoplasmic, nuclear and both immunoreactivities were present in 26.2% (16/61), 8.2% (5/61) and 42.62% (26/61) respectively. The cytoplasmic and nuclear immunoreactivities in squamous cell carcinoma (57.6%, 19/33) were higher than that in adenocarcinoma (25%, 7/28) and there was significant difference (P<0.05). The positive expression rates of survivin in P53 positive and negative were 54.3% (19/35) and 26.9%(7/26) respectively and there was significant difference (P<0.05). Survivin expression levels did not correlate with the patient's gender, age, clinical stage, histological differentiation, lymph node metastasis status or survival. survivin was overexpressed in NSCLC. It correlated with pathological type that mainly showed the positive expression rate in squamous cell carcinoma was higher than that in adenocarcinoma. It may play an important role in the carcinogenesis and development of NSCLC by inhibiting tumor cell apoptosis and facilitating angiogenesis. Inactivation of antioncogene p53 and expression of survivin may play synergetic roles in carcinogenesis of NSCLC.


[Improvement of angina, quality of life, and working capacity after enhanced external counterpulsation].
This case report describes a 43 year-old woman who had had 15 hospital admissions in six years due to chest pain. Repeated coronary angiography showed no significant stenoses, but myocardial scintigraphy revealed reversible ischaemia. In spite of optimal medical treatment, she was in Canadian Cardiovascular Society (CCS) class 3 with a poor quality of life (QoL). After a course of enhanced external counterpulsation the CCS class was reduced to 1, and QoL was significantly improved and remained high for the following three years, resulting in only one hospital admission due to chest pain. The case illustrates that EECP is an efficient therapy for angina pectoris in patients, who are not amenable for invasive procedures.


Elongated Nanoparticle Aggregates in Cancer Cells for Mechanical Destruction with Low Frequency Rotating Magnetic Field.
Magnetic nanoparticles (MNPs) functionalized with targeting moieties can recognize specific cell components and induce mechanical actuation under magnetic field. Their size is adequate for reaching tumors and targeting cancer cells. However, due to the nanometric size, the force generated by MNPs is smaller than the force required for largely disrupting key components of cells. Here, we show the magnetic assembly process of the nanoparticles inside the cells, to form elongated aggregates with the size required to produce elevated mechanical forces. We synthesized iron oxide nanoparticles doped with zinc, to obtain high magnetization, and functionalized with the epidermal growth factor (EGF) peptide for targeting cancer cells. Under a low frequency rotating magnetic field at 15 Hz and 40 mT, the internalized EGF-MNPs formed elongated aggregates and generated hundreds of pN to dramatically damage the plasma and lysosomal membranes. The physical disruption, including leakage of lysosomal hydrolases into the cytosol, led to programmed cell death and necrosis. Our work provides a novel strategy of designing magnetic nanomedicines for mechanical destruction of cancer cells.


Normal paraspinal muscle electromyographic fatigue characteristics in patients with primary fibromyalgia.
Paraspinal muscle fatigue mechanisms were compared in 14 primary fibromyalgia patients and 14 age and sex matched normal subjects using a standardized 60-s isometric endurance test of the paraspinal muscles, during which surface integrated electromyographic (IEMG) activity was recorded. Fatigue-induced IEMG increases were similar for both groups during the initial 40 s (up to 112 +/- 20% and 111 +/- 6% of initial values in patients and normal subjects respectively). Thereafter, IEMG fell significantly in patients (P < 0.05) but only slightly in controls, so that at 58 s IEMG was 102 +/- 13% in patients and 109 +/- 12% in controls. If patients were divided according to body mass index (BMI, range 19-25 in controls) those with a BMI < 26 (n = 5) showed IEMG changes similar to those of control subjects throughout the test, while obese patients with BMI > 26 (n = 9) showed greater IEMG declines after 40 s than either normal subjects or in the fibromyalgia group as a whole. Paraspinal muscle fatigue mechanisms appear normal in primary fibromyalgia patients. Isometric force maintenance in overweight patients, despite IEMG declines, illustrates the action of intrinsic fatigue resistance mechanisms which were presumably utilized to a greater extent in these patients to cope with the extra load.


The long non-coding RNA Gm10768 activates hepatic gluconeogenesis by sequestering microRNA-214 in mice.
Overactivated hepatic gluconeogenesis contributes to the pathogenesis of metabolic disorders, including type 2 diabetes. Precise control of hepatic gluconeogenesis is thus critical for maintaining whole-body metabolic homeostasis. Long non-coding RNAs (lncRNAs) have been shown to play key roles in diseases by regulating diverse biological processes, but the function of lncRNAs in maintaining normal physiology, particularly glucose homeostasis in the liver, remains largely unexplored. We identified a novel liver-enriched long non-coding RNA, Gm10768, and examined its expression patterns under pathophysiological conditions. We further adopted gain- and loss-of-function strategies to explore the effect of Gm10768 on hepatic glucose metabolism and the possible molecular mechanism involved. Our results showed that the expression of Gm10768 was significantly increased in the liver of fasted mice and was induced by gluconeogenic hormonal stimuli. Functionally, overexpression of Gm10768 activated hepatic gluconeogenesis in a cell-autonomous manner. In contrast, depletion of Gm10768 suppressed hepatic glucose production both in vitro and in vivo Adenovirus-mediated hepatic knockdown of Gm10768 improved glucose tolerance and hyperglycemia of diabetic db/db mice. Mechanistically, Gm10768 sequestrated microRNA-214 (miR-214) to relieve its suppression on activating transcription factor 4 (ATF4), a positive regulator of hepatic gluconeogenesis. Taken together, we identified Gm10768 as a new lncRNA activating hepatic gluconeogenesis through antagonizing miR-214 in mice.


Development of morphine glucuronidation in premature neonates.
In premature neonates (25-34 weeks gestation) who were given morphine intravenously during the first 24 h of life, only morphine, and morphine-3-glucuronide (M3G) were detected in plasma obtained after a 2-hour loading infusion, but morphine-6-glucuronide (M6G) could also be quantified following 24 h of continuous infusion. M3G/morphine and M6G/morphine plasma concentration ratios increased significantly with increasing birth weight. However, the M6G/M3G plasma concentration ratio decreased with increasing birth weight (and gestational age), thus providing the first indication in vivo of the differential development of uridinediphosphate glucuronosyltransferases in humans.


The rabbit kappa 1 b5 immunoglobulin gene: another J region gene cluster with only one functional J gene segment?
Our previous analysis of an immunoglobulin gene encoding the rabbit kappa chain of b4 allotype revealed that of the five J-like sequences in the J kappa cluster of this gene, only one, J2, appeared to be functional. This unusual ratio of J pseudogenes to functional J genes is unique among all J clusters of light and heavy chain genes of all species examined to date, including the cluster from the rabbit kappa 2 isotype, and must have consequences for diversity generation of b4 immunoglobulins. The fact that the only two known b5 J kappa sequences are different from the functional J2 of the b4 allotype prompted investigation of the b5 J kappa cluster to determine whether it resembled the b4 cluster, or the more typical mouse or human J kappa clusters. Our analysis of the b5 gene reveals a J kappa cluster strikingly similar to that of b4; apparent defects occur in all J sequences except J2. Although J2 is apparently functional, it differs from the J2 of the b4 locus by four nucleotide and three amino acid substitutions. The unusually high degree of sequence similarity previously observed between the b4 and b5 loci in the noncoding (vs the coding) regions extends through the newly sequenced DNA segment and remains an enigma.


Adiabatic thermo-optic Mach-Zehnder switch.
In this Letter, we propose and demonstrate a high-speed and power-efficient thermo-optic switch using an adiabatic bend with a directly integrated silicon heater to minimize the heat capacity and therein maximize the performance of the thermo-optic switch. A rapid, τ=2.4 μs thermal time constant and a low electrical power consumption of P(π)=12.7 mW/π-phase shift were demonstrated representing a P(π)τ product of only 30.5 mW·μs in a compact device with a phase shifter of only ~10 μm long.


Mechanisms of action of selenium and vitamin E in protection of biological membranes.
Opinions vary as to the mode of action of vitamin E and selenium. Some argue that they act as nonspecific biological antioxidants. Others propose that the functions of the two substances are distinct and that vitamin E acts as a true vitamin in addition to functioning as a lipid-soluble antioxidant. Support for the "Biological Antioxidant Theory" is largely circumstantial. However, lipoperoxides have been detected in adipose tissues of vitamin E-deficient animals, and increased rates of in vitro peroxidation have been demonstrated in homogenates of several tissues of selenium and vitamin E-deficient animals. The basis of the antioxygenic role of selenium in these systems was elucidated by the discovery of Rotruck et al. (1973) that selenium is a component of rat erythrocyte glutathione peroxidase. Further studies in this laboratory have demonstrated the important role of glutathione peroxidase in protection against the vitamin E- and selenium-deficiency disease of chicks, exudative diathesis, which results from increased capillary permeability. Also shown were the activities of both dietary selenium and vitamin E in prevention of ascorbate-induced peroxidation in mitochondrial and microsomal preparations from chick liver. Recent results demonstrate that both selenium and vitamin E are required to protect hepatic mitochondria and microsomes from peroxidative degradation. Dietary requirements of the chick for both nutrients for this function have been determined: approximately 0.06 ppm selenium in the presence of adequate vitamin E; 30-50 IU vitamin E per kg in the presence of adequate selenium.


Research ethics dilemmas in thyroid disease.
Since research ethics dilemmas frequently fall outside the purview of the Institutional Review Board (IRB), we present three unique recent research ethics cases in thyroidology that demonstrate research ethics dilemmas. The cases presented raise questions surrounding epistemic/scientific integrity, publication ethics, and professional, and personal integrity. Research ethics dilemmas that fall outside the purview of the IRB are appropriate for a Research Ethics Consultation, a common service in many large academic medical centers.


Circumvention of acquired tetraplatin resistance in a human ovarian carcinoma cell line by a novel trans platinum complex, JM335 [trans ammine (cyclohexylamine) dichloro dihydroxo platinum (IV)].
Acquired resistance to tetraplatin [d,1-trans-1,2-diaminocy-clohexane tetrachloroplatinum (IV)] has been generated in vitro in the human ovarian carcinoma cell line PXN94; the derived line, PXN94tetR, was 24-fold resistant to tetraplatin. Intracellular tetraplatin accumulation was reduced in PXN94tetR compared with PXN94 by an average of 1.3-fold across the concentration range 1-100 microM (2 hr exposure). There was no significant difference in glutathione levels between the 2 cell lines. PXN94tetR was 1.6-fold more resistant to cadmium chloride than PXN94, suggesting that metallothionein levels may be elevated. However, no significant difference was observed between PXN94 and PXN94tetR in the levels of total platinum bound to DNA or DNA interstrand cross-links immediately after tetraplatin exposure (10-100 microM x 2 hr). There was also no significant difference between the 2 cell lines in the rate of removal of total platinum or interstrand cross-links from DNA following 2 hr exposure to 25 microM tetraplatin. Hence the major mechanism of acquired tetraplatin resistance in PXN94tetR appears to be increased tolerance of platinum-DNA adducts. PXN94tetR was partially cross-resistant to the bifunctional alkylating agents melphalan, chlorambucil and mitomycin C. Partial cross-resistance was also observed to Adriamycin, bleomycin, etoposide, 5-fluorouracil and vinblastine; however, no elevation in P-glycoprotein levels was apparent in PXN94tetR. No cross-resistance was observed to taxotere. PXN94tetR was partially cross-resistant to cisplatin, carboplatin and several novel cis platinum complexes. In contrast, resistance was completely circumvented by the novel trans platinum complex JM335 [trans ammine (cyclohexylamine) dichloro dihydroxo platinum (IV)].


A Convenient Synthesis of Hydroxytyrosol Monosulfate Metabolites.
The growing interest in the bioactivity of natural polyphenols and of their metabolites requires metabolites to be used in bioassays and as standards in research protocols. We report here on the synthesis of several hydroxytyrosol metabolite monosulfates achieved using a simplified protocol with improved yields. A synthetic solution based on avoidance of high temperature conditions during the synthesis and of low pressure conditions during purification has been established. Monosulfates of several phenolic compounds, namely, hydroxytyrosol, hydroxytyrosol acetate, homovanillyl alcohol, homovanillyl alcohol acetate, homovanillic acid, ferulic acid, and 3,4-dihydroxyphenylethanoic acid, were efficiently synthesized in 1-2 steps in good yield and isolated using simple procedures. The proposed protocol was shown to be relatively rapid, efficient, cheap, and widely applicable to a number of catechol scaffolds.


Dehydroepiandrosterone sulfate reduces acoustic injury of the guinea-pig cochlea.
The present study was performed to determine effects of dehydroepiandrosterone sulfate (DHEAS), a neurosteroid, on acoustic injury. Albino guinea pigs were exposed to a 2 kHz pure tone of 120 or 125 dB sound pressure level for 10 min immediately after intravenous administration of DHEAS. Statistically significant improvement in the compound action potential threshold shifts and in amplitude reduction of distortion-product otoacoustic emissions was observed 1 week after the acoustic overexposure in the animals treated with DHEAS. The present results suggest that DHEAS has a protective effect against acoustic injury of the cochlea.


Attainment of Functional and Social Independence in Adult Survivors of Pediatric CNS Tumors: A Report From the St Jude Lifetime Cohort Study.
Purpose Beyond survival, achieving independence is a primary goal for adult survivors of pediatric CNS tumors. However, the prevalence of and risk factors for failure to achieve independence, assessed with multiple concurrent indicators, have not been examined. Patients and Methods Functional and social independence was assessed in 306 survivors (astrocytoma [n = 130], medulloblastoma [n = 77], ependymoma [n = 36], and other [n = 63]; median current age, 25.3 years [range, 18.9 to 53.1 years]; time since diagnosis, 16.8 years [range, 10.6 to 41.8 years]). Six observed indicators were used to identify latent classes of independence, which included employment, living independently, assistance with personal care, assistance with routine needs, obtaining a driver's license, and marital status. Physical performance impairments were defined as scores < 10th percentile on measures of aerobic capacity, strength, flexibility, balance, mobility, and adaptive function. Multinomial logistic regression estimated odds ratios (ORs) and 95% CIs were calculated for associations of disease/treatment exposures and impairments in physical performance with nonindependence. Results Three classes of independence were identified as independent (40%), moderately independent (34%), and nonindependent (26%). In multivariable models, craniospinal irradiation (OR, 4.20; 95% CI, 1.69 to 10.44) and younger age at diagnosis (OR, 1.24; 95% CI, 1.14 to 1.35) were associated with risk of nonindependence versus independence. Beyond impaired IQ, limitations in aerobic capacity (OR, 5.47; 95% CI, 1.78 to 16.76), flexibility (OR, 3.66; 95% CI, 1.11 to 12.03), and adaptive physical function (OR, 11.54; 95% CI, 3.57 to 37.27) were associated with nonindependence versus independence. Nonindependent survivors reported reduced physical but not mental health-related quality of life compared with independent survivors. Conclusion Sixty percent of survivors of pediatric CNS tumors do not achieve complete independence as adults. Reduction in intensity of primary therapies and interventions that target physical performance and adaptive deficits may help survivors to achieve greater independence.


[18F]choline positron emission tomography in prostate cancer patients with biochemical recurrence after radical prostatectomy: influence of antiandrogen therapy - a preliminary study.
Our purpose was to evaluate whether antiandrogen therapy (AAT) influences [(18)F]choline PET results in patients with biochemical recurrence after radical prostatectomy (RPE). Through a retrospective study we evaluated two groups of patients, both with histologically proven carcinoma of the prostate, who had undergone RPE and a subsequent [(18)F]choline PET because of biochemical failure (<4 ng/dl). One group consisted of 13 patients under AAT at the time of the PET examination (age range, 55-80 years; median, 68). The other group who had not undergone AAT consisted of 22 patients (age range, 48-72 years; median, 67). Our results were correlated with follow-up information related to histopathology, changes in prostate-specific antigen levels, other imaging modalities and clinical examination. Mean follow-up was 27 months. In patients who had undergone AAT, [(18)F]choline PET was true positive in eight out of 10 patients. The overall sensitivity in this group was 80%. In two cases [(18)F]choline PET turned out to be false negative, missing local relapse. Of the patients treated only with RPE, 10 out of 20 turned out to be true positive, resulting in a sensitivity of 50%.In all, in four patients biochemical recurrence could not be correlated to pathological findings in any of the available modalities. The difference in sensitivity between patients with and without AAT was statistically not significant (P=0.235). In patients with biochemical recurrence during AAT after RPE, [(18)F]choline PET can yield true-positive findings, even at prostate-specific antigen values of less than 4 ng/dl, and is an accurate technique for the detection of recurrence.


Topiramate moderately reduces the motivation to consume alcohol and has a marked antidepressant effect in rats.
In recent human studies, the anticonvulsant drug topiramate (TPM) has shown efficacy in treating alcohol craving and mood disorders. However, preclinical evidence supporting such effects is surprisingly sparse. Three experiments were conducted here to assess possible anticraving and antidepressant effects of TPM using animal models. In Experiment 1, rats were given 23 weeks ad libitum access to food, water, and either beer (4.44% ethanol v/v) or "near-beer" (a calorie-matched nonalcoholic beer, 0.44% ethanol) in their home cages. They were then restricted to daily 1 hour operant sessions in which they licked for water and either beer or near-beer under a progressive ratio schedule of reinforcement in a lickometer apparatus. The acute effects of TPM on the motivation to consume beer or near-beer were then assessed. The effects of naloxone were also assessed (as a positive control) after TPM testing. In Experiment 2, rats were given 11 weeks of ad libitum home-cage access to food, water, and beer. They then received repeated daily injections of TPM and effects on beer consumption under ad libitum home cage access conditions were monitored. In Experiment 3, the effects of TPM were assessed in the modified Porsolt forced swim test, emergence test, and elevated plus-maze (EPM) using alcohol naïve rats. Topiramate (10, 20, and 40 mg/kg) significantly reduced the motivation to lick for beer, although the maximal effect was moderate in comparison with naloxone (10 mg/kg). However, naloxone, unlike TPM, also reduced responding for near-beer suggesting an alcohol-specific effect of TPM. In Experiment 2, TPM (40 and 80 mg/kg) tended to transiently reduce alcohol consumption in the home cage under ad libitum access but this effect disappeared with repeated administration of the drug. TPM (10 to 80 mg/kg, given twice over 4 hours before test) produced a robust dose-dependent decrease in immobility and increase in active coping strategies in the forced swim test similar to that seen with desipramine (2 x 20 mg/kg). There were modest anxiolytic effects of TPM on the EPM and emergence tests. With acute administration, TPM is moderately effective and relatively selective in reducing the drive to consume alcohol in Wistar rats. This anti-alcohol effect is modest in comparison with naloxone and appears to dissipate under conditions of chronic treatment and ad libitum alcohol access. A marked antidepressant-like effect in the forced swim test and partial anxiolytic effects in other animal models suggests that TPM may be a beneficial treatment for affective disorders. These preliminary results suggest further research is warranted to resolve the mechanisms involved in TPM modulation of both mood and alcohol consumption.


Antistreptolysin O, rheumatoid factor and C-reactive protein determination in patients with recurrent oral ulcer.
Recurrent oral ulcer (ROU) is a common disease whose the etiologic factors have not yet been identified. Although the autoimmune mechanism due to streptococcal antigens may be involved in this disease, serum immunoglobulins are reported to be higher than normal controls in various phases of ROU. Oral ulcers are one of the criteria in the diagnosis of some systemic diseases such as Wegener's granulomatosis, Behçet's syndrome, suppurative arthritis, Reiter's syndrome and neutropenias. Any patient with recurring oral ulcers should be evaluated medically for the possible presence or future development of serious systemic diseases. In this study of the 58% of the ROU patients who had positive ASO values, only one female patient had a moderate increase: ++ASO(= 400 IU), the rest had a slight increase: +ASO(= 200 IU). In the control group only 3 female subjects had +ASO (= 200 IU). The aim of this study was to determine whether there is any relationship between ROU and rheumatoid arthritis. Patients with ROU were examined for antistreptolysin O, rheumatoid factor and C-reactive protein and compared to healthy controls.


Evidence for a spontaneous gapped state in ultraclean bilayer graphene.
At the charge neutrality point, bilayer graphene (BLG) is strongly susceptible to electronic interactions and is expected to undergo a phase transition to a state with spontaneously broken symmetries. By systematically investigating a large number of single-and double-gated BLG devices, we observe a bimodal distribution of minimum conductivities at the charge neutrality point. Although σ(min) is often approximately 2-3 e(2)/h (where e is the electron charge and h is Planck's constant), it is several orders of magnitude smaller in BLG devices that have both high mobility and low extrinsic doping. The insulating state in the latter samples appears below a transition temperature T(c) of approximately 5 K and has a T = 0 energy gap of approximately 3 meV. Transitions between these different states can be tuned by adjusting disorder or carrier density.


Analysis of Fourteen New Cases of Meningovascular Syphilis: Renewed Interest in an Old Problem.
Neurosyphilis (NS) remains a public health problem. Several recent reports suggest a worldwide increase in the incidence of this condition. Various syndromes can occur in NS, such as syphilitic meningitis, meningovascular syphilis, parenchymatous and gummatous neurosyphilis. Syphilis meningovascular will be the focus of this study. We report 14 new observations of meningovascular syphilis. A review of demographic and clinical features, neuroimaging findings, cerebrospinal fluid changes, treatment and outcome, pathophysiology mechanism of meningovascular syphilis are presented.


Detection of liver metastases with superparamagnetic iron oxide in 15 patients: results of MR imaging at 1.5 T.
The first clinical results of using superparamagnetic ferrite particles as a tissue-specific contrast agent for MR of the liver are reported at high fields (1.5 T). Fifteen patients with proved secondary liver malignancies were studied with plain and contrast-enhanced MR. Superparamagnetic iron oxide was administrated IV in a dose of 20 mumol/kg. Intermediate TR, 820/30, 60 (TR/TE), and long TR, 2200/22, 70, spin-echo sequences were used before and 1 hr after injection of contrast material. Before injection, the largest number of lesions (437) was detected with the T2-weighted sequence. Lesion-to-liver contrast, expressed as the difference between the tumor and liver signal-to-noise, improved after ferrite administration in both sequences from -1 to 20 and from 7 to 15 for the 820/30, 60 sequence and from 9 to 34 and 15 to 21 for the 2200/22, 70 sequence. Despite this significant improvement in terms of the number of lesions detected, contrast-enhanced images did not show significantly more metastases than the unenhanced T2-weighted images did (383, 421, 407, and 407 vs as many as 437, respectively). In this limited study at 1.5 T, the benefit of ferrite enhancement was only marginal when postcontrast images were compared with heavily T2-weighted precontrast scans.


Expression and regulation of Spätzle-processing enzyme in Drosophila.
The Drosophila melanogaster Toll receptor controls embryonic dorsal-ventral axis formation and is crucial for the innate immune response. In both cases, Toll is activated by the enzymatically cleaved form of its ligand Spätzle (Spz). During axis formation, Spz is cleaved by the maternally provided serine protease Easter while the Spätzle-processing enzyme (SPE) activates Spz after infection. We confirm the role of SPE in immunity and show that it is a zygotic gene specifically expressed in immune tissues implying that the dual activation of Spz is achieved by differential spatiotemporal expression of two similar but distinct serine proteases.


Primary arteritis of the aorta and its branches.
Primary arteritis of the aorta and its branches is a single clinicopathological entity affecting one or more segments of the aorta and resulting in a variety of symptom complexes. The prevalence is not known, but it is an important cause of renovascular hypertension in the black population of South Africa. Recently published articles on primary arteritis of the aorta and its branches are reviewed, and an attempt is made to draw conclusions regarding its cause, clinical features and treatment.


Genetic toxicology: current status of methods of carcinogen identification.
A critical aspect of the efforts to relate the results of short-term genetic toxicity tests with those from long-term rodent tests for carcinogens is the quality and consistency of the studies conducted by the National Toxicology Program. Analysis of the results in relationship to chemical structure has shown that mutagenic potential is a primary risk factor for carcinogen identification. Chemicals positive in the Salmonella assay-generally possess "structural alerts" for electrophilic interactions, are predominantly represented among chemicals producing trans-species carcinogenic effects in rodents, and among those identified as carcinogenic to humans. Current efforts are aimed at defining toxicological, structural, and mechanistic properties of nonmutagens that are carcinogenic in rodents.


Individual differences in non-clinical maternal depression impact infant affect and behavior during the still-face paradigm across the first year.
Maternal depression can significantly impact mothers' sensitivity to their infants' needs as well as infants' social and emotional development. The still-face paradigm (SFP) is widely used to assess infants' understanding of the contingency between their own behavior and that of their caregivers, as well as infants' ability to self-regulate arousal levels during sudden changes in maternal responsiveness. Infants of clinically depressed mothers display blunted levels of negative affect compared to infants of non-depressed mothers during the still-face (SF) phase. However, little is known about whether individual differences in elevated, non-clinical levels of maternal depression similarly affect mother-infant interactions. The current study examines the longitudinal effects of non-clinical maternal depression on infant and maternal behaviors during the SFP. Infants (N=63) were assessed at 5 and 9 months and maternal depression was assessed at 5 months using the Beck Depression Inventory (BDI). Infants of mothers with elevated levels of depression displayed less negative engagement during the SF phase compared to infants of mothers with lower levels of depression. This effect was present at 5 months, but not at 9 months. Findings demonstrate that non-clinical levels of maternal depressive symptomatology can have a significant impact on infants' affective regulation during the first half of the first year of life, but this does not necessarily have a long-lasting influence later in infancy. Interventions may want to target mothers with non-clinical depression to promote healthy infant social and emotional development.


Changes in gait parameters after femoral derotational osteotomy in cerebral palsy patients with medial femoral torsion.
Medial femoral torsion (MFT) is a common pathologic gait in cerebral palsy (CP) children that can be corrected by femoral derotational osteotomy (FDO). It is not clearly known as to how much various gait parameters change after FDO. The aim of this study was to quantify changes in gait parameters after FDO. The study group included 19 young CP patients (28 limbs, age<20 years, average age: 13.2 years) with symptomatic MFT, treated with distal FDO. The study group was divided into two groups: the unilateral FDO group (UG) and the bilateral FDO group (BG). The mean degree of derotation was 24.6° (25.0° for UG, 24.4° for BG). Pre-FDO and post-FDO values of Staheli's rotational profiles and kinematic data were compared. A paired t-test and Pearson's correlation were used for statistical analysis. The mean internal hip rotation was 71.4±6.9° before surgery and 48.6±10.7° after surgery in the UG (P<0.05) and it was 63.8±15.8° before surgery and 40.9±9.2° after FDO in the BG (P<0.05). The change in the foot progression angle (FPA) was 12.9° in the UG group (P<0.05) and 12.6° in the BG group (P<0.05). The degree of FPA had changed by about a half of the surgical derotation angle. Changes in the mean hip rotation during gait were 14.8° in the UG (P<0.05) and 6.7° in the BG (P<0.05) groups. The overall pelvic rotation was not changed after surgery. However, in patients with preoperative compensatory pelvic rotation of more than 5°, there was a change of 5.3±4.8° in the UG and 6.6±1.54° in the BG after surgery (P<0.05). There was also a trend showing that the younger the patient, the more the pelvic rotation changed (P=0.069). In-toeing gait because of MFT improved with FDO in CP patients. The expected degree of postoperative correction of FPA and hip rotation is about a half of the FDO degree. The degree of compensatory pelvic rotation should be considered to determine the correction angle of FDO, especially in young patients with preoperative pelvic rotation of more than 5°.


Intraocular lens power calculations in a patient with posterior keratoconus.
We describe a patient with posterior keratoconus accompanied by hyperopia and a cataract, which were successfully treated by cataract surgery and secondary piggyback intraocular lens (IOL) implantation. Case report. A 47-year-old woman presented to our clinic with low vision in her right eye. Uncorrected distant visual acuity was 20/400 in the right eye with a manifest refraction of +11.5 diopters. On the slit-lamp examination, localized posterior keratoconus and lens opacity were noted in the right eye. First, we performed phacoemulsification and posterior chamber IOL implantation. Postoperatively, hyperopic shift was noted because localized posterior elevation of the cornea is not considered in the conventional IOL power calculation. After secondary piggyback IOL implantation in the sulcus, her refractive error was finally properly corrected and uncorrected distant visual acuity improved to 20/50. In patients with posterior keratoconus, IOL power calculation from conventional keratometry may be inaccurate, and secondary piggyback IOL procedure may be needed after cataract surgery.


Hydration of guanidinium depends on its local environment.
Hydration of gaseous guanidinium (Gdm+) with up to 100 water molecules attached was investigated using infrared photodissociation spectroscopy in the hydrogen stretch region between 2900 and 3800 cm-1. Comparisons to IR spectra of low-energy computed structures indicate that at small cluster size, water interacts strongly with Gdm+ with three inner shell water molecules each accepting two hydrogen bonds from adjacent NH2 groups in Gdm+. Comparisons to results for tetramethylammonium (TMA+) and Na+ enable structural information for larger clusters to be obtained. The similarity in the bonded OH region for Gdm(H2O)20+vs. Gdm(H2O)100+ and the similarity in the bonded OH regions between Gdm+ and TMA+ but not Na+ for clusters with <50 water molecules indicate that Gdm+ does not significantly affect the hydrogen-bonding network of water molecules at large size. These results indicate that the hydration around Gdm+ changes for clusters with more than about eight water molecules to one in which inner shell water molecules only accept a single H-bond from Gdm+. More effective H-bonding drives this change in inner-shell water molecule binding to other water molecules. These results show that hydration of Gdm+ depends on its local environment, and that Gdm+ will interact with water even more strongly in an environment where water is partially excluded, such as the surface of a protein. This enhanced hydration in a limited solvation environment may provide new insights into the effectiveness of Gdm+ as a protein denaturant.


Current and Past Immunodeficiency Are Associated With Higher Hospitalization Rates Among Persons on Virologically Suppressive Antiretroviral Therapy for up to 11 Years.
Persons with human immunodeficiency virus (PWH) with persistently low CD4 counts despite efficacious antiretroviral therapy could have higher hospitalization risk. In 6 US and Canadian clinical cohorts, PWH with virologic suppression for ≥1 year in 2005-2015 were followed until virologic failure, loss to follow-up, death, or study end. Stratified by early (years 2-5) and long-term (years 6-11) suppression and lowest presuppression CD4 count <200 and ≥200 cells/µL, Poisson regression models estimated hospitalization incidence rate ratios (aIRRs) comparing patients by time-updated CD4 count category, adjusted for cohort, age, gender, calendar year, suppression duration, and lowest presuppression CD4 count. The 6997 included patients (19 980 person-years) were 81% cisgender men and 40% white. Among patients with lowest presuppression CD4 count <200 cells/μL (44%), patients with current CD4 count 200-350 vs >500 cells/μL had aIRRs of 1.44 during early suppression (95% confidence interval [CI], 1.01-2.06), and 1.67 (95% CI, 1.03-2.72) during long-term suppression. Among patients with lowest presuppression CD4 count ≥200 (56%), patients with current CD4 351-500 vs >500 cells/μL had an aIRR of 1.22 (95% CI, .93-1.60) during early suppression and 2.09 (95% CI, 1.18-3.70) during long-term suppression. Virologically suppressed patients with lower CD4 counts experienced higher hospitalization rates and could potentially benefit from targeted clinical management strategies.


Chaperone-mediated autophagy: Molecular mechanisms, biological functions, and diseases.
Chaperone-mediated autophagy (CMA) is a lysosomal degradation pathway that eliminates substrate proteins through heat-shock cognate protein 70 recognition and lysosome-associated membrane protein type 2A-assisted translocation. It is distinct from macroautophagy and microautophagy. In recent years, the regulatory mechanisms of CMA have been gradually enriched, including the newly discovered NRF2 and p38-TFEB signaling, as positive and negative regulatory pathways of CMA, respectively. Normal CMA activity is involved in the regulation of metabolism, aging, immunity, cell cycle, and other physiological processes, while CMA dysfunction may be involved in the occurrence of neurodegenerative disorders, tumors, intestinal disorders, atherosclerosis, and so on, which provides potential targets for the treatment and prediction of related diseases. This article describes the general process of CMA and its role in physiological activities and summarizes the connection between CMA and macroautophagy. In addition, human diseases that concern the dysfunction or protective role of CMA are discussed. Our review deepens the understanding of the mechanisms and physiological functions of CMA and provides a summary of past CMA research and a vision of future directions.


XP Shaper, A Novel Adaptive Core Rotary Instrument: Micro-computed Tomographic Analysis of Its Shaping Abilities.
The aim of this study was to investigate the shaping abilities of the XP Shaper (FKG, La Chaux-de-Fonds, Switzerland) and compare the findings with Vortex Blue (Dentsply Tulsa Dental Specialties, Tulsa, OK) using micro-computed tomographic imaging. Twenty matched, extracted, mandibular, central incisors with a single, oval canal were scanned preoperatively at 25-μm resolution and postoperatively after instrumentation with either Vortex Blue in a crown-down manner up to size 30.04 or XP Shaper. The percent of untouched walls, changes in canal volume and surface area, the amount of dentin removed, debris remaining in the canal, and the preparation taper were determined. The total time required for instrumentation using each technique was calculated in seconds. Statistical analysis was used to compare between both groups using repeated measures multivariate analysis of variance with Bonferroni correction for post hoc comparison and independent sample t tests. The XP Shaper significantly increased the canal volume (F = 77.948, P < .001), surface area (F = 5.543, P = .030), and amount of dentin removed (F = 10.044, P = .001) and had significantly less untouched walls (38.6% ± 8.1%) compared with VB (58.8% ± 8.5%). There was less debris at all levels of the canal in the XP Shaper group. Results were almost significant (P = .059). The XP Shaper was also significantly faster in completing the mechanical preparation of the root canal space by almost 1 minute (t = 6.216, P < .001). The XP Shaper can expand beyond its core size to adapt to the anatomy of the root canal space. The XP Shaper can prepare and touch more canal walls in oval-shaped canals compared with Vortex Blue. However, the final preparation taper will vary according to the anatomy of the treated tooth.


Pancreaticogastrostomy in patients with considerably dilated pancreatic ducts.
Patency of pancreaticogastrostomy (PG) is one of the most important factors affecting the function of the remnant pancreas and quality of life. We evaluated the early postoperative changes in pancreatic duct dilation after pancreaticoduodenectomy (PD) and PG with duct-to-mucosa anastomosis in patients with remarkably dilated pancreatic ducts. We retrospectively analyzed 26 patients who had remarkably dilated pancreatic ducts (diameter, ≥7 mm) and who underwent PD followed by PG. They were divided into 2 groups on the basis of the endoscopic findings of the anastomotic orifice of PG: Group A, clear pancreatic duct orifice with pancreatic juice output; and Group B, unclear pancreatic duct orifice with pancreatic juice output. The mean diameter of the duct of the remnant pancreas after the surgery was smaller in Group A than in Group B. With regards to postoperative pancreatic exocrine function, there was no significant difference between the 2 groups. Invagination with duct-to-mucosa anastomosis is a useful technique to prevent pancreatic leakage; however, it is difficult to prevent inflammation and fibrosis around the anastomotic site of PG, and this can lead to anastomotic stricture in patients with a remarkably dilated pancreatic duct (diameter ≥7 mm).


Platelet rich plasma and knee surgery.
In orthopaedic surgery and sports medicine, the knee joint has traditionally been considered the workhorse. The reconstruction of every damaged element in this joint is crucial in achieving the surgeon's goal to restore the knee function and prevent degeneration towards osteoarthritis. In the last fifteen years, the field of regenerative medicine is witnessing a boost of autologous blood-derived platelet rich plasma products (PRPs) application to effectively mimic and accelerate the tissue healing process. The scientific rationale behind PRPs is the delivery of growth factors, cytokines, and adhesive proteins present in platelets and plasma, as well as other biologically active proteins conveyed by the plasma such as fibrinogen, prothrombin, and fibronectin; with this biological engineering approach, new perspectives in knee surgery were opened. This work describes the use of PRP to construct and repair every single anatomical structure involved in knee surgery, detailing the process conducted in ligament, meniscal, and chondral surgery.


Conservation and alteration of HLA-B27-specific T cell epitopes on mouse cells. Implications for peptide-mediated alloreactivity.
Alloreactive CTL responses generate a great variety of clonal specificities. Such diversity may be related to recognition of multiple peptides constitutively bound to any given MHC alloantigen. Among human alloreactive CTL, only a fraction of the clones lyse mouse P815 cells expressing class I HLA proteins. In this study the fine specificity of HLA-B27 allorecognition on human or mouse cells by five human HLA-B27-specific CTL clones was comparatively analyzed. This was done to examine what degree of variation in epitope structure is compatible with recognition of HLA Ag on mouse cells. Nine site-specific HLA-B27 mutants were expressed on both human and mouse cells, after DNA-mediated gene transfer, to construct two analogous series of target cells. The reaction patterns of four of the five CTL clones with these cell panels were compatible with conservation of their corresponding epitopes upon expression of HLA-B27 on mouse cells. The reaction pattern of the fifth clone was different with either cell panel, indicating that its epitope was structurally altered on mouse cells. It also suggested a selectively increased expression of the determinant on these cells. The results suggest that most of the epitopes recognized by allospecific CTL clones reacting across species are either independent of any bound peptide or involve identical peptides from both cell types. However, some of these clones recognize alloantigen-bound peptides that are somewhat different in structure depending on the cell type, and may be expressed at the mouse cell surface in greater amounts. Such peptides could arise from related proteins in both species, and be polymorphic as a result of phylogenetic divergence.


Clearance mechanism of a mannosylated antibody-enzyme fusion protein used in experimental cancer therapy.
MFECP1 is a mannosylated antibody-enzyme fusion protein used in antibody-directed enzyme prodrug therapy (ADEPT). The antibody selectively targets tumor cells and the targeted enzyme converts a prodrug into a toxic drug. MFECP1 is obtained from expression in the yeast Pichia pastoris and produced to clinical grade. The P. pastoris-derived mannosylation of the fusion protein aids rapid normal tissue clearance required for successful ADEPT. The work presented provides evidence that MFECP1 is cleared by the endocytic and phagocytic mannose receptor (MR), which is known to bind to mannose-terminating glycans. MR-transfected fibroblast cells internalize MFECP1 as revealed by flow cytometry and confocal microscopy. Immunofluorescence microscopy shows that in vivo clearance in mice occurs predominantly by MR on liver sinusoidal endothelial cells, although MR is also expressed on adjacent Kupffer cells. In the spleen, MFECP1 is taken up by MR-expressing macrophages residing in the red pulp and not by dendritic cells which are found in the marginal zone and white pulp. Clearance can be inhibited in vivo by the MR inhibitor mannan as shown by increased enzyme activities in blood. The work improves understanding of interactions of MFECP1 with normal tissue, shows that glycosylation can be exploited in the design of recombinant anticancer therapeutics and opens the ways for optimizing pharmacokinetics of mannosylated recombinant therapeutics.


Early tracheotomy versus prolonged endotracheal intubation in unselected severely ill ICU patients.
Although several advantages are attributed to tracheotomy in ICU patients requiring mechanical ventilation (MV), true benefits and the optimal timing of tracheotomy remain controversial. In this study, we compared early tracheotomy (ET) with prolonged intubation (PI) in severely ill patients requiring prolonged MV. Prospective, randomized study. Twenty-five medical and surgical ICUs in France. Patients expected to require MV > 7 days. Patients were randomised to either (open or percutaneous) ET within 4 days or PI. The primary end-point was 28-day mortality. Secondary end-points were: the incidence of ICU-acquired pneumonia, number of d1-d28 ventilator-free days, time spent in the ICU, 60-day mortality, number of septic episodes, amount of sedation, comfort and laryngeal and tracheal complications. A sample size of 470 patients was considered necessary to obtain a reduction from 45 to 32% in 28-day mortality. After 30 months, 123 patients had been included (ET = 61, PI = 62) in 25 centres and the study was prematurely closed. All group characteristics were similar upon admission to ICU. No difference was found between the two groups for any of the primary or secondary end-points. Greater comfort was the sole benefit afforded by tracheotomy after subjective self-assessment by patients. The trial did not demonstrate any major benefit of tracheotomy in a general population of ICU patients, as suggested in a previous meta-analysis, but was underpowered to draw any firm conclusions. The potential advantage of ET may be restricted to selected groups of patients.


Effects of amine modification of mesoporous magnesium carbonate on controlled drug release.
(3-Aminopropyl)triethoxysilane (APTES) was used to modify the surface of mesoporous magnesium carbonate (MMC). The as-synthesized MMC had an average pore diameter of ∼5nm, but amine grafting occurred preferentially on the walls of the largest MMC pores. Analysis of ibuprofen (IBU) loading and release showed that IBU remained stable in the amorphous phase in all the MMC and modified MMC samples. The kinetics of IBU release from the modified MMC were assessed and used to evaluate the effects of the different functional groups. The release rate showed that the release of IBU could be controlled by adjusting the amine surface coverage of MMC and also by changing the surface groups. It was concluded that the interaction between the grafted functional groups in the modified MMC and the OH in the carboxyl groups of IBU was the most important factor for prolonging the release of the drug. These results are expected to lead to investigation of other as yet unexplored applications for MMC, including using it as a plastic additive and for gas separation.


Mutant B-RAF-Mcl-1 survival signaling depends on the STAT3 transcription factor.
Approximately 50% of melanomas depend on mutant B-RAF for proliferation, metastasis and survival. The inhibition of oncogenic B-RAF with highly targeted compounds has produced remarkable albeit short-lived clinical responses in B-RAF mutant melanoma patients. Reactivation of signaling downstream of B-RAF is frequently associated with acquired resistance to B-RAF inhibitors, and the identification of B-RAF targets may provide new strategies for managing melanoma. Oncogenic B-RAF(V600E) is known to promote the stabilizing phosphorylation of the anti-apoptotic protein Mcl-1, implicated in melanoma survival and chemoresistance. We now show that B-RAF(V600E) signaling also induces the transcription of Mcl-1 in melanocytes and melanoma. We demonstrate that activation of STAT3 serine-727 and tyrosine-705 phosphorylations is promoted by B-RAF(V600E) activity and that the Mcl-1 promoter is dependent on a STAT consensus-site for B-RAF-mediated activation. Consequently, suppression of STAT3 activity disrupted B-RAF(V600E)-mediated induction of Mcl-1 and reduced melanoma cell survival. We propose that STAT3 has a central role in the survival and contributes to chemoresistance of B-RAF(V600E) melanoma.


Effects of the ecto-ATPase apyrase on microglial ramification and surveillance reflect cell depolarization, not ATP depletion.
Microglia, the brain's innate immune cells, have highly motile processes which constantly survey the brain to detect infection, remove dying cells, and prune synapses during brain development. ATP released by tissue damage is known to attract microglial processes, but it is controversial whether an ambient level of ATP is needed to promote constant microglial surveillance in the normal brain. Applying the ATPase apyrase, an enzyme which hydrolyzes ATP and ADP, reduces microglial process ramification and surveillance, suggesting that ambient ATP/ADP maintains microglial surveillance. However, attempting to raise the level of ATP/ADP by blocking the endogenous ecto-ATPase (termed NTPDase1/CD39), which also hydrolyzes ATP/ADP, does not affect the cells' ramification or surveillance, nor their membrane currents, which respond to even small rises of extracellular [ATP] or [ADP] with the activation of K+ channels. This indicates a lack of detectable ambient ATP/ADP and ecto-ATPase activity, contradicting the results with apyrase. We resolve this contradiction by demonstrating that contamination of commercially available apyrase by a high K+ concentration reduces ramification and surveillance by depolarizing microglia. Exposure to the same K+ concentration (without apyrase added) reduced ramification and surveillance as with apyrase. Dialysis of apyrase to remove K+ retained its ATP-hydrolyzing activity but abolished the microglial depolarization and decrease of ramification produced by the undialyzed enzyme. Thus, applying apyrase affects microglia by an action independent of ATP, and no ambient purinergic signaling is required to maintain microglial ramification and surveillance. These results also have implications for hundreds of prior studies that employed apyrase to hydrolyze ATP/ADP.


Early Versus Delayed Mortality among HIV Infected Patients Initiating Highly Active Antiretroviral Therapy in Tanzania.
Development of HAART in the mid-1990's and its continued scale up has revolutionized the treatment of HIV-infected patients and led to remarkable reductions in HIV associated morbidity and mortality. However, recent studies have suggested a higher risk for early mortality in adults receiving ART in low-income countries as compared to those in high-income countries. There is dearth of data from developing countries where the burden of disease is high. The objective is to describe the burden and correlation between early vs. delayed mortality associated with HIV/AIDS in resource poor settings using data from Tanzania in East Africa. We performed a cross-sectional evaluation of routinely collected program data for 991 HIV-positive deceased adult patients who were placed on ART treatment, and died between January 1, 2007 and December 31, 2012. Data used were abstracted from records of patients who were treated at six health facilities in the Lake-zone Region of Tanzania in the timeframe. Bivariate and multivariate regression models were used to identify independent predictors of mortality and to calculate odds ratios. From the population, early deaths (within 3 months of ART initiation) occurred in 359 of the 991 cases, which represented 36.2%; while delayed deaths (after 3 months of ART initiation) occurred in 632 of 991 (63.8%). The average time to death for those who died within 3 months was 1 month compared to 22 months among those who died at > 3 months since initiation of ARV. In multivariate analysis, patients who were on WHO stage IV, had fever and cough symptoms at 6 months prior to death and patients with 0-1, 2-3, and 4-6 clinic visits had a higher risk of death in the first 3 months. Mortality among patients started on ART seems to be high. Where possible, healthcare providers should do more to vigorously monitor patients before starting them on ART for better outcomes. Additionally, public health efforts to encourage early testing and entry into treatment must be scaled up in resource poor countries to gain some lead-time and to keep the virus under control, sustain immune function, and delay the onset of opportunistic infections.


Ignition of eyelash extensions during routine minor eyelid surgery.
Preoperative patient questionnaires enquiring about the presence of prosthetics are routine. Eyelash extensions are not part of standard preoperative questionnaires. This case illustrates how this impacts patient safety during minor eyelid surgery.


Pulmonary Apical Opacities on Thin-Section Computed Tomography: Relationship to Primary Spontaneous Pneumothorax in Young Male Patients and Corresponding Histopathologic Findings.
The purpose of this study was to test the hypothesis that apical opacities on computed tomography (CT) are related to occurrence of primary spontaneous pneumothorax (PSP) in young male patients. We compared the frequency of apical opacities on thin-section CT between 70 male patients with PSP (PSP group) and 74 male patients without a history of PSP (non-PSP group). We also evaluated histopathologic findings of 39 specimens from 37 surgical cases in the PSP group. Apical opacities were significantly more frequent in the PSP group than in the non-PSP group (right side, P = 0.01; left side, P = 0.005). Histopathologically, subpleural band-like alveolar collapse was seen in 35 specimens (89.7%), which was always accompanied by fibroelastosis and fibroblastic foci. Apical opacities on CT were significantly associated with PSP in young male patients. These apical opacities histopathologically correspond to fibrotic pleural thickening with subpleural alveolar collapse.


Self-reports of anxiety in burn-injured hospitalized adults during routine wound care.
The purpose of this investigation was to examine the amount of anxiety patients believed tolerable and the amount of anxiety experienced during routine burn wound care. Participants included 47 hospitalized adults who provided data for four consecutive assessment periods. Patients (mean TBSA, 16%; range, 2-70%) were primarily Caucasian (87%) and had an average hospital stays of 23 days (range, 11-130). Reports of what level of anxiety they would be able to tolerate and what level of anxiety had been experienced were assessed using 10-point Graphic Rating Scales. The use of anxiolytic was recorded, and patient suggestions for reducing anxiety were obtained. The single most commonly endorsed anxiety treatment goal was 0, although 53% consistently chose a treatment goal other than 0 (range, 1-6). Two repeated-measure analyses of variance indicated that the amount of anxiety patients could tolerate and the amount they reported experiencing did not change over the course of time. Paired t-tests revealed that patients routinely reported more anxiety than they considered tolerable. Analyses of anxiety reports of patients treated with anxiolytics (n = 6) vs patients receiving no anxiolytics (n = 41) revealed inconsistent differences in actual anxiety and treatment goals across time. In general, patient suggestions for lessening anxiety included requests for education, communication, additional medications, and manipulation of the hospital environment. Anxiety for burn-injured, hospitalized adults remains a concern. Our findings are consistent with the literature indicating that adult patients hospitalized for burn wound care report appreciable anxiety, over and above what they consider "tolerable." Continued research is needed and should include investigations into the relationship between pain and anxiety during routine wound care.


A de novo deletion at 16q24.3 involving ANKRD11 in a Japanese patient with KBG syndrome.
KBG syndrome is a rare autosomal dominant congenital syndrome comprising developmental delay with various neurological involvements, macrodontia of the upper central incisors, characteristic facial dysmorphism, and skeletal anomalies. ANKRD11 was recently identified as the gene responsible for this syndrome. To date, there have been only five KBG syndrome families described, each carrying a single base substitution or a 1- to 14-bp deletion of this gene. Here, we present a patient with clinically confirmed KBG syndrome carrying a de novo 690-kb deletion at 16q24.3 involving part of ANKRD11. He had characteristic facial appearance, macrodontia of the upper central incisors, hand anomalies, delayed bone age and intellectual impairment without autistic features. Interestingly, the deleted region overlaps with the critical region for 16q24.3 microdeletion syndrome. We discuss the clinical entities of KBG syndrome and 16q24.3 microdeletion syndrome from a clinical and genetic point of view.


Dysphagia after arteria lusoria dextra surgery: Anatomical considerations before redo-surgery.
Aberrant right subclavian artery (arteria lusoria) is the most common congenital root anomaly, remaining asymptomatic in most cases. Nevertheless, some of the 20%-40% of those affected present tracheo-esophageal symptoms. We report on a 6-year-old previously healthy girl presenting with progressive dysphagia over 4 wk. Diagnostics including barium swallow, echocardiography and magnetic resonance angiography (MRA) revealed a retro-esophageal compression by an aberrant right subclavian artery. Despite the successful, uneventful transposition of this arteria lusoria to the right common carotid via right-sided thoracotomy, the girl was suffering from persisting dysphagia. Another barium swallow showed the persistent compression of the esophagus on the level where the arteria lusoria had originated. As MRA showed no evidence of a significant re-obstruction by the transected vascular stump, we suspected a persisting ligamentum arteriosum. After a second surgical intervention via left-sided thoracotomy consisting of transecting the obviously persisting ligamentum and shortening the remaining arterial stump of the aberrant right subclavian artery, the patient recovered fully. In this case report we discuss the potential relevance of a persisting ligamentum arteriosum for patients with left aortic arch suffering from dysphagia lusoria and rational means of diagnosing, as well as the surgical options to prevent re-do surgery.


Scaleup of monoclonal antibody purification using radial streaming ion exchange chromatography.
A scaleup study of the radial streaming chromatography (ZetaPrep technique) using hybridoma culture supernatant as model protein solution is described in this article. Lab and pilot cartridges were tested. Scaleup factors were calculated from the lab experiments and compared to the data obtained at pilot level. The procedure consists of three different steps: microfiltration, diafiltration, and the ZetaPrep technique using QAE cartridges. Diafiltration was used to condition the clarified culture supernatant. Calculating the elution volumes for the pilot level (ZetaPrep 800) from the smallest lab cartridge (ZetaPrep 15), a difference between calculated and experimental values of 230% was obtained. The difference between calculated and experimental values using results from ZetaPrep 100, a preparative cartridge, was 120%. At pilot level it is possible to purify 10 L culture supernatant within 3 h including regeneration and reequilibration of the cartridge. This procedure is useful for monoclonal antibodies (mAb) with a low isoelectric point (pl). The pl's of the mAb which was used in this work are in the range 5.4-6.1.


Long-term cardiovascular outcomes and overall survival of early-stage breast cancer patients with early discontinuation of trastuzumab: a population-based study.
We critically examined long-term cardiovascular (CV) outcomes and overall survival (OS) of breast cancer (BC) patients who had cardiotoxicity during adjuvant trastuzumab treatment requiring discontinuation in a population-based sample. This was a retrospective cohort of early-stage BC patients diagnosed before 2010 and treated with trastuzumab in Ontario. Patients were stratified based on trastuzumab doses received: 1-8, 9-15, ≥16 (therapy completion). Time-dependent multivariable Cox models were used to analyze primary endpoint OS, and the following composite endpoints: hospitalization/emergency room visit for heart failure (HF) or death; non-HF CV (myocardial infarction, stroke) or death; and clinically significant relapse (palliative systemic therapy initiation >90 days after last trastuzumab dose) or death. Of the 3134 women, 6, 10, and 85 % received 1-8, 9-15, and ≥16 doses, respectively. Over 5-year median follow-up, early trastuzumab discontinuation was associated with more HF/death [1-8 doses hazard ratio (HR) 4.0, 95 % confidence interval (CI) 2.7-6.0; 9-15 doses HR 2.97, 95 % CI 2.1-4.3], non-HF/death (1-8 doses HR 4.3, 95 % CI 3.0-6.1; 9-15 doses HR 3.1, 95 % CI 2.2-4.4), clinically significant relapse/death (1-8 doses HR 3.1, 95 % CI 2.2-4.4; 9-15 doses HR 2.4, 95 % CI 1.8-3.3), and importantly lower OS (77, 80, 93 %; P < 0.001). Early discontinuation (1-8 doses HR 2.41, 95 % CI 1.5-3.8; 9-15 doses HR 2.9, 95 % CI 2.0-4.1) and clinically significant relapse (HR 34.0, 95 % CI 24.9-46.6) were both independent predictors of mortality. Of note, early discontinuation remained a critical independent predictor of OS even after adjusting for incident HF. Early trastuzumab discontinuation is a powerful independent predictor of cardiac events and clinically significant relapse, and both may contribute to poor survival. Both adequate cancer control and optimal CV management are required to improve long-term outcomes.


In silico screening and identification of potential drug against p300 acetyltransferase activity in breast cancer via drug repurposing approach.
Emerging evidence portray the involvement of epigenomic reprogramming in the onset and progression of several malignancies, including breast cancer. Histone acetyltransferase (HAT) p300 is a critical epigenetic regulator that acts as a transcription co-activator and regulates various cellular processes. p300 is overexpressed in breast cancer and promotes cellular invasion and survival, making it a promising druggable target. In this study, the relevance of p300 in different cancer pathways was established. Virtual screening of the FDA-approved drug library was carried out using molecular docking, and the top 10 potential repurposed drugs were identified. Further, recalculation of binding free energy of drug-p300 complexes was carried out using molecular mechanics Poisson-Boltzmann and surface area (MM-PBSA) method after molecular dynamic simulation. Based on molecular dynamic simulation parameters and binding free energy analysis, two drugs, namely Netarsudil (-305.068 kJ/mol) and Imatinib (-260.457 kJ/mol), were identified as potential repurposed drugs to inhibit the activity of p300. In conclusion, these findings suggest, Netarsudil and Imatinib might be a potential repurposed drug to combat breast cancer via p300 inhibition.Communicated by Ramaswamy H. Sarma.


Vancomycin-resistant enterococci in acute myeloid leukemia and myelodysplastic syndrome patients undergoing induction chemotherapy with idarubicin and cytarabine.
We conducted a retrospective study to determine the risk factors associated with vancomycin-resistant enterococci (VRE) acquisition/infection in newly diagnosed acute myeloid leukemia and myelodysplastic syndrome patients undergoing chemotherapy with the 7 + 3 regimen of cytarabine and idarubicin. Although only 2.5% (6/235) patients were colonized with VRE on admission, 59% (134/229) of patients acquired VRE during their hospitalization. Multivariable analysis identified the use of intravenous vancomycin (p = .024; HR: 1.548) and cephalosporin (p = .009; HR: 1.596) as the risk factors for VRE acquisition. VRE infection developed in 14% (33/229) of patients, with bloodstream infections accounting for 82% (27/33) of cases. VRE infection occurred in 25/126 (20%) of the VRE-colonized patients, but only 8/103 (8%) of those who were not (p = .01). Our study provides the evidence for the role of intravenous cephalosporin and vancomycin in VRE acquisition and highlights the clinical significance of VRE colonization in these patients.


Pd-catalyzed intramolecular aerobic oxidative C-H amination of 2-aryl-3-(arylamino)quinazolinones: synthesis of fluorescent indazolo[3,2-b]quinazolinones.
A palladium-catalyzed intramolecular aerobic oxidative C-H amination of 2-aryl-3-(arylamino)quinazolinones has been developed, providing a variety of substituted indazolo[3,2-b]quinazolinone derivatives in moderate to excellent yields. Preliminary mechanistic studies suggested that a palladacycle dimer could be the key intermediate, which underwent a cascade "rollover" cyclometalation and C-H amination sequence. Furthermore, the potential utility of these products has been demonstrated as a new class of blue fluorophores for fluorescent materials.


The origin recognition complex marks a replication origin in the human TOP1 gene promoter.
The locations of the origin recognition complex (ORC) in mammalian genomes have been elusive. We have therefore analyzed the DNA sequences associated with human ORC via in vivo cross-linking and chromatin immunoprecipitation. Antibodies specific for hOrc2 protein precipitate chromatin fragments that also contain other ORC proteins, suggesting that the proteins form multisubunit complexes on chromatin in vivo. A binding region for ORC was identified at the CpG island upstream of the human TOP1 gene. Nascent strand abundance assays show that the ORC binding region coincides with an origin of bidirectional replication. The TOP1 gene includes two well characterized matrix attachment regions. The matrix attachment region elements analyzed contain no ORC and constitute no sites for replication initiation. In initial attempts to use the chromatin immunoprecipitation technique for the identification of additional ORC sites in the human genome, we isolated a sequence close to another actively transcribed gene (TOM1) and an alphoid satellite sequence that underlies centromeric heterochromatin. Nascent strand abundance assays gave no indication that the heterochromatin sequence serves as a replication initiation site, suggesting that an ORC on this site may perform functions other than replication initiation.


Double-chambered right ventricle associated with infective endocarditis complication of the pulmonary valve: surgical management.
Double-chambered right ventricle (DCRV) is a rare congenital heart disease characterized by the presence of anomalous muscle bundles, which divide the right ventricle into two chambers: a high-pressure proximal chamber and a low-pressure distal chamber. Most DCRV patients are diagnosed and treated during childhood, and presentation in adulthood is not common. Many congenital heart diseases are often associated with other complications such as infective endocarditis (IE). Right-side endocarditis, which usually involves infection of the tricuspid valve, is uncommon, and endocarditis of the pulmonary valve is extremely rare. We report a 51-year-old woman with undiagnosed DCRV and ventricular septal defect associated with pulmonary valve endocarditis. The diagnostic evaluation and the surgical management are discussed.


The Nurses' Global Assessment of Suicide Risk (NGASR): developing a tool for clinical practice.
Contemporary and established literature indicates that people with mental health problems are at a higher risk of suicide than the general population. Because suicide is a multifaceted, complex phenomenon, risk assessment within the mental health care system requires a pluralistic, multidimensional and multiprofessional response. While assessment tools may provide useful guidance, especially guarding against complacency and over confidence, the fundamental basis of risk assessment must involve a thorough examination of the personal, interpersonal and social circumstances of each individual. Such thorough and rigorous assessments, the authors of this paper would add, require a degree of 'clinical judgement'. As a rule, inexperienced members of mental health care staff should not be charged with the responsibility of conducting suicide risk assessments without sound mentorship. However, with the right support and assessment tool, the novice practitioner might develop the kind of clinical judgement necessary for this critical task. Accordingly, this paper traces the development of the Nurses' Global Assessment of Suicide Risk (NGASR). It illustrates the practice development context out of which the need for the tool arose; it outlines the key evidence that underpins the construction of the tool and it is described. It is important to point out that as yet, no wide scale, quantitative validation of the tool has been conducted. Therefore, at this point, the tool should be treated with a degree of appropriate caution. Nevertheless, the preliminary attempts that have been made to 'validate' or 'rate' the tool in practice are included. While acknowledging that any risk assessment tool represents only one aspect of the necessarily broader assessment of risk, the NGASR appears to provide a useful template for the nursing assessment of suicide risk, especially for the novice.


Cerebral function monitor in the neonate. II: Birth asphyxia.
The cerebral function monitor (CFM) records an integrated electroencephalogram on slow-running paper, and therefore is suited to long-term, continuous monitoring. This study describes CFM patterns of 31 neonates with birth asphyxia. Three distinct types emerged: (1) a normal pattern compatible with gestational age was uniformly associated with favorable clinical outcome; (2) a completely disorganized pattern was associated with sever injury and fatal outcome; and (3) a more subtle pattern showed reversal to a more immature gestational age. The three infants with the third pattern all survived, but with varying degrees of neurological deficit. It is concluded that the CFM can be of advantage in predicting outcome for asphyxiated neonates.


Electronic and optical properties of the hydrogen boride sheet from the many-body perturbation theory.
We study the electronic and optical properties of the hydrogen boride sheet by using the many-body perturbation theory with the perturbativeGW(G0W0) approximation. It was found that the hydrogen boride sheet shows a semimetallic electronic structure, supporting the previous theoretical study based on the semilocal density functional theory calculations. It was also found that the optical spectrum calculated based on the quasiparticle energies agrees well with the experiments. This work suggests thatG0W0approximation may be useful for predicting precise electronic and optical properties of the hydrogen boride sheet and its derivatives.


Phospholipids and their degrading enzyme in the tears of soft contact lens wearers.
Low tear phospholipids levels are associated with tear film instability in soft contact lens wearers. We assayed levels of phospholipids and their degrading enzyme secretory phospholipase A2 (sPLA2) both in tears and deposited on contact lenses composed of 2 hydrophilic materials after 1 day of routine use. Polymacon (Medalist; FDA group 1, low water/nonionic) and Etafilcon A (One Day Acuvue; group 4, high water/ionic) contact lenses were worn for 12 hours by 16 experienced contact lens wearers. Phospholipids in tear fluids and deposited on contact lenses were estimated by phosphorus determination with ammonium molybdate through enzymatic digestion. Double-antibody sandwich ELISA was used to determine group IIa sPLA2 concentrations, and sPLA2 activity was assayed using 1,2-diheptanoyl thio-phosphatidylcholine as substrate. Phospholipids concentrations in tears with Polymacon and Etafilcon A were 186 +/- 39 and 162 +/- 33 microg/mL, respectively. The latter concentration was significantly lower than that observed in the same subjects when not wearing contact lenses (P = 0.0023). In tears, both group IIa sPLA2 concentrations and enzymatic activity remained unchanged, regardless of lens wearing. However, Etafilcon A (0.57 +/- 0.09 microg/lens) showed more group IIa sPLA2 deposition than Polymacon (0.01 +/- 0.01 microg/lens; P < 0.001). Furthermore, group IIa sPLA2 deposited on Etafilcon A but not on Polymacon lenses retained its enzymatic activity. Significant differences of group IIa sPLA2 deposition were found in the 2 lenses tested. Such deposition might induce phospholipid hydrolysis in tears and thereby promote tear film instability in hydrophilic contact lens wearers.


On randomly changing conformity bias in cultural transmission.
Humans and nonhuman animals display conformist as well as anticonformist biases in cultural transmission. Whereas many previous mathematical models have incorporated constant conformity coefficients, empirical research suggests that the extent of (anti)conformity in populations can change over time. We incorporate stochastic time-varying conformity coefficients into a widely used conformity model, which assumes a fixed number n of "role models" sampled by each individual. We also allow the number of role models to vary over time ([Formula: see text]). Under anticonformity, nonconvergence can occur in deterministic and stochastic models with different parameter values. Even if strong anticonformity may occur, if conformity or random copying (i.e., neither conformity nor anticonformity) is expected, there is convergence to one of the three equilibria seen in previous deterministic models of conformity. Moreover, this result is robust to stochastic variation in [Formula: see text] However, dynamic properties of these equilibria may be different from those in deterministic models. For example, with random conformity coefficients, all equilibria can be stochastically locally stable simultaneously. Finally, we study the effect of randomly changing weak selection. Allowing the level of conformity, the number of role models, and selection to vary stochastically may produce a more realistic representation of the wide range of group-level properties that can emerge under (anti)conformist biases. This promises to make interpretation of the effect of conformity on differences between populations, for example those connected by migration, rather difficult. Future research incorporating finite population sizes and migration would contribute added realism to these models.


Pattern and causes of amputation in diabetic patients--a multicentric study from India.
One of the most significant complications of diabetes is foot disease, which often leads to amputations was found to be very common in developing countries like India, the diabetic capital of the world. Hence this study has been planned to assess the pattern and causes of amputations in diabetic patients across various parts of India. A total of 1985 (M:F 1249:736) type 2 diabetic subjects were selected from 31 centres across India. Out of 1985 subjects, a total of 1295 (850:445) patients who had undergone amputations both major and minor were included in this analysis. A proforma which contains details on level of amputations, diabetes history, deformity details, causes of amputations and other associated diabetic complications was used to collect the data. Peripheral vascular disease was assessed by using Doppler studies. Presence of neuropathy was assessed by using 10 g monofilament and 125 Hz tuning fork. The major cause for the occurrence of amputations among the patients was found to be infection. Almost 90% of the patients had infection. Patients had different types of amputations: major amputations accounting for 29.1% (n=377) and minor amputations in 70.9% (n=918) of subjects. Among the subjects who underwent major amputations, more than 50% accounts for below knee amputations and 11.9% above knee amputations. Out of total amputations, over half of the incident amputations were of toes and rays. Presence of claw toes was seen in 64% of patients. Prevalence of neuropathy (82%) was high and 35% had peripheral vascular disease. In conclusion, infection was found to be the major cause of amputation in India. Below knee, toes and rays amputations were the most common type of amputations. Diabetic patients should be educated on foot care and importance of proper foot wear.


Phosphorylation of paxillin tyrosines 31 and 118 controls polarization and motility of lymphoid cells and is PMA-sensitive.
Tyrosine phosphorylation of paxillin regulates actin cytoskeleton-dependent changes in cell morphology and motility in adherent cells. In this report we investigated the involvement of paxillin tyrosine phosphorylation in the regulation of actin cytoskeleton-dependent polarization and motility of a non-adherent IL-3-dependent murine pre-B lymphocytic cell line Baf3. We also assessed the effect of phorbol myristate acetate (PMA), a phorbol ester analogous to those currently in clinical trials for the treatment of leukemia, on paxillin phosphorylation. Using tyrosine-to-phenylalanine phosphorylation mutants of paxillin and phosphospecific antibody we demonstrated that IL-3 stimulated phosphorylation of paxillin tyrosine residues 31 and 118, whereas the tyrosines 40 and 181 were constitutively phosphorylated. Phosphorylation of paxillin residues 31 and 118 was required for cell polarization and motility. In the presence of IL-3, PMA dramatically reduced the phosphorylation of residues 31 and 118, which was accompanied by inhibition of cell polarization and motility. This PMA effect was partially recapitulated by expression of exogenous tyrosine 31 and 118 mutants of paxillin. We also demonstrated that PMA inhibited the IL-3-induced and activation-dependent tyrosine phosphorylation of focal adhesion kinase. Thus, our results indicate that phosphorylation of paxillin tyrosine residues 31 and 118 regulates actin-dependent polarization and motility of pre-B Baf3 cells, both of which could be inhibited by PMA. They also suggest that inhibition of upstream signaling by PMA contributes to the decrease of paxillin phosphorylation and subsequent changes in cell morphology.


Sleep-Wake Behavior in Elite Athletes: A Mixed-Method Approach.
Using a mixed-method approach, this investigation aimed to examine athletes' sleeping patterns along with the socio-physiological acute and chronic stressors affecting their sleep. Fifteen elite athletes (M age = 21.7 years; SD = 2.2) were monitored daily during a typical in-season training period (20 ± 1 days) and completed individual semi-structured interviews. Their sleep was analyzed using actigraphy and sleep diaries. A content analysis method was used to assess qualitative interviews. Three factors influencing sleep emerged from the qualitative interview analyses, i.e., sleep environment, training and study requirements. Half the athletes (8/15) complained about their sleeping environment with noise and/or bedroom temperature and/or mattress quality and/or light exposure presented as an issue. "Complainers" notably exhibited impaired (p < 0.05) sleep efficiency and wake after sleep onset compared with "non-complainers." Daily training load showed correlations (p < 0.05) with wake after sleep onset and sleep onset latency. "Student athletes" exhibited later bedtime and earlier wake-up time compared with "non-student athletes," leading to a reduced total sleep time (6:50 ± 1:13 vs. 7:06 ± 0:53; p < 0.05; d = 0.20). An individualized assessment of sleep using actigraphy and interviews, with consideration to various socio-physiological factors, is recommended. Individualized sleep interventions with consideration to physiological (e.g., training load), behavioral (e.g., screen use) and environmental factors (e.g., room temperature, noise, mattress), can then be provided to each athlete.


N2-guanyl and N6-adenyl arylation of chicken erythrocyte DNA by the ultimate carcinogen of 4-nitroquinoline 1-oxide.
Great quantities of chicken erythrocyte DNA with high levels of modification were obtained in vitro by reaction with 4-acetoxyaminoquinoline 1-oxide, a model ultimate carcinogen of 4-nitroquinoline 1-oxide. After enzymatic hydrolysis of the modified DNA, the three main adducts were separated and isolated by semipreparative high performance liquid chromatography. These three adducts were already characterized in vivo and in vitro in our previous work (S. Galiègue-Zouitina et al., Cancer Res., 45: 520-525, 1985). The structure of one of them was previously identified as N-(deoxyguanosin-8-yl)-4-aminoquinoline 1-oxide (B. Bailleul et al., Cancer Res., 41: 4559-4565, 1981). In this paper we have identified by mass spectroscopy and nuclear magnetic resonance the structures of the two other main adducts as 3-(deoxyguanosin-N2-yl)-4-aminoquinoline 1-oxide and 3-(deoxyadenosin-N6-yl)-4-aminoquinoline 1-oxide, respectively.


Prenatal MRI evaluation of giant neck masses in preparation for the fetal exit procedure.
Airway compromise at birth can be anticipated with giant neck masses, so that as much anatomic information as possible is necessary prior to delivery to establish a diagnosis and assess the airway. With this study, we evaluated the usefulness of prenatal magnetic resonance imaging (MRI) for evaluation of giant fetal neck masses prior to operating on placental support, the Ex Utero Intrapartum Treatment (EXIT) procedure, performed to secure the neonatal airway. Three pregnant women with an ultrasound (US) diagnosis of fetal giant neck mass were referred for prenatal MRI. As this was our initial experience performing prenatal MRI, a variety of imaging sequences were used including spin-echo, fast gradient-echo, half-fourier single shot turbo spin-echo (Haste), and echo-planar imaging (EPI). All sequences performed were able to demonstrate the fetal airway relative to the mass. In addition, the images were able to give a more global definition of the mass because of the larger field of view than could be obtained with MRI compared to US. The Haste sequence provided the best definition of a mass because of decreased motion artifacts. The prenatal diagnosis on MRI was teratoma in two and lymphangioma in one and was confirmed at birth. MRI provided essential information about the diagnosis and the anatomy of the giant neck masses and adjacent airway in three fetuses prior to selection for the EXIT surgical procedure.


Release of (oligo) peptidyl-tRNA from ribosomes by erythromycin A.
Erythromycin A released peptidyl-tRNA in the in vitro polypeptide synthesis system with bacterial components programmed by synthetic polynucleotide. This is consistent with our hypothesis that erythromycin A inhibits translocation by preventing proper situation of oligopeptidyl-tRNA in the donor (D) site on ribosomes.


Neonatal Late Onset Infection with Severe Acute Respiratory Syndrome Coronavirus 2.
To date, no information on late-onset infection in newborns to mother with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) contracted in pregnancy are available. This study aimed to evaluate postdischarge SARS-CoV-2 status of newborns to mothers with COVID-19 in pregnancy that, at birth, were negative to SARS-CoV-2. This is an observational study of neonates born to mothers with coronavirus disease 2019 (COVID-19). Seven pregnant women with documented SARS-CoV-2 infection have been evaluated in our institution. One woman had a spontaneous abortion at 8 weeks of gestational age, four women recovered and are still in follow-up, and two women delivered. Two newborns were enrolled in the study. At birth and 3 days of life, newborns were negative to SARS-CoV-2. At 2-week follow-up, one newborn tested positive although asymptomatic. Our findings highlight the importance of follow-up of newborns to mothers with COVID-19 in pregnancy, since they remain at risk of contracting the infection in the early period of life and long-term consequences are still unknown. · Newborns to mothers with coronavirus disease 2019 (COVID-19) in pregnancy can acquire the infection later after birth.. · Newborns to mothers with COVID-19 in pregnancy need a long-term follow-up, even if they tested negative at birth.. · Specific guidelines for the long-term follow-up of newborns to mothers with COVID-19 in pregnancy are needed..


Infant Behavior Questionnaire-Revised Very Short Form: A New Factor Structure's Associations With Parenting Perceptions and Child Language Outcomes.
The Infant Behavior Questionnaire Revised-Very Short Form (IBQ-R VSF; Putnam, Helbig, Gartstein, Rothbart, & Leerkes, 2014 ) is a new publicly available measure of infant temperament measuring positive affectivity/surgency (PAS), negative emotionality (NEG), and orienting and regulatory capacity (ORC). Although the initial psychometric properties of the 3-factor model appear promising, it has not been administered to a large and diverse sample and its predictive validity has not been established. This study administered the IBQ-R VSF to a diverse sample of 5,639 mothers of infants aged between 23 and 52 weeks. Confirmatory factor analysis found that the 3-factor solution did not meet the requirement for satisfactory model fit. Exploratory factor analysis found that a 5-factor solution (PAS, NEG, Orienting Capacity, Affiliation/Regulation, and Fear) was statistically and conceptually the most parsimonious. All 5 temperament dimensions were found to relate to both mother- and partner-reported infant closeness, parenting confidence, and parenting satisfaction, and four of the dimensions (PAS, Orienting Capacity, NEG, and Fear) related to the infants' communication development. Some parental differences were also found. Together these findings suggest that the 5-factor IBQ-R VSF is a promising measure of infant temperament and is related to parenting perceptions and child language development.


[Outcomes prediction for children with Hodgkin's disease].
A prognostic score was developed in the course of treatment of 551 pediatric patients with Hodgkin's disease stage I-IV (1967-2001). It took care of such significant factors of uni-and multivariate analysis as age (11 years or more), peripheral lymph node involvement (5cm or more), systemic symptoms (B), histological pattern for lymphocytic depletion, IVB stage and/or extranodal disease (E) and biological rate of tumor process ("b"). As a result, three groups at risk were identified: favorable prognosis (65.8%: overall survival - 92%, relapse-free - 67%), medium risk (17.8%: overall survival - 67%, relapse-free 48%) and high risk (16.4%: overall survival - 38%, relapse-free 25%). Our data may be used to estimate the scope of risk-adapted therapy for Hodgkin's disease.


[GATA2 deficiency].
GATA2 is a transcription factor that is involved in the lympho-hematopoiesis. Mutations of GATA2 cause MonoMAC syndrome (monocytopenia and mycobacterial infections)/DCML deficiency (dendritic cell, monocyte, B and natural killer (NK) lymphoid deficiency), Emberger syndrome (lymphoedema with MDS), and MDS/AML. In this review, we explain the new function of GATA2, and describe the clinical phenotypes, laboratory findings, pathology, genetic anomalies and etiology.