Four cases of solitary fibrous tumour of the eye and orbit: one with sarcomatous transformation after radiotherapy and one in a 5-year-old child's eyelid.
Solitary fibrous tumour (SFT) is quite a rare neoplasm involving the eye and the orbit. It is described as showing benign behaviour in adults, but malignant cases are exceptionally reported in this location. This report describes four new cases of SFT/giant cell angiofibroma (GCA) of the eyelid and orbit, one in a 5-year-old child, and one with sarcomatous dedifferentiated transformation occurring 9 years after radiotherapy. Four cases of ocular SFT/GCA were retrieved from the database of the Pathological Anatomy Unit, University of Padova; immunohistochemistry and RT-PCR were used to identify COL1A1-PDBGF fusion gene transcripts in all cases. In case 1, late relapse 9 years later was characterised by abrupt transition into a high-grade component, associated with a non-distinctive high-grade sarcomatous area. The latter component was CD34, CD99 and Bcl2 negative and smooth muscle actin positive. Molecular characterisation showed the absence of COL1A1-PDGFB fusion transcripts in cases 1, 3 and 4, excluded diagnosis of giant cell fibroblastoma in all cases. Analysis could not be performed in case 2. An eyelid SFT/GCA in a 5-year-old child is the youngest case reported in the literature, indicating that the tumour is not exclusive to adults. The case with sarcomatous transformation, with dedifferentiated features occurring 9 years after radiotherapy, raises some questions about the choice of treatment for ocular SFT, in which excision is sometimes difficult without devastating surgery.


Effect of dolutegravir-based drug combinations on the level of medication adherence and viral load among adolescents living with HIV in resource-limited setting: a pre-post design.
The use of antiretroviral drugs among HIV-infected adolescents has been challenged with poor medication adherence, leading to an unsuppressed viral load and ultimately to drug resistance. Recently, dolutegravir has been approved for use in adolescents with HIV, but the evidence on medication adherence and viral load suppression is limited. The study determined the effect of dolutegravir-based drug regimens on the level of medication adherence and viral load among treatment-experienced adolescents. A longitudinal pre-post design study was done among adolescents living with HIV at a paediatrics clinic in Nigeria. Assessment of medication adherence and viral load testing was done before and at six months of transitioning to dolutegravir-based regimens. McNemar-chi-square was used to determine the effectiveness of the drug on adherence and viral load suppression. Multivariate logistic regression analysis was performed to determine the predictors of drug adherence and viral suppression. The mean age of the participants was 14.7 years (SD=3.2), and 53.3 % were males. The mean duration of ART use was 10.3 years (SD=3.2). Six months after transitioning to dolutegravir, there was a significant decrease in viral load (Z=-7.947, p<0.001) and an increase in medication adherence score (Z=-7.554, p<0.001), among the treatment-experienced adolescents. Viral suppression was 13 times higher among respondents with good medication adherence as compared to those with poor medication adherence (AOR=13.24, CI=3.25-53.90). Dolutegravir is effective in improving medication adherence and viral suppression among Adolescents living with HIV. Thus, the need to transition eligible adolescents to dolutegravir to sustain better treatment outcomes.


How do visual and auditory cues influence calf stress reactions at weaning?
We investigated the effects of visual and auditory information related to cow and calf stress responses at weaning. Three weaning conditions were established with physical separation only (PS), physical and visual separation (VS), and physical, visual and auditory separation (VAS). Stress responses such as vocalization and self-grooming, and changes of three maintenance behaviors (lying, walking and grazing) were recorded for experimental cows and calves in the respective conditions. Comparison of the data revealed that the cow responses differed slightly among groups. However, the peak frequency of vocalization was higher in the PS calves than in those of either VS or VAS. Self-grooming frequency was significantly different in calves and was high in the order of VAS, VS and PS calves. Furthermore, the percentage of time spent lying down was greater, and that of walking was less, in VAS calves than in either VS or PS calves. These results demonstrate that different information at weaning might strongly affect the stress responses of calves but not those of cows. Relative impacts of different types sensory information on stressors must be considered to plan animal-friendly weaning.


Treatment-Resistant Bipolar Depression: Therapeutic Trends, Challenges and Future Directions.
Bipolar disorder (BD) is a chronic mental illness impacting 1-2% of the population worldwide and causing high rates of functional impairment. Patients with BD spend most of their time in depressive episodes and up to one-third of patients do not respond to adequate doses of medications. Although no consensus exists for definition of treatment-resistant bipolar depression (TRBD), failure of symptoms improvement despite an adequate trial of two therapeutic agents is a common theme of TRBD. In this paper, we review the evidence base of therapeutic interventions, challenges, and potential future directions for TRBD. We conducted a literature search for randomized controlled trials on PubMed for the treatment of TRBD and ongoing trials for the treatment of TRBD/bipolar depression on clinicaltrials.gov. Several therapeutic agents have been investigated for TRBD. Adjunctive pramipexole and modafinil have data supporting short-term efficacy in TRBD, along with limited data for racemic intravenous ketamine. Celecoxib augmentation of escitalopram and treatment with metformin in patients with insulin resistance showed promising results. Right unilateral electroconvulsive therapy displayed statistically significant response rate and improvement, but not remission compared to pharmacotherapy. Trials for transcranial magnetic stimulation (TMS) have failed to show a significant difference from sham treatment in TRBD. Pharmacological treatments with novel mechanisms of actions like brexpiprazole and vortioxetine are being investigated following successes in unipolar depression. Modified TMS protocols such as accelerated TMS are under investigation. Innovative approaches like psychedelic-assisted psychotherapy, interleukin-2, fecal microbiota transplantation and multipotent stromal cells are being studied. Evidence on current treatment modalities for TRBD is limited with low efficacy. More research is needed for successful treatment of TRBD. Effective therapies and innovative approaches to treatment are being investigated and could show promise.


Phase II trial to evaluate gemcitabine and etoposide for locally advanced or metastatic pancreatic cancer.
Prior studies suggest that tumor cell lines harboring RAS mutations display remarkable sensitivity to gemcitabine and etoposide. In a phase II clinical trial of patients with locally advanced or metastatic pancreatic cancer, we evaluated the response rate to a combination of these drugs. Forty chemo-naïve patients with nonresectable and histologically confirmed pancreatic cancer were accrued. Patients received gemcitabine 1,000 mg/m(2) (days 1 and 8) and etoposide 80 mg/m(2) (days 8, 9, and 10; 21-day cycle). The primary end point was radiological response rate. Secondary objectives were determination of overall survival, response duration (time to progression), quality of life, toxicity, and CA 19-9 biomarker response. In 35 evaluable patients, 10 exhibited a radiological partial response and 12 had stable disease in response to treatment. Twenty patients exhibited a >20% decrease in CA 19-9 biomarker levels. Median overall survival was 6.7 months for all patients (40) and 7.2 months for evaluable patients (35). Notably, four patients survived for longer than 1 year, with two patients surviving for more than 2 years. Median time to progression for evaluable patients was 3.1 months. The median overall survival for locally advanced patients was 8.8 months and 6.75 months for metastatic patients. One-year survival was 10% for all patients and 11.4% for evaluable patients. Quality of life improved in 12 patients and remained stable in 3 of the evaluable patients. The primary dose-limiting toxicities were hematologic toxicity and fatigue. These results show that the gemcitabine and etoposide combination is generally well-tolerated and exhibits a response rate similar to other published studies.


Hyperandrogenism in polycystic ovary syndrome affects psychological well-being of adolescents.
Polycystic ovary syndrome (PCOS) is a common endocrinological disorder in reproductive age and criterion have recently been revised for adolescent age group. Research regarding effects of PCOS on psychological well-being is limited; and majority of the studies are conducted in adult patients. We aimed to examine psychological effects of PCOS in adolescents who are diagnosed using latest criterion. Cases were divided into PCOS and control groups according to their clinical, ultrasonographical and biochemical features. Beck depression inventory (BDI), State & Trait Anxiety Inventory (STAI-I/II), Self-Confidence Scale (SCS), Multidimensional Peer-Victimization Scale (MPVS) and Social Anxiety Scale for Adolescents (SAS-A) were used in psychiatric evaluation. There were no statistically significant differences between groups regarding psychiatric scale scores. However, several biochemical parameters (dehydroepiandrosterone sulfate, 17-hydroxyprogesterone) and clinical (Ferriman-Gallwey score [FGS]) of hyperandrogenism affected certain indicators of psychological well-being such as social anxiety, low self-esteem and peer victimization. In some psychiatric scales, biochemical indicators were found correlated while clinical indicators were not. Similar psychiatric scale scores between groups may indicate preliminary stages for adolescent PCOS in which endocrinological, physical and social factors have not yet reached their full potential for their effect on emergence of psychological problems; thus making this age group critical for interventions of prevention measures. In addition, while examining the effects of PCOS on psychological well-being, biochemical parameters of hyperandrogenism might be as effective as physical manifestations (FGS); and high levels of biochemical parameters of hyperandrogenism might also affect psychological state.


Nutritional assessment and fate of DNA of soybean meal from roundup ready or conventional soybeans using rats.
This study was conducted to compare the safety of soybean meal prepared from genetically modified (GM) glyphosate-tolerant (Roundup Ready; RR) soybeans and conventional soybeans. Eighty Sprague-Dawley rats (40 males and 40 females) were randomly allotted to one of four groups according to sex and body weight for a 13-week feeding experiment. The rats were fed corn-based diets containing 60% conventional soybean meal, a mixture of 30% conventional and 30% RR soybean meal, 60% or 90% RR soybean meal. All diets were adjusted to an identical nutrient level except the 90% RR diet. The two soybean meals were similar in chemical analysis and amino acid composition. During the 13-week growth trial, body weight (P < 0.05) and feed intake (P < 0.05) decreased only in rats fed with 90% RR soybean meal at the first week. No treatment-related deaths occurred during the experiment. Gross necropsy findings, haematological or urinalysis values and clinical serum parameters showed no meaningful differences between rats fed the control and RR soybean meals. A 145 bp of cp4 epsps gene specific for the GM constructs from RR soybean meal or a 407 bp of lec gene from endogenous soybean DNA could not be detected in investigated masseter muscle samples. No adverse effects of glyphosate-tolerant soybean meal on rats were seen even at levels as high as 90% of the diet.


Comparing the efficiency of open and enclosed filtration systems in environmental DNA quantification for fish and jellyfish.
Water sampling and filtration of environmental DNA (eDNA) analysis have been performed by several different methods, and each method may yield a different species composition or eDNA concentration. Here, we investigated the eDNA of seawater samples directly collected by SCUBA to compare two widely used filtration methods: open filtration with a glass filter (GF/F) and enclosed filtration (Sterivex). We referred to biomass based on visual observation data collected simultaneously to clarify the difference between organism groups. Water samples were collected at two points in the Sea of Japan in May, September and December 2018. The respective samples were filtered through GF/F and Sterivex for eDNA extraction. We quantified the eDNA concentration of five fish and two cnidarian species by quantitative polymerase chain reaction (qPCR) using species-specific primers/probe sets. A strong correlation of eDNA concentration was obtained between GF/F and Sterivex; the intercepts and slopes of the linear regression lines were slightly different in fish and jellyfish. The amount of eDNA detected using the GF/F filtration method was higher than that detected using Sterivex when the eDNA concentration was high; the opposite trend was observed when the eDNA concentration was relatively low. The concentration of eDNA correlated with visually estimated biomass; eDNA concentration per biomass in jellyfish was approximately 700 times greater than that in fish. We conclude that GF/F provides an advantage in collecting a large amount of eDNA, whereas Sterivex offers superior eDNA sensitivity. Both filtration methods are effective in estimating the spatiotemporal biomass size of target marine species.


Alcohol intoxication following muscle contraction in mice decreases muscle protein synthesis but not mTOR signal transduction.
Alcohol (ethanol [EtOH]) intoxication antagonizes stimulation of muscle protein synthesis and mammalian target of rapamycin (mTOR) signaling. However, whether the anabolic response can be reversed when alcohol is consumed after the stimulus is unknown. A single bout of electrically stimulated muscle contractions (10 sets of 6 contractions) was induced in fasted male C57BL/6 mice 2 hours prior to alcohol intoxication. EtOH was injected intraperitoneally (3 g/kg), and the gastrocnemius/plantaris muscle complex was collected 2 hours later from the stimulated and contralateral unstimulated control leg. Muscle contraction increased protein synthesis 28% in control mice, while EtOH abolished this stimulation-induced increase. Further, EtOH suppressed the rate of synthesis ~75% compared to control muscle irrespective of stimulation. This decrease was associated with impaired protein elongation as EtOH increased the phosphorylation of eEF2 Thr(56) . In contrast, stimulation-induced increases in mTOR protein complex-1 (mTORC1) (S6K1 Thr(421) /Ser(424) , S6K1 Thr(389) , rpS6 Ser(240/244) , and 4E-BP1 Thr(37/46) ) and mitogen-activated protein kinase (MAPK) (JNK Thr(183) /Tyr(185) , p38 Thr(180) /Tyr(182) , and rpS6S(235/236) ) signaling were not reversed by acute EtOH. These data suggest that EtOH-induced decreases in protein synthesis in fasted mice may be independent of mTORC1 and MAPK signaling following muscle contraction and instead due to the antagonistic actions of EtOH on mRNA translation elongation. Therefore, EtOH suppresses the contraction-induced increase in protein synthesis, and over time has the potential to prevent skeletal muscle hypertrophy induced by repeated muscle contraction.


Continuous Fixed-Bed Column Studies on Congo Red Dye Adsorption-Desorption Using Free and Immobilized Nelumbo nucifera Leaf Adsorbent.
The adsorption of Congo red (CR), an azo dye, from aqueous solution using free and immobilized agricultural waste biomass of Nelumbo nucifera (lotus) has been studied separately in a continuous fixed-bed column operation. The N. nucifera leaf powder adsorbent was immobilized in various polymeric matrices and the maximum decolorization efficiency (83.64%) of CR occurred using the polymeric matrix sodium silicate. The maximum efficacy (72.87%) of CR dye desorption was obtained using the solvent methanol. Reusability studies of free and immobilized adsorbents for the decolorization of CR dye were carried out separately in three runs in continuous mode. The % color removal and equilibrium dye uptake of the regenerated free and immobilized adsorbents decreased significantly after the first cycle. The decolorization efficiencies of CR dye adsorption were 53.66% and 43.33%; equilibrium dye uptakes were 1.179 mg g-1 and 0.783 mg g-1 in the third run of operation with free and immobilized adsorbent, respectively. The column experimental data fit very well to the Thomas and Yoon-Nelson models for the free and immobilized adsorbent with coefficients of correlation R2 ≥ 0.976 in various runs. The study concludes that free and immobilized N. nucifera can be efficiently used for the removal of CR from synthetic and industrial wastewater in a continuous flow mode. It makes a substantial contribution to the development of new biomass materials for monitoring and remediation of toxic dye-contaminated water resources.


Phosphothreonine Lyase Promotes p65 Degradation in a Mitogen-Activated Protein Kinase/Mitogen- and Stress-Activated Protein Kinase 1-Dependent Manner.
Bacterial phosphothreonine lyases have been identified to be type III secretion system (T3SS) effectors that irreversibly dephosphorylate host mitogen-activated protein kinase (MAPK) signaling to promote infection. However, the effects of phosphothreonine lyase on nuclear factor κB (NF-κB) signaling remain largely unknown. In this study, we detected significant phosphothreonine lyase-dependent p65 degradation during Edwardsiella piscicida infection in macrophages, and this degradative effect was blocked by the protease inhibitor MG132. Further analysis revealed that phosphothreonine lyase promotes the dephosphorylation and ubiquitination of p65 by inhibiting the phosphorylation of mitogen- and stress-activated protein kinase-1 (MSK1) and by inhibiting the phosphorylation of extracellular signal-related kinase 1/2 (ERK1/2), p38α, and c-Jun N-terminal kinase (JNK). Moreover, we revealed that the catalytic active site of phosphothreonine lyase plays a critical role in regulating the MAPK-MSK1-p65 signaling axis. Collectively, the mechanism described here expands our understanding of the pathogenic effector in not only regulating MAPK signaling but also regulating p65. These findings uncover a new mechanism by which pathogenic bacteria overcome host innate immunity to promote pathogenesis.


An alternatively spliced domain of the NDC1 NAD(P)H dehydrogenase gene strongly influences the expression of the ACTIN2 reference gene in Arabidopsis thaliana.
In plant respiratory chains, alternative pathways for NAD(P)H oxidation are mediated by type II NAD(P)H dehydrogenases belonging to the NDA, NDB, and NDC families. For the latter type, Arabidopsis thaliana contains a single gene, NDC1, whose functional role has not previously been analyzed in the plant. We found that A. thaliana NDC1 is alternatively spliced. Four base pairs at the 3' end of intron 5 are spliced out in NDC1-1, but retained in the NDC1-2 mRNA, which therefore contains a truncated reading frame. Both variants are conserved in dicotyledonous and monocotyledonous plants and their relative abundance varies between organs and in response to light. Three analyzed NDC1 T-DNA insertion lines all displayed an early bolting phenotype. A dramatic upregulation of ACTIN2 was characteristic of two lines containing T-DNA inserts upstream of intron 5, whereas a line with an insertion downstream of the NDC1-2 reading frame had an ACTIN2 expression level identical to the wildtype. Thus, the alternatively spliced 5' domain of NDC1 strongly influences the expression of the functionally unrelated ACTIN2, which is a common reference gene for quantitative RT-PCR. Also for other reference genes, strong expressional effects were observed when comparing various mutants and wildtypes in microarray databases.


Reconstitution of the pyridoxal 5'-phosphate (PLP) dependent enzyme serine palmitoyltransferase (SPT) with pyridoxal reveals a crucial role for the phosphate during catalysis.
The pyridoxal 5'-phosphate (PLP)-dependent enzyme serine palmitoyltransferase (SPT) is required for de novo sphingolipid biosynthesis. A previous study revealed a novel and unexpected interaction between the hydroxyl group of the l-serine substrate and the 5'-phosphate group of PLP. By using pyridoxal (PL), the dephosphorylated analogue of vitamin B6, we show here that this interaction is important for substrate specificity and optimal catalytic efficiency.


Ethical challenges and opportunities for nurses in HIV and AIDS community-based participatory research in Jamaica.
As part of a multinational program of research, we undertook a community-based participatory research project in Jamaica to strengthen nurses' engagement in HIV and AIDS policy. Three leadership hubs were purposefully convened and included small groups of people (6-10) from diverse HIV and AIDS stakeholder groups in Jamaica: frontline nurses and nurse managers in primary and secondary care settings; researchers; health care decision makers; and other community members. People living with HIV or AIDS were among the hub members. Using a relational public health ethics framework, we outline some of the ethical challenges and opportunities experienced by the research team and the leadership hubs. Data included research assistant field notes and hub progress reports. Emerging ethical concerns were associated with relational personhood, social justice, relational autonomy, relational solidarity, and sustainability of the hub activities.


EGFR-mutant lung adenocarcinomas treated first-line with the novel EGFR inhibitor, XL647, can subsequently retain moderate sensitivity to erlotinib.
EGFR-mutant lung cancers are sensitive to EGFR tyrosine kinase inhibitors (TKIs). Unfortunately, they develop resistance, often due to acquisition of a second-site mutation (T790M). Current EGFR TKIs select for T790M in preclinical models of acquired resistance. We explored whether all EGFR TKIs similarly select for the T790M mutation using data from early clinical trials and established in vitro models of acquired resistance. We analyzed the clinical characteristics of eight patients with metastatic EGFR-mutant lung adenocarcinoma who were treated first-line with XL647 and then progressed. XL647 is an ATP-competitive inhibitor of EGFR, HER2, KDR, and EPHB4. Additional molecular preclinical studies were performed to characterize resistance. Four patients displayed confirmed partial responses (PRs), three patients had unconfirmed PRs, and one patient displayed stable disease. Only one of five patients' tumor samples available for analysis after disease progression harbored the T790M mutation. Eight patients subsequently received erlotinib, with (n = 3) or without (n = 5) chemotherapy. Three of five patients treated with single-agent erlotinib derived additional benefit, staying on drug up to 9 months. EGFR-mutant PC-9 cells with acquired resistance to XL647 did not harbor the T790M mutation, displayed a distinct mRNA profile from PC-9 cells with T790M-mediated resistance, and were moderately sensitive to erlotinib in growth inhibition assays. Crystal structure analyses of XL647/EGFR T790M did not reveal a different binding mode from that of erlotinib. The findings of this exploratory study suggest that different EGFR TKIs may select for distinct mechanisms of resistance. These results raise the possibility that different EGFR TKIs could be sequentially used to improve outcomes in patients with EGFR-mutant lung cancer. Further work investigating this hypothesis is warranted.


Oncological outcomes of patients with Ewing's sarcoma: is there a difference between skeletal and extra-skeletal Ewing's sarcoma?
The aim of this study was to identify whether there was any difference in patient, tumour, treatment or outcome characteristics between patients with skeletal or extra-skeletal Ewing's sarcoma. We identified 300 patients with new primary Ewing's sarcoma diagnosed between 1980 and 2005 from the centres' local database. There were 253 (84%) with skeletal and 47 (16%) with extra-skeletal Ewing's sarcomas. Although patients with skeletal Ewing's were younger (mean age 16.8 years) than those with extra-skeletal Ewing's sarcoma (mean age 27.5 years), there was little difference between the groups in terms of tumour stage or treatment. Nearly all the patients were treated with chemotherapy and most had surgery. There was no difference in the overall survival of patients with skeletal (64%) and extra-skeletal Ewing's sarcoma (61%) (p = 0.85), and this was also the case when both groups were split by whether they had metastases or not. This large series has shown that the oncological outcomes of Ewing's sarcoma are related to tumour characteristics and patient age, and not determined by whether they arise in bone or soft tissue.


Dietary adaptation of FADS genes in Europe varied across time and geography.
Fatty acid desaturase (FADS) genes encode rate-limiting enzymes for the biosynthesis of omega-6 and omega-3 long-chain polyunsaturated fatty acids (LCPUFAs). This biosynthesis is essential for individuals subsisting on LCPUFA-poor diets (for example, plant-based). Positive selection on FADS genes has been reported in multiple populations, but its cause and pattern in Europeans remain unknown. Here we demonstrate, using ancient and modern DNA, that positive selection acted on the same FADS variants both before and after the advent of farming in Europe, but on opposite (that is, alternative) alleles. Recent selection in farmers also varied geographically, with the strongest signal in southern Europe. These varying selection patterns concur with anthropological evidence of varying diets, and with the association of farming-adaptive alleles with higher FADS1 expression and thus enhanced LCPUFA biosynthesis. Genome-wide association studies reveal that farming-adaptive alleles not only increase LCPUFAs, but also affect other lipid levels and protect against several inflammatory diseases.


Safety of Fertility-Sparing Surgery for Premenopausal Women With Sex Cord-Stromal Tumors Confined to the Ovary.
The aim of this retrospective population-based study was to investigate the oncologic safety of fertility-sparing surgery (FSS) for premenopausal women with malignant sex cord-stromal tumors (SCSTs) confined to the ovary. A cohort of women aged 18 to 49 years and diagnosed with a stage I malignant SCST between 1984 and 2013 was drawn from the National Cancer Institute's Surveillance, Epidemiology, and End Results database. Based on site-specific surgery codes, women who had FSS, defined as unilateral oophorectomy/tumor resection without hysterectomy, and definite surgery were identified. Cancer-specific survival and overall survival were evaluated after generation of Kaplan-Meier curves, whereas comparisons between the 2 groups were made with the log-rank test. A total of 255 women who met the inclusion criteria were identified; 161 (63.1%) underwent FSS whereas 94 (36.9%) had definitive surgery (bilateral salpingo-oophorectomy and hysterectomy). Median follow-up was 104 months. Cancer-specific survival (P = 0.015) but not overall survival (P = 0.76) was superior for women who had definite surgery. In this retrospective population-based cohort of premenopausal women with SCSTs confined to the ovary, FSS was associated only with a worse long-term cancer-specific survival compared with definitive surgery. Women undergoing FSS for early stage SCSTs should be extensively counseled and closely monitored.


Neuromodulatory control of a goal-directed decision.
Many cost-benefit decisions reduce to simple choices between approach or avoidance (or active disregard) to salient stimuli. Physiologically, critical factors in such decisions are modulators of the homeostatic neural networks that bias decision processes from moment to moment. For the predatory sea-slug Pleurobranchaea, serotonin (5-HT) is an intrinsic modulatory promoter of general arousal and feeding. We correlated 5-HT actions on appetitive state with its effects on the approach-avoidance decision in Pleurobranchaea. 5-HT and its precursor 5-hydroxytryptophan (5-HTP) augmented general arousal state and reduced feeding thresholds in intact animals. Moreover, 5-HT switched the turn response to chemosensory stimulation from avoidance to orienting in many animals. In isolated CNSs, bath application of 5-HT both stimulated activity in the feeding motor network and switched the fictive turn response to unilateral sensory nerve stimulation from avoidance to orienting. Previously, it was shown that increasing excitation state of the feeding network reversibly switched the turn motor network response from avoidance to orienting, and that 5-HT levels vary inversely with nutritional state. A simple model posits a critical role for 5-HT in control of the turn network response by corollary output of the feeding network. In it, 5-HT acts as an intrinsic neuromodulatory factor coupled to nutritional status and regulates approach-avoidance via the excitation state of the feeding network. Thus, the neuromodulator is a key organizing element in behavioral choice of approach or avoidance through its actions in promoting appetitive state, in large part via the homeostatic feeding network.


[Mucociliary transport and motility of the ciliary system of the nasal mucosa in patients with chronic polypous rhinosinusitis].
A transport function and motor activity of the ciliary system (MACS) of the nasal mucosa was studied in 10 patients with chronic polypous rhinosinusitis (CPRS). A complex examination included optic endoscopy of the nasal cavity for assessment of nasal polypous process, timing of the saccharine test (ST) according to standard technique, and ciliary beat rate (BR) by V.S. Kozlov method. For estimation of ciliary BR epithelium was taken from five zones: inferior and middle turbinated bones, nasal septum, base and top of the polyp growing from the median nasal passage. ST time in CPRS is much higher (28.3+/-0.34 min) than normal age-specific values. Ciliary BR in different nasal anatomic zones in CPRS is not similar. MACS on the inferior turbinated bone and nasal septum was absent, none of the samples contained cells with functioning cilia. Ciliary BR remained rather high, despite the pathological process, at the site of the top and base of the polyps as well as on the middle turbinated bone (8.7+/-1.07, 8.1+/-1.02, 6.5+/-0.35 Hz, respectively).


Core Muscle Activity during TRX Suspension Exercises with and without Kinesiology Taping in Adults with Chronic Low Back Pain: Implications for Rehabilitation.
This study aimed to examine the effects of kinesiology taping (KT) and different TRX suspension workouts on the amplitude of electromyographic (EMG) activity in the core muscles among people with chronic low back pain (LBP). Each participant (total n = 21) was exposed to two KT conditions: no taping and taping, while performing four TRX suspension exercises: (1) hamstring curl, (2) hip abduction in plank, (3) chest press, and (4) 45-degree row. Right transversus abdominis/internal oblique (TrAIO), rectus abdominis (RA), external oblique (EO), and superficial lumbar multifidus (LMF) activity was recorded with surface EMG and expressed as a percentage of the EMG amplitude recorded during a maximal voluntary isometric contraction of the respective muscles. Hip abduction in plank increased TrAIO, RA, and LMF EMG amplitude compared with other TRX positions (P < 0.008). Only the hamstring curl was effective in inducing a high EMG amplitude of LMF (P < 0.001). No significant difference in EMG magnitude was found between the taping and no taping conditions overall (P > 0.05). Hip abduction in plank most effectively activated abdominal muscles, whereas the hamstring curl most effectively activated the paraspinal muscles. Applying KT conferred no immediate benefits in improving the core muscle activation during TRX training in adults with chronic LBP.


Impact of advanced age on survival in patients with implantable cardioverter defibrillators.
Given the selectivity of clinical trial patients and meager representation of elderly in the major implantable cardioverter defibrillator (ICD) randomized trials (<25%), whether such data apply to elderly patients overall is unclear. The purpose of our study is to understand the influence of age on mortality after ICD implantation. We performed a retrospective cohort study of 502 consecutive patients receiving ICDs from 1993 to 2003 at a single university hospital. The primary predictor was patient age, categorized as <65, 65-75, and >75 years at ICD implantation. The primary outcome was time to death. Mean follow-up was 4 years. Median survival after ICD implantation was 5.3 years among subjects >75 years, less than half that of the youngest group. After adjusting for potential confounders, compared with subjects <65 years of age, patients >75 years [hazard ratio (HR), 4.7; 95% confidence interval (CI), 2.8-7.9; P < 0.001] and those 65-75 years (HR, 2.8; 95% CI, 1.7-4.8; P < 0.001) were at greater risk of death. Increased age was associated with higher total, cardiac, and non-cardiac mortality (all P <or= 0.001). Age at ICD implantation is strongly and independently associated with mortality. Age should be considered among potential co-morbidities in anticipating survival of the elderly patient prior to ICD implantation.


[Role of hormonal regulation and stress in the interaction of sleep and biorhythms in aging].
The present work describes bibliographic analysis of ontogenetic changes in hormonal regulation of sleep and circadian biorhythms. It is suggested that two-stage, age-related dynamics of such changes may be related to differential occurrence registered by us earlier, of maxima of morbidity and mortality caused by a number of disorders, in middle-age and in senile period. It is proposed to concentrate future efforts on elaboration of mathematical models of ontogenetic regulation, on the basis of systems biomedicine.


Fermion Doubling Theorems in Two-Dimensional Non-Hermitian Systems for Fermi Points and Exceptional Points.
The fermion doubling theorem plays a pivotal role in Hermitian topological materials. It states, for example, that Weyl points must come in pairs in three-dimensional semimetals. Here, we present an extension of the doubling theorem to non-Hermitian lattice Hamiltonians. We focus on two-dimensional non-Hermitian systems without any symmetry constraints, which can host two different types of topological point nodes, namely, (i) Fermi points and (ii) exceptional points. We show that these two types of protected point nodes obey doubling theorems, which require that the point nodes come in pairs. To prove the doubling theorem for exceptional points, we introduce a generalized winding number invariant, which we call the "discriminant number." Importantly, this invariant is applicable to any two-dimensional non-Hermitian Hamiltonian with exceptional points of arbitrary order and, moreover, can also be used to characterize nondefective degeneracy points. Furthermore, we show that a surface of a three-dimensional system can violate the non-Hermitian doubling theorems, which implies unusual bulk physics.


The histone demethylase jumonji coordinates cellular senescence including secretion of neural stem cell-attracting cytokines.
Jumonji domain-containing protein 3 (JMJD3/KDM6B) demethylates lysine 27 on histone H3 (H3K27me3), a repressive epigenetic mark controlling chromatin organization and cellular senescence. To better understand the functional consequences of JMJD3 its expression was investigated in brain tumor cells. Querying patient expression profile databases confirmed JMJD3 overexpression in high-grade glioma. Immunochemical staining of two glioma cell lines, U251 and U87, indicated intrinsic differences in JMJD3 expression levels that were reflected in changes in cell phenotype and variations associated with cellular senescence, including senescence-associated β-galactosidase (SA-β-gal) activity and the senescence-associated secretory phenotype (SASP). Overexpressing wild-type JMJD3 (JMJD3wt) activated SASP-associated genes, enhanced SA-β-gal activity, and induced nuclear blebbing. Conversely, overexpression of a catalytically inactive dominant negative mutant JMJD3 (JMJD3mut) increased proliferation. In addition, a large number of transcripts were identified by RNA-seq as altered in JMJD3 overexpressing cells, including cancer- and inflammation-related transcripts as defined by Ingenuity Pathway Analysis. These results suggest that expression of the SASP in the context of cancer undermines normal tissue homeostasis and contributes to tumorigenesis and tumor progression. These studies are therapeutically relevant because inflammatory cytokines have been linked to homing of neural stem cells and other stem cells to tumor loci. This glioma study brings together actions of a normal epigenetic mechanism (JMJD3 activity) with dysfunctional activation of senescence-related processes, including secretion of SASP proinflammatory cytokines and stem cell tropism toward tumors.


Implementing recommended screening for critical congenital heart disease.
Critical congenital heart disease (CCHD) is endorsed by the US Secretary of Health and Human Services as part of the recommended uniform screening panel for newborns. Although initial recommendations for implementation exist, as states and hospitals have moved forward with implementation of screening, new challenges and areas for additional focus have been identified. The objective of this study was to develop recommendations to address current challenges and areas of focus surrounding CCHD newborn screening. A workgroup of experts and stakeholders was convened in Washington, District of Columbia, for a 1-day meeting in February 2012. At the beginning of the meeting, the stakeholders held a brainstorming session to identify areas of main priority based on their experience. After this, stakeholders broke into small groups to refine recommendations, which were then finalized by consensus. Recommendations to address selection of screening equipment, standards for reporting of screening outcomes to stakeholders, training of health care providers and educating families, future research priorities, payment for screening, follow-up diagnostic testing, and public health oversight, and advocacy to facilitate effective and comprehensive screening were proposed. Suggestions for future work were developed. Screening for CCHD presents novel challenges and opportunities; however, addressing these will strengthen newborn screening and newborn care networks, and ultimately improve health outcomes.


Liver glycogen synthase and phosphorylase changes in vivo with hypoxia and anesthetics.
Methods for obtaining and processing rat liver for determination of glycogen phosphorylase a and synthase I activity were studied. An extremely rapid and profound increase in phosphorylase was induced by hypoxia. The effect on synthase I was slower and less striking. Using alpha- and beta-adrenergic antagonists, a catecholamine-depleting agent, and a ganglionic blocking agent, it was determined that adrenergic stimulation secondary to the surgical procedure required to obtain the liver was not a significant factor. The anesthetic agent used also had a significant effect on the proportion of phosphorylase in the a form. Seconal anesthesia resulted in lower phosphorylase a levels than did ether or urethan anesthesia.


Improvement in pruritus in children with atopic dermatitis using pimecrolimus cream 1%.
The objective of this study was to assess time to onset of pruritus improvement in a pediatric population treated with pimecrolimus cream 1%. This 8-day, double-blinded, vehicle-controlled study randomized 174 children and adolescents (aged 2-17 years) with mild to moderate atopic dermatitis (AD) and moderate to severe pruritus to twice-daily applications of pimecrolimus cream 1% or vehicle. There were no significant between-group differences in demographics or baseline disease characteristics. Pruritus was assessed by subjects using a 4-point pruritus severity scale (0-3). The primary efficacy variable was time to a 1 point or more improvement in pruritus score from baseline. The 2 treatment groups were compared using log-rank testing of the time-to-event data. In the per-protocol (PP) population, median times to a 1 point or more improvement in pruritus score were 48 and 72 hours for pimecrolimus and vehicle groups, respectively (P = .038). From day 3 onward, significantly more subjects (P = .023) in the pimecrolimus group versus the vehicle group reported complete pruritus resolution. Pimecrolimus cream 1% improved pruritus within 48 hours in children and adolescents with mild to moderate AD and achieved complete resolution of pruritus in a significantly greater number of subjects in the pimecrolimus group versus the vehicle group by the end of the 7-day treatment period (P = .008).


Thermal energy storage characteristics of micro-nanoencapsulated heneicosane and octacosane with poly(methylmethacrylate) shell.
In this study, PMMA/heneicosane (C21) and PMMA/octacosane (C28) micro-nano capsules were fabricated via emulsion polymerisation method. The chemical structures of the fabricated capsules were verified with the FT-IR spectroscopy analysis. The results of POM, SEM and PSD analysis indicated that most of the capsules were consisted of micro/nano-sized spheres with compact surface. The DSC measurements showed that the capsules had melting temperature in the range of about 39-60 °C and latent heat energy storage capacity in the range of about 138-152 J/g. The results of TGA showed that sublimit temperature values regarding the first degradation steps of both capsules were quite over the phase change or working temperatures of encapsulated paraffins. The thermal cycling test exhibited that the capsules had good thermal reliability and chemical stability. Additionally, the prepared capsules had reasonably high thermal conductivity.


Transcriptomics for radiation biodosimetry: progress and challenges.
Transcriptomic-based approaches are being developed to meet the needs for large-scale radiation dose and injury assessment and provide population triage following a radiological or nuclear event. This review provides background and definition of the need for new biodosimetry approaches, and summarizes the major advances in this field. It discusses some of the major model systems used in gene signature development, and highlights some of the remaining challenges, including individual variation in gene expression, potential confounding factors, and accounting for the complexity of realistic exposure scenarios. Transcriptomic approaches show great promise for both dose reconstruction and for prediction of individual radiological injury. However, further work will be needed to ensure that gene expression signatures will be robust and appropriate for their intended use in radiological or nuclear emergencies.


Microarray analysis of relative gene expression stability for selection of internal reference genes in the rhesus macaque brain.
Normalization of gene expression data refers to the comparison of expression values using reference standards that are consistent across all conditions of an experiment. In PCR studies, genes designated as "housekeeping genes" have been used as internal reference genes under the assumption that their expression is stable and independent of experimental conditions. However, verification of this assumption is rarely performed. Here we assess the use of gene microarray analysis to facilitate selection of internal reference sequences with higher expression stability across experimental conditions than can be expected using traditional selection methods.We recently demonstrated that relative gene expression from qRT-PCR data normalized using GAPDH, ALG9 and RPL13A expression values mirrored relative expression using quantile normalization in Robust Multichip Analysis (RMA) on the Affymetrix GeneChip rhesus Macaque Genome Array.Having shown that qRT-PCR and Affymetrix GeneChip data from the same hormone replacement therapy (HRT) study yielded concordant results, we used quantile-normalized gene microarray data to identify the most stably expressed among probe sets for prospective internal reference genes across three brain regions from the HRT study and an additional study of normally menstruating rhesus macaques (cycle study). Gene selection was limited to 575 previously published human "housekeeping" genes. Twelve animals were used per study, and three brain regions were analyzed from each animal. Gene expression stabilities were determined using geNorm, NormFinder and BestKeeper software packages. Sequences co-annotated for ribosomal protein S27a (RPS27A), and ubiquitin were among the most stably expressed under all conditions and selection criteria used for both studies. Higher annotation quality on the human GeneChip facilitated more targeted analysis than could be accomplished using the rhesus GeneChip. In the cycle study, multiple probe sets annotated for actin, gamma 1 (ACTG1) showed high signal intensity and were among the most stably expressed. Using gene microarray analysis, we identified genes showing high expression stability under various sex-steroid environments in different regions of the rhesus macaque brain. Use of quantile-normalized microarray gene expression values represents an improvement over traditional methods of selecting internal reference genes for PCR analysis.


Hormonal regulation of collagenolysis in uterine cervical fibroblasts. Modulation of synthesis of procollagenase, prostromelysin and tissue inhibitor of metalloproteinases (TIMP) by progesterone and oestradiol-17 beta.
Rabbit uterine cervical fibroblasts produced a large amount of matrix metalloproteinases (MMPs) such as collagenase (MMP-1) and stromelysin (MMP-3) and a small relatively amount of tissue inhibitor of metalloproteinases (TIMP). When cells were treated with progesterone or oestradiol-17 beta, both steroids concurrently decreased the level of procollagenase and prostromelysin in the culture media and the steady-state levels of the respective mRNAs. On the other hand, the level of TIMP in the culture media and the steady-state level of its mRNA were simultaneously increased by these steroids. Similarly, the suppression of production of MMPs and the augmentation of TIMP production by both steroids were observed with interleukin 1 (IL-1)-treated cells, but the action of progesterone was more effective than that of oestradiol-17 beta in the IL-1-untreated and -treated cells. These results suggest that collagenolysis in uterine cervical fibroblasts is negatively regulated by steroid hormones via the acceleration of TIMP production and the suppression of synthesis of MMPs at the pretranslational level.


CCI-779 inhibits cell-cycle G2-M progression and invasion of castration-resistant prostate cancer via attenuation of UBE2C transcription and mRNA stability.
The cell-cycle G(2)-M phase gene UBE2C is overexpressed in various solid tumors including castration-resistant prostate cancer (CRPC). Our recent studies found UBE2C to be a CRPC-specific androgen receptor (AR) target gene that is necessary for CRPC growth, providing a potential novel target for therapeutic intervention. In this study, we showed that the G(1)-S cell-cycle inhibitor-779 (CCI-779), an mTOR inhibitor, inhibited UBE2C mRNA and protein expression in AR-positive CRPC cell models abl and C4-2B. Treatment with CCI-779 significantly decreased abl cell proliferation in vitro and in vivo through inhibition of cell-cycle progression of both G(2)-M and G(1)-S phases. In addition, exposure of abl and C4-2B cells to CCI-779 also decreased UBE2C-dependent cell invasion. The molecular mechanisms for CCI-779 inhibition of UBE2C gene expression involved a decreased binding of AR coactivators SRC1, SRC3, p300, and MED1 to the UBE2C enhancers, leading to a reduction in RNA polymerase II loading to the UBE2C promoter, and attenuation of UBE2C mRNA stability. Our data suggest that, in addition to its ability to block cell-cycle G(1) to S-phase transition, CCI-779 causes a cell-cycle G(2)-M accumulation and an inhibition of cell invasion through a novel UBE2C-dependent mechanism, which contributes to antitumor activities of CCI-779 in UBE2C overexpressed AR-positive CRPC.


The Long-Term Outcomes of Endoscopic Papillectomy and Management of Cases of Incomplete Resection: A Single-Center Study.
Endoscopic papillectomy is increasingly performed as an alternative to surgery for early ampullary tumors. This retrospective study aimed to evaluate the long-term results of endoscopic papillectomy, the management of cases with incomplete endoscopic resection, and the long-term recurrence rates. All 46 patients who underwent endoscopic papillectomy for ampullary tumors between November 2003 and March 2018 were retrospectively evaluated. The final pathological diagnoses were adenoma (n = 44) and adenocarcinoma (n = 2). Histopathological evaluations after endoscopic papillectomy revealed that complete resection was achieved in 19 patients (19/46, 41.3%). Among the 27 patients with incomplete resection, the margin was histopathologically positive in 14 patients and difficult to evaluate in 13. Additional surgery was performed for 2 of the 14 patients with positive margins. Excluding 2 patients who received additional surgery, 7 of the 25 patients with incomplete resection had recurrence, and 18 had no recurrence during the follow-up period. Ten (77%) of the 13 patients in whom the margin was difficult to evaluate had no recurrence. Approximately 80% of the patients in whom the histopathological evaluation of the resected margin was difficult had no recurrence even after approximately 5 years of follow-up. Thus, careful observation may be considered for these patients.


Stachydrine ameliorates pressure overload-induced diastolic heart failure by suppressing myocardial fibrosis.
Stachydrine (Sta), a major constituent of Leonurus japonicus Houtt, has been reported to possess numerous cardioprotective effects. In this study, we evaluated the effect of Sta on pressure overload-induced diastolic heart failure in rats and investigated the mechanisms underlying the effect. Wistar rats were randomized to transverse aortic constriction (TAC) or sham operation. After 3 days, the rats that underwent TAC were randomized to treatment for a total of four experimental groups (n=10 each group): sham operation, TAC only, TAC + telmisartan (Tel), and TAC + stachydrine (Sta). After 12 weeks, we evaluated left ventricular hypertrophy, function, and fibrosis by echocardiography, pressure-volume loop analysis, and histology. In addition, levels of fibrosis-related proteins in the heart were determined by Western blot analysis. Our results showed that Sta significantly suppressed TAC-induced cardiac hypertrophy, and TAC-induced increases in heart weight/body weight and heart weight/tibial length. In addition, Sta attenuated TAC-induced decreases in left ventricular ejection fraction and improved other hemodynamic parameters. Compared with the TAC only group, rats treated with Sta exhibited significant decreases in interstitial and perivascular fibrosis, TGF-βR1 protein levels, and phosphorylation of Smad2/3; however, protein levels of TGF-β1, TGF-βR2, and Smad4 did not differ significantly between the two groups. Taken together, our results demonstrate that Sta protects against diastolic heart failure by attenuating myocardial hypertrophy and fibrosis via the TGF-β/Smad pathway.


Utilization of telemedicine to support caregivers of young children with ASD and their Part C service providers: a comparison of intervention outcomes across three models of service delivery.
Families of young children with autism spectrum disorder (ASD) frequently experience barriers to accessing evidence-based early intervention services. Telemedicine presents an opportunity to increase access to these services, particularly for families in rural and under-resourced areas. The present article describes a brief behavioral intervention and support model for families of young children with concerns for ASD. In the context of the COVID-19 pandemic, this service model shifted to telemedicine-only service delivery, resulting in an opportunity to analyze intervention outcomes from services delivered either via traditional in-person visits, telemedicine-only sessions, or a hybrid model including both in-person and telemedicine sessions. Data are presented for 115 families with toddlers 16-33 months of age who participated in a six-session behavioral intervention and support service model either in-person, through telemedicine, or through a hybrid service model. This intervention was available for families referred for ASD evaluation through the state Part C early intervention program. Intervention feasibility, fidelity of implementation, child outcomes, and stakeholder satisfaction are compared across service delivery models. Caregivers, behavioral consultants, and Part C early intervention providers reported satisfaction with services, regardless of service delivery model. Caregivers and consultants also reported positive child outcomes. Statistically significant differences emerged for caregiver- and consultant-reported child outcomes in some domains, with stakeholders in the telemedicine-only group reporting slightly less improvement, compared to stakeholders in the in-person-only group. Caregivers and consultants in the telemedicine-only group also provided qualitative feedback on benefits and challenges related to telemedicine services. Both caregivers and behavioral consultants reported positive outcomes following a brief behavioral intervention and support model targeted at families of young children with concern for ASD. Stakeholders reported improvement in child behavior and satisfaction with services across in-person, telemedicine-only, and hybrid models of service delivery. These results suggest that telemedicine presents a promising opportunity for increasing service access. Additional research is needed to continue optimizing the experience of telemedicine-based service delivery for both families and intervention providers.


Evidence for frequency-dependent cortical plasticity in the human brain.
Frequency-dependent plasticity (FDP) describes adaptation at the synapse in response to stimulation at different frequencies. Its consequence on the structure and function of cortical networks is unknown. We tested whether cortical "resonance," favorable stimulation frequencies at which the sensory cortices respond maximally, influenced the impact of FDP on perception, functional topography, and connectivity of the primary somatosensory cortex using psychophysics and functional imaging (fMRI). We costimulated two digits on the hand synchronously at, above, or below the resonance frequency of the somatosensory cortex, and tested subjects' accuracy and speed on tactile localization before and after costimulation. More errors and slower response times followed costimulation at above- or below-resonance, respectively. Response times were faster after at-resonance costimulation. In the fMRI, the cortical representations of the two digits costimulated above-resonance shifted closer, potentially accounting for the poorer performance. Costimulation at-resonance did not shift the digit regions, but increased the functional coupling between them, potentially accounting for the improved response time. To relate these results to synaptic plasticity, we simulated a network of oscillators incorporating Hebbian learning. Two neighboring patches embedded in a cortical sheet, mimicking the two digit regions, were costimulated at different frequencies. Network activation outside the stimulated patches was greatest at above-resonance frequencies, reproducing the spread of digit representations seen with fMRI. Connection strengths within the patches increased following at-resonance costimulation, reproducing the increased fMRI connectivity. We show that FDP extends to the cortical level and is influenced by cortical resonance.


Mechanisms of beta-adrenergic receptor regulation in cultured chick heart cells. Role of cytoskeleton function and protein synthesis.
To examine mechanisms by which cardiac tissue regulates the beta-adrenergic receptor and physiological response to beta-adrenergic agonists, we studied the effects of cytoskeletal disrupting agents and inhibition of protein synthesis on receptor properties and contractile response to isoproterenol in intact cultured ventricular cells from embryonic chick heart. Thirty minutes of exposure of intact cells to 1 microM isoproterenol produced loss of the high-affinity state (KD = 4.5 +/- 1.5 nM) of the receptor found in cell membranes with no loss of total receptor number, whereas there was concomitant decline in the contractile response to 1 microM isoproterenol to 41 +/- 16% (SD) of control. Contractile response recovered within 60 minutes of agonist removal to 78 +/- 11% of initial response. There was concomitant recovery of the high-affinity state of the receptor, so that 1 hour after agonist removal there was 72% of the initial proportion of high-affinity receptors. This desensitization of the contractile response, as well as recovery after agonist removal, was markedly blunted by preincubation with cytochalasin B so that contractile responsiveness to isoproterenol was maintained at 77 +/- 13% of the initial response. Colchicine (10 microM) was without effect on the first 30 minutes of agonist-induced desensitization. More prolonged agonist exposure (1 microM isoproterenol for 24 hours) produced colchicine-sensitive loss of receptors from intact cells to 40% of control levels. Full recovery of receptor number occurred over 72 hours; this was completely blocked by cycloheximide (P less than 0.01). Thus, rapid desensitization and resensitization of the beta-receptor-mediated contractile response is associated with alterations in high-affinity agonist binding and appears to be modulated by microfilaments. Receptor down-regulation is dependent on functional microtubules, and recovery of these receptors after agonist removal requires protein synthesis.


Spectral ballistic imaging: a novel technique for viewing through turbid or obstructing media.
We propose a new method for viewing through turbid or obstructing media. The medium is illuminated with a modulated cw laser and the amplitude and phase of the transmitted (or reflected) signal is measured. This process takes place for a set of wavelengths in a certain wide band. In this way we acquire the Fourier transform of the temporal output. With this information we can reconstruct the temporal shape of the transmitted signal by computing the inverse transform. The proposed method benefits from the advantages of the first-light technique: high resolution, simple algorithms, insensitivity to boundary condition, etc., without suffering from its main deficiencies: complex and expensive equipment.


Clinical application of perfusion computed tomography in neurosurgery.
Currently, perfusion CT (PCT) is a valuable imaging technique that has been successfully applied to the clinical management of patients with ischemic stroke and aneurysmal subarachnoid hemorrhage (SAH). However, recent literature and the authors' experience have shown that PCT has many more important clinical applications in a variety of neurosurgical conditions. Therefore, the authors share their experiences of its application in various diseases of the cerebrovascular, neurotraumatology, and neurooncology fields and review the pertinent literature regarding expanding PCT applications for neurosurgical conditions, including pitfalls and future developments. A pertinent literature search was conducted of English-language articles describing original research, case series, and case reports from 1990 to 2011 involving PCT and with relevance and applicability to neurosurgical disorders. In the cerebrovascular field, PCT is already in use as a diagnostic tool for patients suspected of having an ischemic stroke. Perfusion CT can be used to identify and define the extent of the infarct core and ischemic penumbra core, and thus aid patient selection for acute reperfusion therapy. For patients with aneurysmal SAH, PCT provides assessment of early brain injury, cerebral ischemia, and infarction, in addition to vasospasm. It may also be used to aid case selection for aggressive treatment of patients with poor SAH grade. In terms of oncological applications, PCT can be used as an imaging biomarker to assess angiogenesis and response to antiangiogenetic treatments, differentiate between glioma grades, and distinguish recurrent tumor from radiation necrosis. In the setting of traumatic brain injury, PCT can detect and delineate contusions at an early stage. In patients with mild head injury, PCT results have been shown to correlate with the severity and duration of postconcussion syndrome. In patients with moderate or severe head injury, PCT results have been shown to correlate with patients' functional outcome. Perfusion CT provides quantitative and qualitative data that can add diagnostic and prognostic value in a number of neurosurgical disorders, and also help with clinical decision making. With emerging new technical developments in PCT, such as characterization of blood-brain barrier permeability and whole-brain PCT, this technique is expected to provide more and more insight into the pathophysiology of many neurosurgical conditions.


The physiological responses of chronic heart failure patients to maximal strength test and a balke incremental test.
It has been demonstrated that resistance exercises may improve chronic heart failure (CHF) patients' functional ability and quality of life, however, physicians do not recommend this form of exercise because of a concern for reported increases in afterload and blood pressure (BP) during the exercise. This study compared the heart rate (HR), BP and rate pressure product (RPP) of CHF patients for a Balke incremental test and a maximal strength test (MS). Fifteen men diagnosed with CHF participated in the study. All subjects performed both a Balke incremental test and MS test for eight different resistance exercises. The subjects' HR and BP were monitored during the incremental test and immediately after each resistance exercise. HR, systolic BP and RPP were significantly lower during the MS test than during both the peak Balke incremental test and during exercise at 80% of peak VO2 (p < 0.05). No significant RPP differences were found between upper and lower body resistance exercises (p > 0.05). The physiological responses in this study were less severe during a MS test than those reported during an incremental Balke treadmill test. Also the finding suggests that MS tests may be an acceptable method to assess the maximal strength of patients with moderate heart failure. Key PointsThe physiological responses of CHF patients to maximal strength test were less severe than those reported during a walking incremental test.There were similar hemodynamic responses during upper and lower resistance exercises.Maximal strength test appears to be an acceptable method to assess the maximal strength of patients with moderate CHF.


Successful rituximab therapy in refractory autoimmune hepatitis and Evans syndrome.
A 44-year-old woman was found to have elevated aminotransferases, twice the upper limit of normal. Liver biopsy demonstrated a mixed inflammatory process suggestive of both primary biliary cirrhosis and autoimmune hepatitis (AIH). Prednisone and azathioprine were started, with normalization of aminotransferases. Six months later, she returned with worsening pruritus and re-evaluation demonstrated probable reactivation of AIH with acute elevation of liver injury tests. Repeat liver biopsy was suggestive of a flare of AIH which did not respond to prednisone, azathioprine, or mycophenolate mofetil. One month later the patient was hospitalized for sudden onset of anemia and thrombocytopenia, suggestive of autoimmune hemolytic anemia and idiopathic thrombocytopenic purpura consistent with Evans syndrome. Rituximab was initiated and mycophenolate mofetil discontinued. After one infusion of rituximab, liver injury tests significantly improved. Within four weeks of rituximab infusion (4 doses) the patient's Evans syndrome completely resolved with normal hemoglobin and platelet levels; aminotransferases also significantly improved to less than twice the upper limit of normal.


Effect of chronic pain on fentanyl self-administration in mice.
The development of opioid addiction in subjects with established chronic pain is an area that is poorly understood. It is critically important to clearly understand the neurobiology associated with propensity toward conversion to addiction under conditions of chronic pain. To pose the question whether the presence of chronic pain influences motivation to self-administer opioids for reward, we applied a combination of rodent models of chronic mechanical hyperalgesia and opioid self-administration. We studied fentanyl self-administration in mice under three conditions that induce chronic mechanical hyperalgesia: inflammation, peripheral nerve injury, and repeated chemotherapeutic injections. Responding for fentanyl was compared among these conditions and their respective standard controls (naïve condition, vehicle injection or sham surgery). Acquisition of fentanyl self-administration behavior was reduced or absent in all three conditions of chronic hyperalgesia relative to control mice with normal sensory thresholds. To control for potential impairment in ability to learn the lever-pressing behavior or perform the associated motor tasks, all three groups were evaluated for acquisition of food-maintained responding. In contrast to the opioid, chronic hyperalgesia did not interfere with the reinforcing effect of food. These studies indicate that the establishment of chronic hyperalgesia is associated with reduced or ablated motivation to seek opioid reward in mice.


A retrospective study of complications of total thyroidectomy; is it a safe approach for benign thyroid conditions.
To assess frequency of complications of total thyroidectomy for benign thyroid diseases. The retrospective descriptive study was conducted at Shifa College of Medicine, Islamabad, Pakistan, and comprised patient data from March 2016 to March 2018 of all patients who had undergone total thyroidectomy at the centre. Operative findings and post-op complications were documented in a self-generated proforma. Followup visits documented in the charts were also reviewed. Patients who did not keep the follow-up appointment were interviewed over the phone. Data was analysed using SPSS 20. Of the 96 patients, 78(81.2%) were female and 18(18.7%) were male. The overall mean age was 32.9 } 9.84 years. Hypocalcaemia was the most common complication in 4(4.1%) cases, transient in 3(3.1%) and permanent in only 1(1.04%) case. Haemorrhage occurred in 1(1.04) patient, and recurrent laryngeal nerve injury in 2(2.08%). . Simple multi-nodular goitre was effectively treated with total thyroidectomy with minor complications.


The veno-occlusive disease of the liver.
The veno-occlusive disease of the liver (VOD) is a disorder caused by the non-thrombotic occlusion of the central veins of hepatic lobules. The clinical features are similar to those of intrahepatic portal hypertension (unexplained weight gain, ascites, painful hepatomegaly, jaundice). In the past, this disease was rather infrequent and was linked to the absorption of toxic agents, liver irradiation or chemotherapy. However, the intensification of treatment protocols before hematopoietic stem cell transplants has considerably increased its incidence. The strategies used for its prevention and treatment remain limited in efficacy. The present review was undertaken in order to assess progress in the diagnosis and management of this severe complication in stem cell transplantation. The method used for preparing this review was an examination of 250 relevant articles or abstracts published in journals covered by Medline. Despite the progress made toward the understanding of its physiopathology and the identification of its risk factors, VOD is still one of the leading causes of morbidity and mortality during the first two months post-BMT, and therefore often constitutes a limitation for the further increment of the dose of antineoplastic drugs. This may be explained by the difficulty in making an early diagnosis of this problem, at a time when therapeutic intervention may be more effective, and, on the other hand, the lack of a well-established prevention and treatment approach for patients with VOD. New diagnostic procedures, such as laparoscopic liver biopsy, and new therapeutic approaches, such as transjugular intrahepatic portosystemic shunting (TIPS) or defibrotide, are now being evaluated. However, additional studies will be needed to determine the most appropriate therapy for each VOD patient depending on the severity of the disease.


Thermomechanical modification of diffraction gratings.
The most accurate approaches to fabrication of diffraction gratings are known to be the lithographic and holographic methods. The lithographic methods allow fabrication of arbitrarily chirped gratings whose performance, however, is degraded by stitching errors. The holographic methods are free from stitching errors; however, they are limited in the achievable spatial variations of their grating periods. We suggest a method of diffraction grating modification by nonuniform heating and stretching that is much more flexible than the holographic approach and does not suffer from the problem of stitching error. We demonstrate our approach for quartz phase masks that have a characteristic grating period of 1 microm and a length of several centimeters. Our approach allows the grating periods of the phase masks to vary in a range from a few picometers to a few nanometers and a spatial resolution of a few millimeters. It is shown that the grating period can be modified with a negligible effect on the profile of the gratings.


Wound Healing Activity of Carallia brachiata Bark.
The stem bark of Carallia brachiata was studied for wound healing activity. The bark was extracted with petroleum ether, ethyl acetate and methanol successively. All the extracts were screened for wound healing activity by excision and incision models in Wistar rats. The ethyl acetate and methanol extracts were found to possess significant wound healing activity. The extracts revealed the presence of sterols or triterpenoids, flavonoids, phenols, tannins, carbohydrates, fixed oils and fats.


Occupation: purposefulness and meaningfulness as therapeutic mechanisms. 1995 Eleanor Clarke Slagle Lecture.
The model of occupational functioning leads the author to conceptualize occupation both as treatment end goal and as means to remediate impairments. In both dimensions, meaningfulness and purposefulness are key therapeutic qualities. Purposefulness is hypothesized to organize behavior and meaningfulness to motivate performance. These aspects of occupation need explication through research. Some questions that need to be answered include the following: How does purposefulness of occupation-as-end organize a person's life? Does purposefulness of occupation-as-means organize particular behavioral responses? What effect does meaningfulness of occupation-as-end have on achievement of role performance? Does meaningfulness of occupation-as-means motivate perseverance and effort during therapy?


Remdesivir and SARS-CoV-2: Structural requirements at both nsp12 RdRp and nsp14 Exonuclease active-sites.
The rapid global emergence of SARS-CoV-2 has been the cause of significant health concern, highlighting the immediate need for antivirals. Viral RNA-dependent RNA polymerases (RdRp) play essential roles in viral RNA synthesis, and thus remains the target of choice for the prophylactic or curative treatment of several viral diseases, due to high sequence and structural conservation. To date, the most promising broad-spectrum class of viral RdRp inhibitors are nucleoside analogues (NAs), with over 25 approved for the treatment of several medically important viral diseases. However, Coronaviruses stand out as a particularly challenging case for NA drug design due to the presence of an exonuclease (ExoN) domain capable of excising incorporated NAs and thus providing resistance to many of these available antivirals. Here we use the available structures of the SARS-CoV RdRp and ExoN proteins, as well as Lassa virus N exonuclease to derive models of catalytically competent SARS-CoV-2 enzymes. We then map a promising NA candidate, GS-441524 (the active metabolite of Remdesivir) to the nucleoside active site of both proteins, identifying the residues important for nucleotide recognition, discrimination, and excision. Interestingly, GS-441524 addresses both enzyme active sites in a manner consistent with significant incorporation, delayed chain termination, and altered excision due to the ribose 1'-CN group, which may account for the increased antiviral effect compared to other available analogues. Additionally, we propose structural and function implications of two previously identified RdRp resistance mutations in relation to resistance against Remdesivir. This study highlights the importance of considering the balance between incorporation and excision properties of NAs between the RdRp and ExoN.


Demographic Characteristics and Husbandry and Biosecurity Practices of Small Poultry Flocks in Ontario, Canada.
As part of a 2 yr disease surveillance project of small poultry flocks, owners of birds submitted for postmortem examination to the Animal Health Laboratory were asked to complete a questionnaire designed to gather information on the characteristics of the flock and its environment, how the flock was managed, and biosecurity measures used. A total of 153 unique questionnaires were received. Personal consumption of meat or eggs was the most common reason for owning a small flock (69.3%). Almost all owners (97.4%) reported having chickens on their property, while 21.6% had waterfowl, 15.7% had turkeys, and 15.7% had game birds. Nearly 70% (69.9%) of the flocks had some degree of outdoor access. For those with indoor access, the most common bedding material provided was soft wood shavings (70.2%). Kitchen waste or leftovers were offered to 65.3% of flocks, and well water was the most common source of drinking water (80.6%). For flocks with indoor access, dedicated shoes and clothes were used when entering or cleaning the coop by fewer than half of owners, and shoes were rarely disinfected before or after contact with the flock. Most owners (93.8%) reported washing their hands after contact with their birds, although only 48.3% reported washing their hands before contact. Among owners who sourced birds from a hatchery, only 36.8% indicated that the birds had been vaccinated, and 21.1% were unsure if vaccines had been administered. Among owners using medication (60.5%), the use of antibiotics was common (60.9%). Overall, questionnaire responses describe a wide range of husbandry and biosecurity practices, often suboptimal, and point out the need for educational material for Ontario small flock owners.


Chemoradiation Vs Radical Cystectomy for Muscle-invasive Bladder Cancer: A Propensity Score-weighted Comparative Analysis Using the National Cancer Database.
To address the overarching question whether chemoradiation therapy (CMT) offers overall survival (OS) similar to that of radical cystectomy (RC) in muscle-invasive bladder cancer (MIBC), we performed analyses using the National Cancer Database. Patients diagnosed with MIBC in 2004-2014 who underwent RC or received primary CMT were identified in the National Cancer Database. Survival was estimated using the weighted Kaplan-Meier method, and propensity score-weighted Cox proportional hazards model were used to evaluate association of clinicopathologic features with outcome. Of 484,367 patients with a diagnosis of bladder cancer, 35,856 underwent RC and 4050 received CMT. After applying the exclusion/inclusion criteria, data for 15,854 patients who underwent RC and 2083 who received CMT were available for analysis. Five-year OS was 40.4% in the RC group and 29.4% in the CMT group (P <.001). OS was significantly shorter in the CMT group than in the RC group in both multivariate analysis (hazards ratio [HR] 1.15, 95% CI 1.08-1.22; P <.001) and propensity score-weighted analysis (HR 1.18, 95% CI 1.07-1.30; P <.001). Interaction terms indicated better survival after RC in patients younger than 70 years (HR 1.61, 95% CI 1.34-1.93; P <.001); subgroup analyses identified a survival benefit in patients with N0/N1 disease who underwent RC (HR 1.21, 95% CI 1.09-1.33; P <.001). OS after 1 year of treatment was increased in RC group compared to CMT group in patient with MIBC. Further studies are required to identify optimal treatment for specific patients.


Studies on methanol dehydrogenase from Hyphomicrobium X. Isolation of an oxidized form of the enzyme.
1. Double-reciprocal plots of initial reaction rates of methanol dehydrogenase [alcohol--(acceptor) oxidoreductase, EC 22.214.171.124] in vitro show patterns of parallel lines. The results with various methanol, ammonia and phenazine methosulphate concentrations can be described by an equation valid for a Ping Pong kinetic mechanism with three reactants. 2. The overall maximum velocity was the same for several primary alcohols, C(2)-deuterated ethanols and different electron acceptors, but it was significantly lower for C(1)-deuterated substrates. 3. Oxidation of the isolated enzyme with electron acceptors required the presence of ammonia and a high pH. The inclusion of cyanide or hydroxylamine during the incubation was essential to prevent enzyme inactivation. The absorbance spectrum of an oxidized form of the enzyme was clearly different from that of the isolated enzyme and the free radical was no longer present. On addition of substrate, the original absorption spectrum and electron-spin-resonance signal reappeared and a concomitant substrate oxidation was found. This reaction could be carried out at pH 7 and ammonia was not required. 4. Based on the activity of the enzyme with one-electron acceptors, the presence of a free radical and the kinetic behaviour, an oxidation of the enzyme via one-electron steps is proposed.


Posterior malleolar fracture morphology determines outcome in rotational type ankle fractures.
Rotational type ankle fractures with a concomitant fracture of the posterior malleolus are associated with a poorer clinical outcome as compared to ankle fractures without. However, clinical implications of posterior malleolar (PM) fracture morphology and pattern have yet to be established. Many studies on this subject report on fragment size, rather than fracture morphology based on computed tomography (CT). The overall purpose of the current study was to elucidate the correlation of PM fracture morphology and functional outcome, assessed with CT imaging and not with -unreliable- plain radiographs. Between January 2010 and May 2014, 194 patients with an operatively (ORIF) treated ankle fracture, were prospectively included in the randomized clinical EF3X-trial at our Level-I trauma center. The current study retrospectively included 73 patients with rotational type ankle fractures and concomitant fractures of the posterior malleolus. According to the CT-based Haraguchi fracture morphology, all patients were divided into three groups: 20 Type I (large posterolateral-oblique), 21 Type II (transverse medial-extension) and 32 Type III (small-shell fragment). At 12 weeks, 1 year and 2 years postoperatively the Foot and Ankle Outcome Scores (FAOS) and SF-36 scores were obtained, with the FAOS domain scores at two years postoperative as primary study outcome. Statistical analysis included a multivariate regression and secondary a mixed model analysis. Haraguchi Type II PM ankle fractures demonstrated significantly poorer outcome scores at two years follow-up compared to Haraguchi Types I and III. Mean FAOS domain scores at two years follow-up showed to be significantly worse in Haraguchi Type II as compared to Type III, respectively: Symptoms 48.2 versus 61.7 (p = 0.03), Pain 58.5 versus 84.4 (p < 0.01), Activities of Daily Living (ADL) 64.1 versus 90.5 (p < 0.01). Posterior malleolar ankle fractures with medial extension of the fracture line (i.e. Haraguchi Type II) are associated with significantly poorer functional outcomes. The current dogma to fix PM fractures that involve at least 25-33% of the tibial plafond may be challenged, as posterior malleolar fracture pattern and morphology - rather than fragment size - seem to determine outcome.


Constitutive expression of Heregulin induces apoptosis in an erbB-2 overexpressing breast cancer cell line SKBr-3.
We have previously reported that Heregulin (HRG)/neu differentiation factor (NDF) induced growth arrest and cellular differentiation in breast cancer cells overexpressing erbB-2 receptor. To elucidate the cellular mechanisms underlying the growth inhibition by HRG, we developed an in vitro model by transfection of HRG cDNA into the erbB-2 overexpressing breast cancer cell line, SKBr-3. We showed that the enforced expression of HRG in SKBr-3 cells induces dramatic morphological changes and pronounced inhibition of anchorage-dependent and -independent growth. Most SKBr-3/HRG-transfected (SK/HRG) cells exhibited about 15-fold increase in size and persisted as multinucleated cells with extended flattened vacuole-filled cytoplasm with reduced cell attachment. The growth suppression of SK/HRG cells was accompanied by a reduction in S phase, the presence of a G2-M cell cycle delay, and an increase in DNA aneuploid components. In addition, DNA fragmentation assays showed that HRG induced apoptosis of SKBr-3 cells. In contrast, while HRG treatment of other erbB-2 overexpressing breast cancer cell lines led to growth arrest and cell detachment, it did not induce apoptotic features. Thus, this study demonstrates that while growth arrest and cell detachment are general effects of HRG towards erbB-2 overexpressing cells, the ability of HRG to induce apoptosis is a phenomenon confined to selective cells including SKBr-3 cells.


[Role of arteriolar heterogeneity on the development of the organ response of the vasculature of the skeletal muscles of the rat to constrictor stimuli].
A correlation was found between the initial relative thickness of vascular wall and the extent of vasoconstriction in response to standard stimulation of sympathetic strings. The distribution of vessels within the wall was comprised in mathematical model which estimated interrelationships among three variables: pressure perfusate expenditure and a conventional dose of vasoconstricting agent. The model revealed that intense vasoconstrictory responses entailed a decrease in the number of functioning vessels. When perfusing the rat hindlimbs with a constant expenditure in maximal vasoconstriction, the number of functioning microvessels decreased by 46 +/- 11.9%.


Intracranial venous hypertension and the effects of venous outflow obstruction in a rat model of arteriovenous fistula.
A model of rat arteriovenous fistula (AVF) was created using a proximal common carotid artery to distal external jugular vein anastomosis. Anatomical dissections revealed that the external jugular vein is the primary vessel draining intracranial venous blood. Physiological measurements were made with the AVF open and closed, and during venous outflow occlusion of the contralateral external jugular vein. Opening the AVF increased torcular pressure from 6.5 +/- 0.6 to 13.5 +/- 1.1 mm Hg and decreased mean arterial pressure from 82.7 +/- 1.8 to 62.8 +/- 1.8 mm Hg (both P less than .05), decreasing cerebral perfusion pressure from 76.2 +/- 1.7 to 49.3 +/- 2.2 mm Hg (P less than .05). Middle cerebral artery blood flow velocity (MCA BFV) decreased from 6.8 +/- 1.1 to 4.2 +/- 0.7 cm/s (P less than 0.05). In rats with an AVF, occlusion of venous outflow increased torcular pressure to 34.8 +/- 3.1 mm Hg (P less than 0.05), MCA BFV decreased to 1.8 +/- 0.5 cm/s (P less than 0.05), and severe ischemic changes were seen on the electroencephalogram. Under this condition, torcular pressure and systemic arterial pressure had a positive linear relationship (P less than 0.05), whereas in control rats torcular pressure and arterial pressure had no relationship. Restoration of cerebral perfusion pressure by release of venous outflow occlusion and AVF closure transiently increased MCA BFV to 69% above baseline (P less than 0.05). Histological examination 1 week after permanent venous outflow occlusion revealed venous infarction, subarachnoid hemorrhage, and severe brain edema in rats with an AVF but not in control rats without an AVF. This model of cerebrovascular steal with venous hypertension reproduces both hemodynamic and hemorrhagic complications of human AVF and emphasizes the importance of venous outflow obstruction and venous hypertension in the pathophysiology of these lesions.


Alk3 controls nephron number and androgen production via lineage-specific effects in intermediate mesoderm.
The mammalian kidney and male reproductive system are both derived from the intermediate mesoderm. The spatial and temporal expression of bone morphogenetic protein (BMP) 2 and BMP4 and their cognate receptor, activin like kinase 3 (ALK3), suggests a functional role for BMP-ALK3 signaling during formation of intermediate mesoderm-derivative organs. Here, we define cell autonomous functions for Alk3 in the kidney and male gonad in mice with CRE-mediated Alk3 inactivation targeted to intermediate mesoderm progenitors (Alk3(IMP null)). Alk3-deficient mice exhibit simple renal hypoplasia characterized by decreases in both kidney size and nephron number but normal tissue architecture. These defects are preceded by a decreased contribution of Alk3-deleted cells to the metanephric blastema and reduced expression of Osr1 and SIX2, which mark nephron progenitor cells. Mutant mice are also characterized by defects in intermediate mesoderm-derived genital tissues with fewer mesonephric tubules and testicular Leydig cells, epithelial vacuolization in the postnatal corpus epididymis, and decreased serum testosterone levels and reduced fertility. Analysis of ALK3-dependent signaling effectors revealed lineage-specific reduction of phospho-p38 MAPK in metanephric mesenchyme and phospho-SMAD1/5/8 in the testis. Together, these results demonstrate a requirement for Alk3 in distinct progenitor cell populations derived from the intermediate mesoderm.


3-D augmented reality for MRI-guided surgery using integral videography autostereoscopic image overlay.
A 3-D augmented reality navigation system using autostereoscopic images was developed for MRI-guided surgery. The 3-D images are created by employing an animated autostereoscopic image, integral videography (IV), which provides geometrically accurate 3-D spatial images and reproduces motion parallax without using any supplementary eyeglasses or tracking devices. The spatially projected 3-D images are superimposed onto the surgical area and viewed via a half-silvered mirror. A fast and accurate spatial image registration method was developed for intraoperative i.v. image-guided therapy. Preliminary experiments showed that the total system error in patient-to-image registration was 0.90 +/- 0.21 mm, and the procedure time for guiding a needle toward a target was shortened by 75%. An animal experiment was also conducted to evaluate the performance of the system. The feasibility studies showed that augmented reality of the image overlay system could increase the surgical instrument placement accuracy and reduce the procedure time as a result of intuitive 3-D viewing.


Physiologic rate of carrier-mediated Ca2+ entry matches active extrusion in human erythrocytes.
The intracellular Ca2+ concentration of nearly all cells is kept at submicromolar levels. The magnitudes of transmembrane Ca2+ movement that maintain this steady state in the human red blood cell have long been debated. Although there is agreement that the physiologic extrusion of Ca2+ by the well-characterized Ca2+. ATPase amounts to 45 mumol/liter cells per h (1982. Nature (Lond.). 298:478-481), the reported passive entry rates in physiological saline (2-20 mumol/liter cells per h) are all substantially lower. This discrepancy could be due to incomplete inhibition of the pump in the previous measurements of Ca2+ entry. We therefore examined both rate and mechanism of entry after completely inactivating the pump. This required pretreatment with iodoacetamide (to lower the intracellular ATP concentration) and vanadate (to inhibit any residual Ca2+ pump activity). The rate of Ca2+ entry (53 mumol/liter cells per h) was now found to be comparable to the accepted extrusion rate. Entry closely obeyed Michaelis-Menten kinetics (Vmax = 321 +/- 17 nmol Ca/g dry wt per h, Km = 1.26 +/- 0.13 mM), was competitively inhibited by external Sr2+ (Ki = 10.8 +/- 1.2 mM), and was accelerated by intracellular Ca2+. 45Ca2+ efflux from these pump-inactivated cells was also accelerated by either external Ca2+ or Sr2+. These accelerating effects of divalent cations on the opposite (trans) face of the membrane rule out a simple channel. Substrate-gated channels are also ruled out: cells equilibrated with 45Ca2+ lost the isotope when unlabeled Ca2+ or Sr2+ was added externally. Thus, passive Ca2+ movements occur predominantly by a reversible carrier-mediated mechanism for which Sr2+ is an alternate substrate. The carrier's intrinsic affinity constants for Ca2+ and Sr2+, 1.46 and 0.37 mM-1, respectively, indicate that Ca2+ is the preferred substrate.


Increased Epithelial Oxygenation Links Colitis to an Expansion of Tumorigenic Bacteria.
Intestinal inflammation is a risk factor for colorectal cancer formation, but the underlying mechanisms remain unknown. Here, we investigated whether colitis alters the colonic microbiota to enhance its cancer-inducing activity. Colitis increased epithelial oxygenation in the colon of mice and drove an expansion of Escherichia coli within the gut-associated microbial community through aerobic respiration. An aerobic expansion of colibactin-producing E. coli was required for the cancer-inducing activity of this pathobiont in a mouse model of colitis-associated colorectal cancer formation. We conclude that increased epithelial oxygenation in the colon is associated with an expansion of a prooncogenic driver species, thereby increasing the cancer-inducing activity of the microbiota.IMPORTANCE One of the environmental factors important for colorectal cancer formation is the gut microbiota, but the habitat filters that control its cancer-inducing activity remain unknown. Here, we show that chemically induced colitis elevates epithelial oxygenation in the colon, thereby driving an expansion of colibactin-producing Escherichia coli, a prooncogenic driver species. These data suggest that elevated epithelial oxygenation is a potential risk factor for colorectal cancer formation because the consequent changes in the gut habitat escalate the cancer-inducing activity of the microbiota.


Recombinant human retinoic acid receptor alpha. Binding of DNA and synthetic retinoids to the protein expressed in Escherichia coli.
The human retinoic acid receptor alpha was expressed in Escherichia coli. The recombinant protein was found to be very unstable in several E. coli strains, probably due to proteolysis. Conditions were established to obtain reasonable amounts of active protein for ligand and DNA binding studies. The recombinant receptor showed the expected DNA binding activities in gel-retardation assays. Ligand binding properties were measured in a charcoal absorption assay. The dissociation constant for highly specific bound retinoic acid was found to be 0.67 nM. The affinity of several synthetic retinoids to the recombinant protein was determined and compared to their biological activity. Some of the values presented here differ significantly from those published earlier for the receptor or its isolated hormone-binding domain.


Incorporation of epitope-tagged viral sigma3 proteins to reovirus virions.
Tagging of viral capsid proteins is a powerful tool to study viral assembly; it also raises the possibility of using viral particles to present exogenous epitopes in vaccination or gene therapy strategies. The ability of reoviruses to induce strong mucosal immune response and their large host range and low pathogenicity in humans are some of the advantages of using reoviruses in such applications. In the present study, the feasibility of introducing foreign epitopes, "tags", to the sigma3 protein, a major component of the reovirus outer capsid, was investigated. Among eight different positions, the amino-terminal end of the protein appeared as the best location to insert exogenous sequences. Additional amino acids at this position do not preclude interaction with the micro1 protein, the other major constituent of the viral outer capsid, but strongly interfere with micro1 to micro1C cleavage. Nevertheless, the tagged sigma3 protein was still incorporated to virions upon recoating of infectious subviral particles to which authentic sigma3 protein was removed by proteolysis, indicating that micro1 cleavage is not a prerequisite for outer capsid assembly. The recently published structure of the sigma3- micro1 complex suggests that the amino-terminally inserted epitope could be exposed at the outer surface of viral particles.


[Autoreactivity of splenic cells of old rats and animals treated with hydrocortisone].
Spleen cells from 14-month-old mice are capable to induce GVH reaction in syngeneic recipients. Spleen cells obtained on the third day after hydrocortisone injection also induce GVH reaction in syngeneic recipients. The results suggest that T cells which suppress the activity of autoreactive lymphocytes are eliminated with age and after exposure to hydrocortisone.


The diurnal cycle of the smoky marine boundary layer observed during August in the remote southeast Atlantic.
Ascension Island (8° S, 14.5° W) is located at the northwestern edge of the south Atlantic stratocumulus deck, with most clouds in August characterized by surface observers as "stratocumulus and cumulus with bases at different levels", and secondarily as "cumulus of limited vertical extent" and occurring within a typically decoupled boundary layer. Field measurements have previously shown that the highest amounts of sunlight-absorbing smoke occur annually within the marine boundary layer during August. On more smoke-free days, the diurnal cycle in cloudiness includes a nighttime maximum in cloud liquid water path and rain, an afternoon cloud minimum, and a secondary late-afternoon increase in cumulus and rain. The afternoon low-cloud minimum is more pronounced on days with a smokier boundary layer. The cloud liquid water paths are also reduced throughout most of the diurnal cycle when more smoke is present, with the difference from cleaner conditions most pronounced at night. Precipitation is infrequent. An exception is the mid-morning, when the boundary layer deepens and liquid water paths increase. The data support a view that a radiatively enhanced decoupling persisting throughout the night is key to understanding the changes in the cloud diurnal cycle when the boundary layer is smokier. Under these conditions, the nighttime stratiform cloud layer does not always recouple to the sub-cloud layer, and the decoupling maintains more moisture within the sub-cloud layer. After the sun rises, enhanced shortwave absorption in a smokier boundary layer can drive a vertical ascent that momentarily couples the sub-cloud layer to the cloud layer, deepening the boundary layer and ventilating moisture throughout, a process that may also be aided by a shift to smaller droplets. After noon, shortwave absorption within smokier boundary layers again reduces the upper-level stratiform cloud and the sub-cloud relative humidity, discouraging further cumulus development and again strengthening a decoupling that lasts longer into the night. The novel diurnal mechanism provides a new challenge for cloud models to emulate. The lower free troposphere above cloud is more likely to be cooler, when boundary layer smoke is present, and lower free-tropospheric winds are stronger and more northeasterly, with both (meteorological) influences supporting further smoke entrainment into the boundary layer from above.


Activated, not resting, platelets increase leukocyte rolling in murine skin utilizing a distinct set of adhesion molecules.
Selectin-mediated tethering and rolling initiates the multi-step process of leukocyte extravasation which is crucial for the formation of an inflammatory infiltrate. We studied the impact of platelets on this process in the skin. Using intravital microscopy, we analyzed platelet interactions with cutaneous post-capillary venules of mouse ears and observed an increase in platelet rolling if platelets were activated (41.6+/-20.2% vs. 13.1+/-8.5% rolling of resting platelets). Experiments with P-selectin deficient mice and antibodies blocking either P-selectin, GPIIb/IIIa or GPIb showed that rolling depends on platelet PSGL-1 and GPIIb/IIIa on one hand, and endothelial P-selectin on the other. Next, formation of platelet-leukocyte aggregates was demonstrated by simultaneous observation of platelets and leukocytes in vivo utilizing a newly developed two-color technique. Aggregates increased overall leukocyte rolling (leukocytes alone: 27.4+/-11.2%, leukocytes with resting platelets: 25.3+/-10.2%, leukocytes with activated platelets 38.1+/-11.8%). To investigate if activated platelets may contribute to the pathogenesis of chronic cutaneous inflammation, platelet P-selectin expression was studied in 8 patients with psoriasis. A correlation between platelet P-selectin expression and disease severity was established. In summary, we show that activated, not resting, platelets increase leukocyte rolling in murine skin. This increased rolling is due to the aggregate formation of platelets with leukocytes. We also provide evidence for a potential role of this mechanism in the pathogenesis of chronic inflammatory skin diseases.