ONECUT2 is a driver of neuroendocrine prostate cancer.
Neuroendocrine prostate cancer (NEPC), a lethal form of the disease, is characterized by loss of androgen receptor (AR) signaling during neuroendocrine transdifferentiation, which results in resistance to AR-targeted therapy. Clinically, genomically and epigenetically, NEPC resembles other types of poorly differentiated neuroendocrine tumors (NETs). Through pan-NET analyses, we identified ONECUT2 as a candidate master transcriptional regulator of poorly differentiated NETs. ONECUT2 ectopic expression in prostate adenocarcinoma synergizes with hypoxia to suppress androgen signaling and induce neuroendocrine plasticity. ONEUCT2 drives tumor aggressiveness in NEPC, partially through regulating hypoxia signaling and tumor hypoxia. Specifically, ONECUT2 activates SMAD3, which regulates hypoxia signaling through modulating HIF1α chromatin-binding, leading NEPC to exhibit higher degrees of hypoxia compared to prostate adenocarcinomas. Treatment with hypoxia-activated prodrug TH-302 potently reduces NEPC tumor growth. Collectively, these results highlight the synergy between ONECUT2 and hypoxia in driving NEPC, and emphasize the potential of hypoxia-directed therapy for NEPC patients.


Urinary 1-hydroxypyrene concentration from Mexican children living in the southeastern region in Mexico.
Biomass combustion indoors has been associated with generation of various pollutants, such as carbon monoxide, formaldehyde, respirable particles, toluene, benzene and polycyclic aromatic hydrocarbons (PAHs), among others. In this context, the aim of this study was to evaluate the urinary 1-hydroxypyrene (as a biomarker exposure to PAHs) levels in children living in three states in Mexico. We evaluated children living in communities that use biomass fuels to cook and to heat homes in the next states: Chiapas, Oaxaca and Quintana Roo. We found similar levels of 1-OHP in urine of children living in the three studied states, with mean levels of 3.5 ± 1.0; 4.5 ± 2.8 and 4.4 ± 2.5 μg/L (geometric mean ± standard deviation), respectively for Chiapas, Oaxaca and Quintana Roo. In conclusion, our data indicate high exposure levels to PAHs in children living in the states studied in this work.


Surface oxidation of tin chalcogenide nanocrystals revealed by 119Sn-Mössbauer spectroscopy.
Narrow band gap tin(II) chalcogenide (SnS, SnSe, SnTe) nanocrystals are of high interest for optoelectronic applications such as thin film solar cells or photodetectors. However, charge transfer and charge transport processes strongly depend on nanocrystals' surface quality. Using (119)Sn-Mössbauer spectroscopy, which is the most sensitive tool for probing the Sn oxidation state, we show that SnS nanocrystals exhibit a Sn((IV))/Sn((II)) ratio of around 20:80 before and 40:60 after five minutes exposure to air. Regardless of the tin or sulfur precursors used, similar results are obtained using six different synthesis protocols. The Sn((IV)) content before air exposure arises from surface related SnS(2) and Sn(2)S(3) species as well as from surface Sn atoms bound to oleic acid ligands. The increase of the Sn((IV)) content upon air exposure results from surface oxidation. Full oxidation of the SnS nanocrystals without size change is achieved by annealing at 500 °C in air. With the goal to prevent surface oxidation, SnS nanocrystals are capped with a cadmium-phosphonate complex. A broad photoluminescence signal centered at 600 nm indicates successful capping, which however does not reduce the air sensitivity. Finally we demonstrate that SnSe nanocrystals exhibit a very similar behavior with a Sn((IV))/Sn((II)) ratio of 43:57 after air exposure. In the case of SnTe nanocrystals, the ratio of 55:45 is evidence of a more pronounced tendency for oxidation. These results demonstrate that prior to their use in optoelectronics further surface engineering of tin chalcogenide nanocrystals is required, which otherwise have to be stored and processed under inert atmosphere.


A Multilevel Conceptual Framework to Understand the Role of Food Insecurity on Antiretroviral Therapy Adherence in Low-Resource Settings: From Theory to Practice.
The objective of this study was to describe a multilevel conceptual framework to understand the role of food insecurity on antiretroviral therapy adherence. The authors illustrated an example of how they used the multilevel framework to develop an intervention for poor people living with HIV in a rural and low-resource community. The framework incorporates intrapersonal, interpersonal, and structural-level theories of understanding and changing health behaviors. The framework recognizes the role of personal, social, and environmental factors on cognition and behavior, with particular attention to ways in which treatment adherence is enabled or prevented by structural conditions, such as food insecurity.


Novel ACE2-IgG1 fusions with improved in vitro and in vivo activity against SARS-CoV2.
SARS-CoV2, the etiologic agent of COVID-19, uses ACE2 as a cell entry receptor. Soluble ACE2 has been shown to have neutralizing antiviral activity but has a short half-life and no active transport mechanism from the circulation into the alveolar spaces of the lung. To overcome this, we constructed an ACE2-human IgG1 fusion protein with mutations in the catalytic domain of ACE2. This fusion protein contained a LALA mutation that abrogates Fcrγ binding, but retains FcRN binding to prolong the half-life, as well as achieve therapeutic concentrations in the lung lavage. Interestingly, a mutation in the catalytic domain of ACE2, MDR504, completely abrogated catalytic activity, but significantly increased binding to SARS-CoV2 spike protein in vitro. This feature correlated with more potent viral neutralization in a plaque assay. Parental administration of the protein showed stable serum concentrations with a serum half-life of ~ 145 hours with excellent bioavailability in the epithelial lining fluid of the lung. Prophylactic administration of MDR504 significantly attenuated SARS-CoV2 infection in a murine model. These data support that the MDR504 hACE2-Fc is an excellent candidate for pre or post-exposure prophylaxis or treatment of COVID-19.


Coverage path planning for spraying drones.
The pandemic by COVID-19 is causing a devastating effect on the health of the global population. Currently, there are several efforts to prevent the spread of the virus. Among those efforts, cleaning and disinfecting public areas have become important tasks and they should be automated in future smart cities. To contribute in this direction, this paper proposes a coverage path planning method for a spraying drone, an unmanned aerial vehicle that has mounted a sprayer/sprinkler system, that can disinfect areas. State-of-the-art planners consider a camera instead of a sprinkler, in consequence, the expected coverage will differ in running time because the liquid dispersion is different from a camera's projection model. In addition, current planners assume that the vehicles can fly outside the target region; this assumption can not be satisfied in our problem, because disinfections are performed at low altitudes. Our method presents i) a new sprayer/sprinkler model that fits a more realistic coverage volume to the drop dispersion and ii) a planning method that efficiently restricts the flight to the region of interest avoiding potential collisions in bounded scenes. The algorithm has been tested in several simulation scenes, showing that it is effective and covers more areas with respect to two approaches in the literature. Note that the proposal is not limited to disinfection applications, but can be applied to other ones, such as painting or precision agriculture.


A Phase I clinical trial of chimeric antigen receptor-modified T cells in patients with relapsed and refractory lymphoma.
Aim: CD19 chimeric antigen receptor (CAR) T cells have been approved by the US FDA for treatment of relapsed and refractory (R/R) B-cell malignancies. Patients & methods: This study investigated the safety and efficacy of autologous 4-1BB costimulatory domain-engineered CD19 CAR-T cells in R/R B-cell lymphoma. Results: After CD19 CAR-T-cell infusion, severe cytokine release syndrome occurred in 28.6% (4/14) of the patients. The overall response rate was 77% with complete remission observed in 6/14 patients at 3 months. A higher tumor burden and grade 3-4 of myelosuppression after chemotherapy were associated with severe cytokine-release syndrome. Notably, combining CD19 CAR-T cells and PD-1 blockade, but not CD19 CAR-T cells alone, reduced intracranial tumor burden in a patient with central invasion of lymphoma. Conclusion: CD19 CAR-T cells could effectively induce tumor remission and PD-1 blockade might improve the efficacy in Chinese patients with R/R B-cell lymphoma.


Non-coordinate regulation of enzymes involved in transfer RNA metabolism in Escherichia coli.
During different steady state growth conditions in Escherichia coli the level of the three tRNA-modifying enzymes, the tRNA(m5Urd)-, tRNA(m1Guo)- and tRNA(mam5s2Urd)methyltransferase and of five aminoacyl-tRNA synthetases, the leucyl-, valyl-, isoleucyl-, arginyl- and threonyl-tRNA-synthetase, has been determined. It is shown that those two classes of tRNA affecting enzymes are not coordinately regulated and that even within these two groups of enzymes the constituents are regulated independently of each other. Furthermore it is demonstrated that none of the aminoacyl-tRNA synthetases and only one of the three tRNA-methyltransferases, the tRNA(m5Urd)methyltransferase, is under control of the relA+-gene.


FOXG1 expression shows correlation with neuronal differentiation in cerebellar development, aggressive phenotype in medulloblastomas, and survival in a xenograft model of medulloblastoma.
FOXG1 is a transcription factor that interacts with multiple signaling pathways and modulates neuronal differentiation in the telencephalon. Dysregulation of FOXG1 expression has been previously reported in medulloblastoma. In this study, we demonstrate a regional specific expression of FOXG1 and its colocalization with Nestin expression in the premigratory mitotically active (outer) layer of the external granular layer of the cerebellum. An inverse expression of the granular precursor cell markers, Math1 and Musashi1, in the inner nonmitotic migratory layer of the external granular layer and in the internal granular layer was observed. Furthermore, modulation of FOXG1 in the medulloblastoma cell line, DAOY, was associated with the induction of neuronal differentiation markers and significant changes in multiple signaling pathways regulating cell proliferation, differentiation, survival, and apoptosis. Additionally, we observed enhanced survival in intracerebellar mice xenografts injected with DAOY cells bearing shFOXG1 constructs versus shLuciferase construct. Overall, these findings suggest that down-modulation of FOXG1 is a prerequisite for the onset of neuronal differentiation during cerebellar development and that a decrease of FOXG1 in medulloblastoma cells offers a survival advantage in mice. We propose that the disruption of signaling pathways that promote mature neuronal differentiation by overexpressed FOXG1 is a contributing event in the neoplastic transformation of cerebellar stem cells.


Electroacupuncture Reduces Body Weight by Regulating Fat Browning-Related Proteins of Adipose Tissue in HFD-Induced Obese Mice.
Objective: This study investigated the influence of electroacupuncture (EA) and its potential underlying mechanisms on adipose tissue in obese mice. Methods: Three-week-old male C56BL/6 mice were randomly divided to feed or not to feed high-fat diet (HFD), named HFD group and chow diet (CD) group, respectively. After 12 weeks, CD and HFD mice were randomly divided into two groups, respectively, to receive or not receive EA for 4 weeks. Body weight (BW) was monitored. Intraperitoneal glucose tolerance test and metabolic chamber recordings were performed. Blood samples and adipose tissue were collected for the analysis of leptin, triglyceride levels, and fat browning-related proteins. Results: EA significantly reduced food intake, BW, and white adipose tissue (WAT)/BW ratio; decreased the adipocyte size and serum concentrations of triglyceride (TG) and cholesterol; and increased oxygen consumption in HFD mice. Compared with the CD mice, the HFD mice had elevated fasting serum glucose level and impaired glucose tolerance; however, these parameters were decreased by EA treatment. Meanwhile, EA promoted the protein and mRNA expressions of UCP1, PRDM16, and PGC-1α in adipose tissue, and activated sympathetic nerves via p-TH, A2AR, and β3AR in white adipose tissue. Conclusions: EA reduced food intake, BW, TG, and cholesterol, and improved glucose tolerance in HFD mice. This ameliorative effect of EA on obesity-related symptoms associated with its promoted adipose tissue plasticity via activating sympathetic nerves.


Interleukin-1 secretion by human alveolar macrophages stimulated with endotoxin is augmented by recombinant immune (gamma) interferon.
We investigated the role of the macrophage-activating factor gamma interferon (IFN) in the in vitro secretion of interleukin-1 (IL-1) from human alveolar macrophages. Macrophages were obtained from 6 normal volunteers and 5 patients by bronchoalveolar lavage. Macrophages were adhered to a plastic surface and then stimulated to secrete IL-1 with concentrations of endotoxin (LPS) ranging from 1 ng/ml to 10 micrograms/ml. Unstimulated macrophages served as a control. Recombinant gamma IFN was added to LPS-stimulated or unstimulated macrophage cultures in concentrations ranging from 0.25 to 1,000 U/ml. After 24 h of culture, supernatants were removed and IL-1 activity was measured by the thymocyte proliferation assay. Gamma IFN directly induced IL-1 secretion from macrophages of 2 of the subjects. In the presence of LPS, gamma IFN markedly augmented IL-1 secretion from macrophages of 10 of the 11 subjects. Gamma IFN could prime macrophages for increased response to LPS after 1 h of incubation. We conclude that gamma IFN provides a modulating signal for in vitro LPS-induced IL-1 secretion from alveolar macrophages and may enhance macrophage reactivity to inflammatory mediators in vivo.


Malnutrition accelerates the occurrence of infectious complications in patients with chronic kidney disease: A cross-sectional survey of 682 patients with chronic kidney disease.
To investigate the influencing factors of infectious complications in patients with chronic kidney disease (CKD) and provide a basis for clinical diagnosis and prognosis evaluation of CKD. A total of 682 patients with CKD were selected and divided into CKD stage 1-5 subgroups according to their glomerular filtration rate. Infectious complications, length of hospital stay, and total cost of hospitalization were recorded. The Global Leadership Initiative on Malnutrition (GLIM) diagnostic tool was used to assess the detection rate of malnutrition among patients. Univariate and multivariate analyses were performed in patients with and without infectious complications. The incidence rates of infectious complications in CKD stages 1-5 were 45.6%, 22.7%, 28.3%, 30.8%, and 40.4%, respectively. The overall detection rate of malnutrition among patients based on the GLIM criteria was 16.7%. The total detection rate of severe malnutrition was 14.2%, with all patients with severe malnutrition in CKD stages 3-5. The incidences of infectious complications in patients with and without malnutrition were 62.3% and 29%, respectively. Binary multivariate logistic regression analysis shows that malnutrition is a risk factor for infectious complications in patients with CKD, who are at 2.41 times higher risk than patients without malnutrition. There were significant differences in length of hospital stay and hospitalization costs between the patients with CKD with and without infectious complications (P < 0.01). Infectious complications are relatively common in patients with CKD. As CKD advances, the incidence of infectious complications increases. Moreover, malnutrition accelerates the occurrence of infectious complications in patients with CKD.


Unbiased gene expression analysis of the delayed fracture healing observed in Zucker diabetic fatty rats.
Impaired fracture repair in patients with type 2 diabetes mellitus (T2DM) is not fully understood. In this study, we aimed to characterize the local changes in gene expression (GE) associated with diabetic fracture. We used an unbiased approach to compare GE in the fracture callus of Zucker diabetic fatty (ZDF) rats relative to wild-type (WT) littermates at three weeks following femoral osteotomy. Zucker rats, WT and homozygous for leptin receptor mutation (ZDF), were fed a moderately high-fat diet to induce T2DM only in the ZDF animals. At ten weeks of age, open femoral fractures were simulated using a unilateral osteotomy stabilized with an external fixator. At three weeks post-surgery, the fractured femur from each animal was retrieved for analysis. Callus formation and the extent of healing were assessed by radiograph and histology. Bone tissue was processed for total RNA extraction and messenger RNA (mRNA) sequencing (mRNA-Seq). Radiographs and histology demonstrated impaired fracture healing in ZDF rats with incomplete bony bridge formation and an influx of intramedullary inflammatory tissue. In comparison, near-complete bridging between cortices was observed in Sham WT animals. Of 13,160 genes, mRNA-Seq analysis identified 13 that were differentially expressed in ZDF rat callus, using a false discovery rate (FDR) threshold of 10%. Seven genes were upregulated with high confidence (FDR = 0.05) in ZDF fracture callus, most with known roles in inflammation. These findings suggest that elevated or prolonged inflammation contributes to delayed fracture healing in T2DM. The identified genes may be used as biomarkers to monitor and treat delayed fracture healing in diabetic patients.


Elucidating the role of many-body forces in liquid water. I. Simulations of water clusters on the VRT(ASP-W) potential surfaces.
We test two new potentials for water, fit to vibration-rotation tunneling (VRT) data by employing diffusion quantum Monte Carlo simulations to calculate the vibrational ground-state properties of water clusters. These potentials, VRT(ASP-W)II and VRT(ASP-W)III, are fits of the highly detailed ASP-W (anisotropic site potential with Woermer dispersion) ab initio potential to (D(2)O)(2) microwave and far-infrared data, and along with the SAPT5s (five-site symmetry adapted perturbation theory) potentials, are the most accurate water dimer potential surfaces in the literature. The results from VRT(ASP-W)II and III are compared to those from the original ASP-W potential, the SAPT5s family of potentials, and several bulk water potentials. Only VRT(ASP-W)III and the spectroscopically "tuned" SAPT5st (with N-body induction included) accurately reproduce the vibrational ground-state structures of water clusters up to the hexamer. Finally, the importance of many-body induction and three-body dispersion are examined, and it is shown that the latter can have significant effects on water cluster properties despite its small magnitude.


Iron oxides alter methanogenic pathways of acetate in production water of high-temperature petroleum reservoir.
Acetate is a key intermediate in anaerobic crude oil biodegradation and also a precursor for methanogenesis in petroleum reservoirs. The impact of iron oxides, viz. β-FeOOH (akaganéite) and magnetite (Fe3O4), on the methanogenic acetate metabolism in production water of a high-temperature petroleum reservoir was investigated. Methane production was observed in all the treatments amended with acetate. In the microcosms amended with acetate solely about 30% of the acetate utilized was converted to methane, whereas methane production was stimulated in the presence of magnetite (Fe3O4) resulting in a 48.34% conversion to methane. Methane production in acetate-amended, β-FeOOH (akaganéite)-supplemented microcosms was much faster and acetate consumption was greatly improved compared to the other conditions in which the stoichiometric expected amounts of methane were not produced. Microbial community analysis showed that Thermacetogenium spp. (known syntrophic acetate oxidizers) and hydrogenotrophic methanogens closely related to Methanothermobacter spp. were enriched in acetate and acetate/magnetite (Fe3O4) microcosms suggesting that methanogenic acetate metabolism was through hydrogenotrophic methanogenesis fueled by syntrophic acetate oxidizers. The acetate/β-FeOOH (akaganéite) microcosms, however, differed by the dominance of archaea closely related to the acetoclastic Methanosaeta thermophila. These observations suggest that supplementation of β-FeOOH (akaganéite) accelerated the production of methane further, driven the alteration of the methanogenic community, and changed the pathway of acetate methanogenesis from hydrogenotrophic methanogenesis fueled by syntrophic acetate oxidizers to acetoclastic.


Horizontal root fracture in a maxillary central incisor: a case report.
Horizontal root fractures usually are characterized by a fracture line that is perpendicular to the long axis of the tooth's root. Root fractures are diagnosed through clinical and radiographic examination. Treatment depends on the position of the fracture and the extent of root involvement. Successful treatment depends on correct diagnosis, clinical management, and radiographic follow-up. This article describes the successful treatment (with a 3-year follow-up period) of a permanent maxillary central incisor with horizontal root fracture treated with calcium hydroxide, in which the coronal segment was necrotic and the apical segment remained vital.


[The army medical service: past, present, and future].
The changes in the geopolitical situation after the dissolution of the Warsaw-pact forced many western governments to reform drastically their armed forces. Moreover, the postponing of the conscription gave this restructuring a very particular dimension. Despite all this alterations the fundamental missions of the Medical Service of the Armed Forces have not changed. They still consist of maintaining the operational readiness of the troops, taking care of the wounded and reducing physical and mental handicaps. Therefore an entity of levels of consistent and capable medical care is needed. The rational construction of this complete and fulfilling chain of help, from point of wounding to the discharge from the hospital on national territory is the right to exist for an autonomous Medical Service. It goes without saying that this treatment must have the same quality standards as those used in the civilian health care system. Taking this into account the Medical Service of the Armed Forces integrated its means into civilian services for urgency and disaster medicine. In the scientific field there are a lot of close contacts and agreements with several universities in order to deal with the military aspects of the medical research. It is in fact the ethical duty of the Medical Service to provide for optimal care to the Armed Forces. Can a nation omit this? Will arguments of budgetary kind and the relative international "detente" again do forget us the lessons learned from the past? Is an autonomous Medical Service a superfluous luxury?


Distraction of the growth plate. Experiments in pigs and sheep.
Distraction of the growth plate of the distal radius was performed on 15 pigs and 10 sheep of growing age. The separation of the plate from the metaphysis occurred within 4-6 days after the operation. The distraction rate was 1-1.5 mm/day and was continued for 1-3 weeks. Consolidation of the separation area occurred within 6 weeks. Oxytetracycline bone labelling showed that the ossification developed toward the centre of the separation line from the metaphysis and from the inner layer of the periosteum as well as from the physeal side. The growth of the bone continued after the distraction procedure and in most cases the growth plate was open when examined radiologically after the distraction.


Dilemma of diagnosing thoracic sarcoidosis in tuberculosis endemic regions: An imaging-based approach. Part 1.
Sarcoidosis is a multi-systemic disorder of unknown etiology, although commonly believed to be immune-mediated. Histologically, it is characterized by noncaseating granuloma which contrasts against the caseating granuloma seen in tuberculosis (TB), an infectious disease that closely mimics sarcoidosis, both clinically as well as radiologically. In TB-endemic regions, the overlapping clinico-radiological manifestations create significant diagnostic dilemma, especially since the management options are markedly different in the two entities. Part 1 of this review aims to summarize the clinical, laboratory, and imaging features of sarcoidosis, encompassing both typical and atypical manifestations, in an attempt to distinguish between the two disease entities.


Massive bone marrow involvement in an end stage renal failure case with erythropoietin-resistant anemia and primary hyperoxaluria.
Primary hyperoxaluria is a rare autosomal recessive disorder. Type 1 PH is the most common form and develops due to a defect in a liver specific enzyme the alanine aminotransferase enzyme. As a result of the enzyme deficiency, there is an overproduction of oxalate and excessive urinary excretion. Recurrent urolithiasis and nephrocalcinosis are the most important findings of the disorder and often at the beginning end-stage renal disease develops. This report presents a case backed up by literature of a patient with end stage renal failure and erythropoietin-resistant anaemia whose bone marrow biopsy showed crystal deposition which received delayed diagnosis of oxalosis.


Lichen planus pemphigoides induced by anti-PD-1 antibody: A case only involved in oral mucosa with excellent topical treatment efficiency.
Lichen planus pemphigoides (LPP) is a rare autoimmune subepidermal disease that can occur in patients receiving immune checkpoint inhibitors. Its clinical manifestations are combined with the characteristics of lichen planus with bullous pemphigoid that can occur on either skin or oral mucosa. It should be noted that oral LPP is very rare. Here, we report a novel case of oral LPP induced by an anti-PD-1 agent. The patient presented with typical clinical features in oral mucosa, and the diagnosis was based on histopathology and immunological studies. Given that the patient was receiving an anti-PD-1 agent, topical therapy was chosen, and a nice therapeutic effect was obtained. No significant recurrence was observed after a 2-year follow-up. A good and stable therapeutic effect achieved by rapid and local symptomatic medication suggests that accurate and sensitive diagnosis is necessary.


Selective retention of recombinant plasmids coding for human insulin.
Plasmids may be lost from Escherichia coli K-12 hosts that are cultured without selection for plasmid retention. This is particularly true for chimeric plasmids that incorporate genes for human insulin into vectors derived from pBR322. The cIts857 gene of bacteriophage lambda was inserted into the bla gene of the human-insulin-coding plasmids, pIA7 delta 4 delta 1, pIB7 delta 4 delta 1 and pHI7 delta 4 delta 1, generating the new plasmids pPR17, pPR18 and pPR19, respectively, which produced the thermosensitive lambda repressor. The cI gene was downstream from the pM and pbla promoters, so that it may have been expressed from either or both promoters. Separate E. coli K-12 RV308 host strains containing the new recombinants were lysogenized with the repressor-defective bacteriophage lambda cI90. Loss of the plasmid from the lysogens causes concomitant loss of the lambda repressor and cell death, because the prophage is induced to enter the lytic growth cycle. The system effectively forces retention of the plasmid in all viable cells in the culture.


[Inhalation lesions in the burn patient].
Inhalation injuries are currently the factor most responsible for mortality in thermally injured patients. Inhalation injuries may occur independently, but generally occur together with skin burn. Smoke inhalation affects all levels of the respiratory system and the extent of the inhalation injury depends on the duration, exposure, amount and toxicity of the fume temperature, concentration and solubility of toxic gases, the occurrence of the accident in a closed space and pre-existing diseases. Smoke inhalation also induces changes in the systemic organs with the need for more fluid for resuscitation. Systemic vasoconstriction, with an elevation in systemic vascular resistance, a fall in myocardial contractility and a great increase in lymphatic flow in soft tissue are the most important changes in systemic organs. On admission of a burn patient there is a high suspicion of inhalation injury when there are signs and symptoms such as hoarseness, strides, dyspnea, carbonaceous sputum, anxiety or disorientation, with or without face burns. The patient with these findings has partial airway obstruction and there is substantial risk complete airway obstruction occurring of secondary to the edema. Patients with suspected inhalation injury should be intubated so as to maintain airway patency and avoid a total obstruction. This group of patients frequently develop respiratory failure with the need for mechanical ventilatory support. Nosocomial infections, sepsis and multiple organ system failure may occur. Late complications of inhalation injury are tracheitis, tracheal stenosis or tracheomalacia and chronic airway disease, which is relatively rare. Early diagnosis of inhalation injury and treatment in a Burn Unit by a group of highly motivated clinicians and a good team of nurses is essential in order to decrease the morbidity and mortality related to inhalation injury.


Isolation, characterization and application of a lytic phage vB_VspM_VS1 against Vibrio splendidus biofilm.
Vibrio splendidus is a common pathogen in the ocean that infects Apostichopus japonicus, Atlantic salmon and Crassostrea gigas, leading to a variety of diseases. In this study, a virulent phage vB_VspM_VS1, which infects V. splendidus, was isolated from aquaculture ponds in Dalian, China, and it belongs to the family Straboviridae in the order Caudoviricetes. vB_VspM_VS1 had an adsorption rate of 96% in 15 min, a latent period of 65 min, and a burst size of 140 ± 6 PFU/cell. The complete genome of phage vB_VspM_VS1 consists of a linear double-stranded DNA that is 248,270 bp in length with an average G + C content of 42.5% and 389 putative protein-coding genes; 116 genes have known functions. There are 4 tail fiber genes in the positive and negative strands of the phage vB_VspM_VS1 genome. The protein domain of the phage vB_VspM_VS1 tail fibers was obtained from the Protein Data Bank and the SMART (http://smart.embl.de) database. Bacterial challenge tests revealed that the growth of V. splendidus HS0 was apparently inhibited (OD600 < 0.01) in 12 h at an MOI of 10. In against biofilms, we also showed that the OD570 value of the vB_VspM_VS1-treated group (MOI = 1) decreased significantly to 0.04 ± 0.01 compared with that of the control group (0.48 ± 0.08) at 24 h. This study characterizes the genome of the phage vB_VspM_VS1 that infects the pathogenic bacterium V. splendidus of A. japonicus.


Effects of hormone deprivation and 2-methoxyestradiol combination therapy on hormone-dependent prostate cancer in vivo.
2-Methoxyestradiol (2-ME) has potent antiproliferative effects on cancer cells. Its utility alone or in combination with other therapies for treating prostate cancer, however, has not been fully explored. Androgen-dependent and independent human prostate cancer cells were examined in vivo for their response to combination therapy. Efficacy was assessed by terminal deoxynucleotide transferase-mediated dUTP nick-end labeling assay and measuring microvessel density (MVD) in excised tumors. Animals harboring hormone-dependent tumors treated with 2-ME alone, androgen deprivation therapy alone, or the combination of the two had a 3.1-fold, 5.3-fold, and 10.1-fold increase in apoptosis, respectively. For hormone-independent tumors, treatment with 2-ME resulted in a 2.43-fold increase in apoptosis and a 73% decrease in MVD. 2-ME was most effective against hormone-dependent tumors in vivo and combination therapy resulted in a significant increase in efficacy compared to no treatment controls and trended toward greater efficacy than either 2-ME or androgen deprivation alone. Combination therapy should be investigated further as an additional therapeutic option for early prostate cancer.


From self-assembly to noncovalent synthesis of programmable porphyrins' arrays in aqueous solution.
Self-organization is the driving force that led to the evolution of life. Rationalization of the spontaneous self-assembly paradigm will offer tremendous potentialities to obtain a wide variety of complex systems, having specific functionality and properties. Herein, we will propose an overview of the developments in non-covalent syntheses of multi-porphyrin supramolecular species in aqueous solution. This work took inspiration from the pioneering studies aimed at rationalizing the spontaneous aggregation processes, governed by conventional solution properties (i.e. pH, ionic strength, and concentration). The more recent chemical strategies, to hierarchically manipulate the cooperative nature of weak interactions to design and synthesize supramolecular entities having pre-determined structure and properties, demonstrate the feasibility to attain, in a reproducible manner, molecular organization to supramolecular levels. In particular, calixarene-porphyrin species represents concrete evidence of a quantitative complexation, governed by precise hierarchical rules, which together with a rational functionalization of the molecular components leads to supramolecular entities of well-defined and tunable stoichiometry. These systems, thus, represent fertile ground to envisage and implement controlled self-organization strategies as bottom up methodologies to obtain supramolecular nanostructures and smart nanomachines.


Carrier transport dynamics in Mn-doped CdSe quantum dot sensitized solar cells.
In this work quantum dot sensitized solar cells (QDSSCs) were fabricated with CdSe and Mn-doped CdSe quantum dots (QDs) using the SILAR method. QDSSCs based on Mn-doped CdSe QDs exhibited improved incident photon-to-electron conversion efficiency. Carrier transport dynamics in the QDSSCs were studied using the intensity modulated photocurrent/photovoltage spectroscopy technique, from which transport and recombination time constants could be derived. Compared to CdSe QDSSCs, Mn-CdSe QDSSCs exhibited shorter transport time constant, longer recombination time constant, longer diffusion length, and higher charge collection efficiency. These observations suggested that Mn doping in CdSe QDs could benefit the performance of solar cells based on such nanostructures.


Presence of lymphocytic infiltrate cytotoxic T lymphocyte CD3+, CD8+, and immunoscore as prognostic marker in patients after radical cystectomy.
Tumor-Infiltrating Lymphocytes (TILs) has been shown to be essential to predict disease outcome in several types of cancers. Moreover, the distribution of cytotoxic T lymphocytes (CD8+) and T cells in general (CD3+) have been used to establish an Immunoscore, as a new cancer prognosticator for survival in colon and lung cancer. In bladder cancer, immune activation has been shown to be associated with genomic subtypes of muscle invasive bladder cancer (MIBC). We sought to evaluate the prognostic impact of these immune cell types in MIBC patients treated with radical cystectomy. For this purpose, cystectomy sections (n = 67) with identifiable invasive margin were selected and stained for CD8+ and CD3+ tumour infiltrating lymphocytes (TILs); both tumor core (CT) and invasive margin (IM) were assessed. Immunoscore was calculated based on previously defined criteria and used to illustrate differences in survival. High density of CD8IM TILs was associated with better disease-free (DFS) (P = 0.01) and overall survival (OS) (P = 0.02) whereas CD3IM TILs were associated with better OS (P = 0.05). Immunoscore was associated with improved DFS (P = 0.02) and OS (P = 0.05). The expression of cytotoxic T cells (CD8+ T cells) in TCGA bladder cancer was also investigated from RNA-Seq profiles of 344 cases. T cell cytotoxicity associated genes (n = 113) were derived from MSig GSEA database. Luminal (n = 121) and basal (n = 68) samples were used to evaluate expression differences. Differential expression (P<0.05) of cytotoxic T cell genes was noted across different molecular subsets of bladder cancer within TCGA analysis. Our data suggests host immune system appears to play a valuable prognostic role in MIBC.


Solution conformation and dynamics of the HIV-1 integrase core domain.
The human immunodeficiency virus type 1 (HIV-1) integrase (IN) is a critical enzyme involved in infection. It catalyzes two reactions to integrate the viral cDNA into the host genome, 3' processing and strand transfer, but the dynamic behavior of the active site during catalysis of these two processes remains poorly characterized. NMR spectroscopy can reveal important structural details about enzyme mechanisms, but to date the IN catalytic core domain has proven resistant to such an analysis. Here, we present the first NMR studies of a soluble variant of the catalytic core domain. The NMR chemical shifts are found to corroborate structures observed in crystals, and confirm prior studies suggesting that the alpha4 helix extends toward the active site. We also observe a dramatic improvement in NMR spectra with increasing MgCl(2) concentration. This improvement suggests a structural transition not only near the active site residues but also throughout the entire molecule as IN binds Mg(2+). In particular, the stability of the core domain is linked to the conformation of its C-terminal helix, which has implications for relative domain orientation in the full-length enzyme. (15)N relaxation experiments further show that, although conformationally flexible, the catalytic loop of IN is not fully disordered in the absence of DNA. Indeed, automated chemical shift-based modeling of the active site loop reveals several stable clusters that show striking similarity to a recent crystal structure of prototype foamy virus IN bound to DNA.


Formation of granulation tissue in intra-abdominal viscose cellulose sponges. An experimental study in rats.
Formation of intra-abdominal granulation tissue was studied in 32 rats 2, 4, 7 or 21 days after implantation of viscose cellulose sponges. The concentrations of DNA, RNA-ribose, uronic acids, hexosamines, nitrogen and collagen were then measured in the sponges. The rats' body weight fell by 10-15% in the first week, but was regained within 3 weeks. The sponges induced dense intra-abdominal adhesions, reflected in highly active connective tissue formation. This process seemed analogous to that seen in granuloma formation in subcutaneous tissue or wound healing, but the synthesis of macromolecules was especially pronounced in the peritoneal cavity. The experimental model offers a new means for studying intra-abdominal adhesions and permits evaluation of factors claimed to have stimulating, inhibiting or no effect on intra-abdominal adhesion formation.


Mechanical properties of a self-assembling oligopeptide matrix.
We have begun studies of a novel type of biomaterial derived from a recently-discovered class of ionic self-complementary oligopeptides. These short peptides (typically 8, 16, 24, or 32 amino acid residues with internally-repeating sequences) self-assemble in aqueous salt solution into three-dimensional matrices capable of favorable interactions with cells, and offer promise for useful bioengineering design based on rational changes in sequence. In this paper we present preliminary results on mechanical properties, combining experimental and theoretical approaches, of one particular example of these peptide materials, EFK8. The static elastic modulus was measured using an apparatus designed to allow sample fabrication and mechanical testing in the same system with the sample in aqueous solution. The material microstructure was examined by SEM and the measurements interpreted with the aid of a model for cellular solids. Values for the elastic modulus increased from 1.59 +/- 0.06 to 14.7 +/- 1.0 kPa for peptide concentrations increasing from 2.7 to 10 mg ml-1. SEM photographs showed the microstructure to consist of a relatively homogeneous lattice with fiber thickness of 10-30 nm independent of peptide concentration, but with fiber density increasing with peptide concentration. This behavior is consistent with scaling predictions from the cellular solids model and yields an estimate for the individual fiber elastic modulus in the range of 1-20 MPa. We therefore have provided some initial physical principles for guiding improvement of the mechanical properties of these new materials.


[Effect of antipurine antibodies on the immunodepressive activity of 6-mercaptopurine].
In repeated injections of 6-mercaptopurine (6-MP) covalently bound with protein (6-MP-P) antibodies binding 6-MP appeared in rabbits. In such animals free 6-MP produced a lesser immunodepressive and toxic action. Injection to mice immunized with sheep erythrocytes (SE) of rabbit serum containing antibodies against 6-MP not only failed to lead to reduction of the immunodepressive action of this cytostatic, but, on the contrary, intensified the inhibition of antibody formation to the SE. Injection to mice of 6-MP bound with rabbit antiserum in vitro also enhanced its immunodepressive activity. Evidently own antibodies neutralized 6-MP, whereas foreign ones, serving as an antigen, apparently caused its accumulation in the macrophagal lymphocytic complex.


Accelerated Hantzsch electrospray synthesis with temporal control of reaction intermediates.
Complex chemical reactions can occur in electrosprayed droplets on the millisecond time scale. The Hantzsch synthesis of 1,4-dihydropyridines was studied in this way using on-line mass spectral analysis to optimize conditions and characterize the product mixture. Changing the distance between the nanospray source and the MS inlet allowed exploration of reaction progress as a function of droplet time-of-flight. Desolvation of the charged microdroplets is associated with transformation from starting material to intermediates and eventually to product as the distance is increased. Results of the on-line experiments require a termination step that discontinuously completes the desolvation process and allows the generated gaseous ions to be used to characterize the state of the system at a particular time. The intermediates seen correspond to those known to occur in the bulk solution-phase reaction. Off-line collection of the sprayed reaction mixture allowed the recovery of 250 mg h-1 of desired reaction product from a single sprayer, permitting characterization by NMR and other standard methods. A thin film version of the accelerated reaction is described and it could be controlled through the temperature of the collection surface.


Enhancing Efficiency of Nonfullerene Organic Solar Cells via Using Polyelectrolyte-Coated Plasmonic Gold Nanorods as Rear Interfacial Modifiers.
Sufficient sunlight absorption and exciton generation are critical for developing efficient nonfullerene organic solar cells (OSCs). In this work, polyelectrolyte polystyrenesulfonate (PSS)-coated plasmonic gold nanorods (GNRs@PSS) were incorporated, for the first time, into the inverted nonfullerene OSCs as rear interfacial modifiers to improve sunlight absorption and charge generation via the near-field plasmonic and backscattering effects. The plasmonic GNRs effectively improved the sunlight absorption and enhanced the charge generation. Meanwhile, the negatively charged PSS shell ensured the uniform dispersion of the GNRs on the surface of the photoactive layer, optimized the interfacial contact, and further promoted the hole transport to the electrode. These concerted synergistic effects augmented the efficiency (10.11%) by nearly 20% relative to the control device (8.47%). Remarkably, the ultrathin (∼2.2 nm) organic layer on the surface of GNRs was closely examined by acquiring the carbon contrast image through energy-filtered transmission electron microscopy (EF-TEM), which clearly confirmed the coating uniformity from the side to end-cap of GNRs. The surface plasmon resonance (SPR) effect of the GNRs@PSS on the surface of the photoactive layer was unprecedentedly mapped by photoinduced force microscopy (PiFM) under the illumination of a tunable wavelength supercontinuum laser mimicking sunlight. Furthermore, investigations into the effect of size, surface coverage, and incorporation location of GNRs@PSS on the performance of OSCs revealed that the appropriate design and incorporation of the plasmonic nanostructures are crucial, otherwise the performance can be decreased, as evidenced in the case of front interface integration.


Deep-ultraviolet-light-driven reversible doping of WS2 field-effect transistors.
Improvement of the electrical and photoelectric characteristics is essential to achieve an advanced performance of field-effect transistors and optoelectronic devices. Here we have developed a doping technique to drastically improve electrical and photoelectric characteristics of single-layered, bi-layered and multi-layered WS2 field-effect transistors (FET). After illuminating with deep ultraviolet (DUV) light in a nitrogen environment, WS2 FET shows an enhanced charge carrier density, mobility and photocurrent response. The threshold voltage of WS2 FET shifted toward the negative gate voltage, and the positions of E and A1g peaks in Raman spectra shifted toward lower wavenumbers, indicating the n-type doping effect of the WS2 FET. The doping effect is reversible. The pristine characteristics of WS2 FET can be restored by DUV light illumination in an oxygen environment. The DUV-driven doping technique in a gas environment provides a very stable, effective, easily applicable way to enhance the performance of WS2 FET.


Using Case Studies to Promote Student Engagement in Primary Literature Data Analysis and Evaluation.
Analyzing and evaluating primary literature data is a common learning objective in undergraduate neuroscience courses. However, students with more clinically focused career goals often dismiss the relevance of evaluating basic neuroscience literature. Here, we describe using case studies to promote student engagement in primary literature in a cellular and molecular neuroscience course. Two example literature-based case studies are provided: Untwisting Pretzel Syndrome, a neurodevelopment case exploring synapse formation in a pretzel syndrome patient, and The Trials of ALS, a neurodegeneration case exploring axon degeneration and repair in an amyotrophic lateral sclerosis patient. These cases were assigned after neurodevelopment and neurodegeneration lectures covering key concepts. Both cases begin by introducing the patient and hypothesizing symptoms and diagnoses, followed by scenes incorporating primary data to illustrate disease pathogenesis and treatments. Students complete questions embedded in these cases as homework, and class time is used to discuss their answers. Discussion emphasizes that there can be multiple "correct" answers, and the best answers are accurate and well-supported. Accordingly, students edit their answers in class, and these annotations are factored into a pass/fail grade on the case. Additional scenes and questions from the same case studies are used on the course's take-home exams, thereby allowing students to practice primary data analysis and evaluation before a graded assignment. Student evaluations support literature-based case studies as an effective learning tool, with students identifying cases as the most valuable aspect of the course, and reporting increased confidence in understanding cellular and molecular neuroscience.


Kinin signal transduction: role of phosphoinositides and eicosanoids.
Kinins elicit prostaglandin and inositol phosphate production in 3T3 fibroblasts through stimulation of B2 receptors. Prostaglandin synthesis is maximum by 5 min, whereas inositol phosphate production continues for longer than 30 min. Prostaglandin synthesis is stimulated by phospholipase A2, which releases arachidonate from phospholipids, whereas a phosphatidylinositol-specific phospholipase C catalyzes formation of equimolar amounts of inositol phosphate and diacylglycerol. Stimulation of these two second-messenger systems occurs through independent pathways: (a) dexamethasone inhibits prostaglandin formation by inhibiting phospholipase A2, and, to a lesser degree, cyclooxygenase, but is without effect on inositol phosphate production; (b) neomycin inhibits inositol phosphate production without affecting prostaglandin synthesis; (c) phorbol esters inhibit inositol phosphate production while augmenting prostaglandin synthesis; and (d) indomethacin inhibits prostaglandin synthesis but does not affect inositol phosphate production. At later times (greater than 10 min), the two pathways interact. Stimulation with one agonist to increase diacylglycerol results in augmentation of prostaglandin synthesis in response to a second agonist. Inositol phosphates cause release of calcium from intracellular stores. Prostaglandins stimulate (by binding to their own receptors) adenylate cyclase to increase cAMP. Additionally, prostaglandins increase intracellular free calcium by increasing influx of extracellular calcium. Both inositol phosphates and prostaglandins play roles in mitogenesis in these cells.


Artificial skin: past, present and future.
The integrity of the skin should be restored as soon as possible whenever the skin gets wounded. Recent research has revealed some of the complex pathways in wound healing. Based on this knowledge, researchers have been looking for better skin substitutes to treat difficultly healing or large wounds. Some of these highly sophisticated wound dressings, also known as bio-dressings, contain material of human or animal origin, e.g. cultured skin cells. Although the ideal skin substitute has not been established yet, the currently available bio-dressings help clinicians close difficultly healing skin wounds.


Sympathetic nerve reconstruction for compensatory hyperhidrosis after sympathetic surgery for primary hyperhidrosis.
We performed sympathetic nerve reconstruction using intercostal nerve in patients with severe compensatory hyperhidrosis after sympathetic surgery for primary hyperhidrosis, and analyzed the surgical results. From February 2004 to August 2007, sympathetic nerve reconstruction using intercostal nerve was performed in 19 patients. The subjected patients presented severe compensatory hyperhidrosis after thoracoscopic sympathetic surgery for primary hyperhidrosis. Reconstruction of sympathetic nerve was performed by thoracoscopic surgery except in 1 patient with severe pleural adhesion. The median interval between the initial sympathetic surgery and sympathetic nerve reconstruction was 47.2 (range: 3.5-110.7) months. Compensatory sweating after the reconstruction surgery improved in 9 patients, and 3 out of them had markedly improved symptoms. Sympathetic nerve reconstruction using intercostal nerve may be one of the useful surgical options for severe compensatory hyperhidrosis following sympathetic surgery for primary hyperhidrosis.


Ledge formation: review of a great challenge in endodontics.
Failure to grasp the rationale behind cleaning and shaping concepts can increase the occurrence of needless complications, such as ledges. A ledge is created when the working length can no longer be negotiated and the original pathway of the canal has been lost. Extension of the access cavity to provide unobstructed access to the root canals, precurving and not forcing instruments, using NiTi files, using passive step-back and balanced force techniques, and instrumenting the canal to its full length will all help to prevent ledge formation. Initial negotiation and bypassing the ledge can be achieved using a small file with a distinct curve at the tip, whereas a slight rotation motion of the file combined with a ''picking'' motion can often help advance the instrument. A review of the literature and a discussion of the options for prevention, recognition and managing the ledge are presented.


[Flexibility of working hours and health: towards ergonomics of working time].
The search for ways and methods able to increase the "flexibility" of working hours deal with several forms of intervention that depend on political choices and work management, according to specific interests and needs of the companies, the individual worker and the whole society. The main problem on the carpet is to evaluate whether that interferes with worker's health and well-being. According to the data of the last European Survey on Working Conditions (EURF 2000), it appears the workers engaged in working hours different from the traditional day work are nowadays the vast majority of the population; moreover, organisational forms which allow more flexibility, that is more autonomy, in working time arrangement are associate to better health and well-being.


Ultrastructural changes in Raillietina (Platyhelminthes: cestoda), exposed to sulfonoquinovosyldiacylglyceride (SQDG), isolated from Neem (Azadirachta indica).
Neem (Azadirachta indica), has been known to be a curative for various ailments and diseases in the traditional Indian medicinal system from times immemorial. A glycolipid sulfonoquinovosyldiacylglyceride (SQDG) isolated from the leaves of neem has been found to be a proactive antibacterial and antiviral agent in previous studies. The current communication pertains to the anthelmintic activity of SQDG in vitro against a model cestode Raillietina spp. The results of efficacy tests showed a paralysis time of 1.0 ± 0.1 and 0.7 ± 0.01 h, whereas death time of 1.6 ± 0.3 and 0.9 ± 0.02 h, following treatments with dosages of 0.5 and 1.0 mg/mL, respectively. The scanning electron microscopic studies showed significant and unique changes in the ultrastructure of the worms with prominent breakages and furrows on the surface.


Operations on the pancreas; some technical considerations.
Many diseases of the pancreas formerly universally considered fatal, now are known to be amenable to surgical therapy. Pancreatic cysts, pseudocysts, calculi, inflammation, and benign and malignant tumors all can be dealt with effectively by operation. In this presentation various surgical techniques which have proved to be of therapeutic importance are considered.


High-field EPR and ESEEM investigation of the nitrogen quadrupole interaction of nitroxide spin labels in disordered solids: toward differentiation between polarity and proticity matrix effects on protein function.
The combination of high-field electron paramagnetic resonance (EPR) with site-directed spin labeling (SDSL) techniques employing nitroxide radicals has turned out to be particularly powerful in revealing subtle changes of the polarity and proticity profiles in proteins enbedded in membranes. This information can be obtained by orientation-selective high-field EPR resolving principal components of the nitroxide Zeeman (g) and hyperfine ( A) tensors of the spin labels attached to specific molecular sites. In contrast to the g- and A-tensors, the (14)N ( I = 1) quadrupole interaction tensor of the nitroxide spin label has not been exploited in EPR for probing effects of the microenvironment of functional protein sites. In this work it is shown that the W-band (95 GHz) high-field electron spin echo envelope modulation (ESEEM) method is well suited for determining with high accuracy the (14)N quadrupole tensor principal components of a nitroxide spin label in disordered frozen solution. By W-band ESEEM the quadrupole components of a five-ring pyrroline-type nitroxide radical in glassy ortho-terphenyl and glycerol solutions have been determined. This radical is the headgroup of the MTS spin label widely used in SDSL protein studies. By DFT calulations and W-band ESEEM experiments it is demonstrated that the Q(yy) value is especially sensitive to the proticity and polarity of the nitroxide environment in H-bonding and nonbonding situations. The quadrupole tensor is shown to be rather insensitive to structural variations of the nitroxide label itself. When using Q(yy) as a testing probe of the environment, its ruggedness toward temperature changes represents an important advantage over the g xx and A(zz) parameters which are usually employed for probing matrix effects on the spin labeled molecular site. Thus, beyond measurenments of g xx and A(zz) of spin labeled protein sites in disordered solids, W-band high-field ESEEM studies of (14)N quadrupole interactions open a new avenue to reliably probe subtle environmental effects on the electronic structure. This is a significant step forward on the way to differentiate between effects from matrix polarity and hydrogen-bond formation.


Transcriptomal profiling of bovine ovarian granulosa and theca interna cells in primary culture in comparison with their in vivo counterparts.
In vitro culture of ovarian granulosa cells and theca cells has been very important for our understanding of their function and regulation. One of the most eagerly sought attributes of cell culture is the use of chemically-defined conditions. However, even under such in vitro conditions cell behaviour could differ from the in vivo situation because of differences in oxygen tension, nutrients, adhesion matrix and other factors. To examine this further we compared the transcriptomes of both granulosa cells and cells from the theca interna that were cultured in what are arguably the best in vitro conditions for maintaining the 'follicular' phenotypes of both tissue types, as displayed by their respective freshly-isolated counterparts. The array data analysed are from recently published data and use the same sizes of bovine follicles (small antral 3-6 mm) and the same Affymetrix arrays. We conducted analysis using Partek, Ingenuity Pathway Analysis and GOEAST. Principal Component Analysis (PCA) and hierarchical clustering clearly separated the in vivo from the in vitro groups for both cells types and transcriptomes were more homogeneous upon culture. In both cell cultures behaviours associated with cell adhesion, migration and interaction with matrix or substrate were more abundant. However, the pathways involved generally differed between the two cell types. With the thecal cultures a gene expression signature of an immune response was more abundant, probably by leukocytes amongst the cells cultured from the theca interna. These results indicate differences between in vivo and in vitro that should be considered when interpreting in vitro data.


Sequence determinants of a microtubule tip localization signal (MtLS).
Microtubule plus-end-tracking proteins (+TIPs) specifically localize to the growing plus-ends of microtubules to regulate microtubule dynamics and functions. A large group of +TIPs contain a short linear motif, SXIP, which is essential for them to bind to end-binding proteins (EBs) and target microtubule ends. The SXIP sequence site thus acts as a widespread microtubule tip localization signal (MtLS). Here we have analyzed the sequence-function relationship of a canonical MtLS. Using synthetic peptide arrays on membrane supports, we identified the residue preferences at each amino acid position of the SXIP motif and its surrounding sequence with respect to EB binding. We further developed an assay based on fluorescence polarization to assess the mechanism of the EB-SXIP interaction and to correlate EB binding and microtubule tip tracking of MtLS sequences from different +TIPs. Finally, we investigated the role of phosphorylation in regulating the EB-SXIP interaction. Together, our results define the sequence determinants of a canonical MtLS and provide the experimental data for bioinformatics approaches to carry out genome-wide predictions of novel +TIPs in multiple organisms.


Inhomogeneous membrane receptor diffusion explained by a fractional heteroscedastic time series model.
Single particle tracking experiments have recently uncovered that the motion of cell membrane components can undergo changes of diffusivity as a result of the heterogeneous environment, producing subdiffusion and nonergodic behavior. In this paper, we show that an autoregressive fractionally integrated moving average (ARFIMA) with noise given by generalized autoregressive conditional heteroscedasticity (GARCH) can describe inhomogeneous diffusion in the cell membrane, where the ARFIMA process models anomalous diffusion and the GARCH process explains a fluctuating diffusion parameter.


Evaluation of attenuated strains of Mycoplasma gallisepticum as vaccines in young chickens.
Five trials were conducted to evaluate the virulence and the vaccination efficacy of the F, R, S6, and A5969 strains of Mycoplasma gallisepticum (MG) at different in vitro passage levels. Vaccination was done by eye-drop or aerosol, and efficacy was evaluated in terms of air-sac lesions after aerosol challenge with the R strain of MG. Continuing medium passage of these strains of MG resulted in gradual attenuation. Aerosol vaccination with highly attenuated MG at 21 days of age was more effective in stimulating the development of immunity than was eye-drop vaccination at 7 days, but aerosol vaccination induced more air-sac lesions. Aerosol vaccination with high-passaged F or S6 strain provided good immunity against MG challenge. Back-passage in chickens of the F strain of over 300 medium passages did not substantially increase the virulence, but back-passage of the S6 strain at the 258th passage and F strain at the 11th and 252nd passage did result in increased virulence. Both low- and high-passaged F and S6 strains were virulent for turkey poults, but only the high-passage F strain did not significantly (P less than 0.05) affect body weight.


Modified antisense oligonucleotides directed against tumor necrosis factor receptor type I inhibit tumor necrosis factor alpha-mediated functions.
Tumor necrosis factor alpha (TNF alpha), a polypeptide produced by activated macrophages, is a highly pleiotropic cytokine which elicits inflammatory and immunological reactions. The binding of TNF alpha to tumor necrosis factor receptor type I (TNFRI) is considered the initial step responsible for some of the multiple biological functions mediated by TNF alpha. The role of TNF alpha as an inflammatory mediator through human TNFRI makes TNFRI an attractive target for intervention in both acute and chronic inflammatory diseases. In this study, we have identified partial phosphorothioate oligodeoxyribonucleotides (ODNs) containing C-5 propynyl or hexynyl derivatives of 2'-deoxyuridine which specifically inhibited TNFRI and subsequently inhibited the functions of TNF alpha mediated through TNFRI. The most active ODNs were directed against the 3'-poly adenylation signal site on the TNFRI mRNA, and in a cellular assay, gene-specific antisense inhibition occurred in a dose-dependent fashion at submicromolar concentrations, in the presence of Cellfectin. The inhibition of gene expression correlated with the binding affinity of the ODN for the target mRNA. The ODNs lowered TNFRI protein levels and TNF alpha-mediated functions by specifically reducing levels of TNFRI mRNA. These anti-TNFRI ODNs offer a novel approach for controlling biological functions of TNF alpha and may be useful as human therapeutic agents for treating diseases in which TNF alpha has been implicated.


Assessment of Community Levels of Knowledge About Developmental Dysplasia of the Hip, its Risk Factors, Treatment, and Complications in the Riyadh Region, Saudi Arabia.
Background Developmental dysplasia of the hip (DDH) can involve an immature hip, acetabular dysplasia with or without subluxation, or dislocation of the femoral head. The prevalence of DDH in Saudi Arabia is 10.46 per 1000 live births, highlighting the importance of community knowledge of DDH risk factors and treatment to facilitate early diagnosis, treatment, and avoiding complications. Aim Our goal is to assess community awareness of DDH and the knowledge of its risk factors, treatment, and complications among the population in the Riyadh Region, Saudi Arabia. Methods A cross-sectional study on a sample of 412 participants of the general population in the region of Riyadh, Saudi Arabia was conducted using the snowball sampling technique of sending invitations via social media. All statistical analyses were conducted using SPSS Statistics v.23 (IBM Corp., Armonk, NY). Results Our results revealed that out of the 412 participants almost half reported never hearing about DDH (45.6%). Breech presentation, family history, and female gender were unknown as risk factors to 63%, 58%, and 63.60% of participants respectively. Around 60% of the participant viewed age as the determining factor for DDH treatment. Additionally, 42.70% of participants reported no knowledge about DDH complications, and 72.8% had a low knowledge level regarding DDH. Significant higher knowledge scores were found in females participants (p = 0.026), participants with higher education level (p = 0.01), healthcare professionals (p < 0.001), parents of children who have been screened (p < 0.001), and participants having a first-degree relative with DDH (p < 0.001). Conclusion Our study revealed that residents of the Riyadh Region were unaware of DDH and their knowledge regarding its risk factors, treatment, and complications was poor. Therefore, it is important to implement DDH awareness campaigns to increase the community's knowledge of all aspects of DDH.


Identification of B-epitopes in the human papillomavirus 18 E7 open reading frame protein.
A panel of murine mAb raised against a MS2 replicase/HPV 18 E7 fusion protein included 23 reactive by ELISA with HPV 18 E7 determinants. A total of 19 of the 23 recognized linear epitopes in the N-terminal region of the E7 molecule, while the other four were deduced by binding inhibition assays to recognize conformational determinants in this region. All tested antibodies precipitated a 14-kDa peptide doublet that corresponded with the predicted size of the E7 protein, from HeLa cells, but not from HPV 16 E7 containing CaSki cells. HPV 18 E7 protein was detected by immunolabeling with electron microscopy in both the nucleus and the cytoplasm of HeLa cells with the greater proportion occurring in the cytoplasm. No antibody reacted specifically by indirect immunofluorescence with HeLa cells. Weak cross-reactivity of some mAb with the E6 MS2-replicase fusion protein of HPV 16 was detected by ELISA, but no protein of the appropriate size was immunoprecipitated from CaSki cells. It is concluded that the B cell epitopes on the HPV 18 E7 transforming protein are located in the N-terminal region of the molecule and that some are weakly cross-reactive with HPV 16 E6 protein. E7 protein is either present in HeLa cells at a concentration too low to be detected by indirect immunofluorescence, or the N-terminal epitopes are masked by protein conformation or interaction with cellular or other viral components.


Line sepsis in home parenteral nutrition patients: are there socioeconomic risk factors? A Canadian study.
Line sepsis complicates home parenteral nutrition (HPN). This study examined nonmedical risk factors that may contribute to line sepsis and compared 2 HPN programs with different administrative structures (Ontario and British Columbia [BC]) in terms of line sepsis and patient satisfaction. A survey was developed to evaluate possible correlation between line sepsis and (1) patients' perceptions of HPN care, (2) family support, (3) community support, and (4) socioeconomic status. Data were analyzed by categorizing into high- and low-risk groups using a cutoff point. A second method analyzed the incidences of line sepsis as a continuous variable. Sixty-eight patients responded to the survey: 33 from Ontario (62%), 35 from BC (44%). Community agency, socioeconomic and educational status were not significant in determining line sepsis. Patients who had (1) medication or blood work done through the catheter, (2) a higher number of dependents, or (3) had a trained family member involved in HPN were in the high-risk category for line sepsis, in addition to patients who were part-time students or receiving social assistance. When comparing the provinces, there was no difference in line sepsis. However, significant differences between the provinces include (1) BC patients rate their level of care lower; (2) Ontario patients rely more on family members for HPN; and (3) Ontario patients have more community support. Line sepsis may be increased by some nonmedical risk factors. However, when comparing the 2 programs, rates of line sepsis were not influenced by different administrative structures.


Mapping stress inside living cells by atomic force microscopy in response to environmental stimuli.
The response of cells to environmental stimuli, under either physiological or pathological conditions, plays a key role in determining cell fate toward either adaptive survival or controlled death. The efficiency of such a feedback mechanism is closely related to the most challenging human diseases, including cancer. Since cellular responses are implemented through physical forces exerted on intracellular components, more detailed knowledge of force distribution through modern imaging techniques is needed to ensure a mechanistic understanding of these forces. In this work, we mapped these intracellular forces at a whole-cell scale and with submicron resolution to correlate intracellular force distribution to the cytoskeletal structures. Furthermore, we visualized dynamic mechanical responses of the cells adapting to environmental modulations in situ. Such task was achieved by using an informatics-assisted atomic force microscope (AFM) indentation technique where a key step was Markov-chain Monte Carlo optimization to search for both the models used to fit indentation force-displacement curves and probe geometry descriptors. We demonstrated force dynamics within cytoskeleton, as well as nucleoskeleton in living cells which were subjected to mechanical state modulation: myosin motor inhibition, micro-compression stimulation and geometrical confinement manipulation. Our results highlight the alteration in the intracellular prestress to attenuate environmental stimuli; to involve in cellular survival against mechanical signal-initiated death during cancer growth and metastasis; and to initiate cell migration.


X-ray diffraction and optical properties of a noncentrosymmetric borate CaBiGaB(2)O(7).
Single crystals of the noncentrosymmetric borate CaBiGaB(2)O(7) were synthesized by conventional solid state reaction. The purity of the crystal was checked by x-ray powder diffraction. The optical properties were measured by analyzing the diffuse reflectance data obtained with a Shimadzu UV-3101PC double-beam, double-monochromator spectrophotometer. We find a steep absorption edge confirming its semiconducting nature. The optical band gap obtained by extrapolation of a linearlike absorption edge was roughly 2.9 eV consistent with the observed pale yellow color of the sample. Theoretical calculations based on the structural model built from our measured atomic parameters have been performed using the all-electron full potential linearized augmented plane wave method. The generalized gradient approximation (GGA) of the exchange correlation potential as given by Engel-Vosko GGA is used. The frequency-dependent complex dielectric function was calculated and the origin of some of the spectral peaks is discussed. The linear optical properties show strong uniaxial anisotropy and birefringence that favors large second order susceptibility. Our calculations show that the complex second order nonlinear optical susceptibility tensor chi(322) ((2))(omega) is the dominant component having the largest total Re chi(322) ((2))(0) value of about 1.8 pm/V.


Genotypic approaches to therapy in children: a national active surveillance network (GATC) to study the pharmacogenomics of severe adverse drug reactions in children.
A striking failure of modern medicine is the debilitating and lethal consequences of adverse drug reactions (ADRs), which rank as one of the top 10 leading causes of death and illness in the developed world with direct medical costs of 137-177 billion annually US dollars in the USA. Although many factors influence the effect of medications (e.g., age, organ function, drug interactions), genetic factors account for 20% to 95% of drug response variability and play a significant role in the incidence and severity of ADRs. The field of pharmacogenomics seeks to identify genetic factors responsible for individual differences in drug efficacy and ADRs. Pharmacogenomics has led to several genetic tests that provide clinical dosing recommendations. The Genetic Approaches to Therapy in Children (GATC) project is a national project established in Canada to identify novel predictive genomic markers for severe ADRs in children. An ADR surveillance network has been established in eight of Canada's major children's hospitals, serving up to 75% of all Canadian children. The goal of the project is to identify patients experiencing specific ADRs and matched controls, collect DNA samples, and apply genomics-based technologies to identify ADR-associated genetic markers with the goal of preventing serious ADRs in susceptible children.


Mechanoluminescence of ZnS:Mn phosphors and its correlation to impact energy and contact geometry.
The influence of the impulsive mechanical energy and the contact geometrical parameters on stress-induced light emission of ZnS-based phosphors dispersed in polymeric matrix is investigated using modified impact test equipment. At a low to moderate energy region, luminescent pulses consist of two distinguishable peaks that can be assigned to a loading and unloading sequence, respectively. Transient characteristics are found to be independent of the external excitation. An increment of peak emission intensity with higher impact energy and smaller tip diameter indicates a significant relationship of mechanoluminescence to the geometry of contact. Intense emission deviating from the linearity and an absence of the secondary peak are suggested to be the consequence of fractoluminescence.


The performance of neck circumference and other airway assessment tests for the prediction of difficult intubation in obese parturients undergoing cesarean delivery.
Pregnancy and obesity each increase the chance of difficult tracheal intubation. This study assessed the performance of the neck circumference and other bedside tests in predicting difficult intubation, and aimed to identify the optimal cutoff point of the test(s) with the highest performance. Parturients with a body mass index ≥30kg/m2, undergoing a cesarean delivery employing conventional tracheal intubation, were enrolled. Preoperative neck circumference, sternomental distance and modified Mallampati test were examined. Difficult intubation was defined as intubation difficulty scale score ≥5. Five hundred and seventy parturients of mean body mass index 34.1 (±SD 3.8) kg/m2 were recruited. The incidence of difficult intubation was 3.5%. The performance of the neck circumference test, conducted with the patient sitting or supine, and using the area under the receiver operating characteristic (ROC) curves, was 0.6 (95% CI 0.5 to 0.7) and 0.6 (95% CI 0.4 to 0.7) respectively. The area under the ROC for the modified Mallampati test was 0.6 (95% CI 0.5 to 0.7) and for the sternomental distance test was 0.7 (95% CI 0.6 to 0.8). A sternomental distance less than 14.5cm, and a ratio of neck circumference in the sitting position to sternomental distance of greater than or equal to 2.7, were optimal cutoff points to identify difficult intubation. The neck circumference, the sternomental distance, the modified Mallampati test and the ratio of neck circumference to the sternomental distance show limited performance as screening tests to predict difficult intubation among obese parturients.


Urinary acidification and distal renal tubular acidosis.
Historically, renal tubular acidosis (RTA) has been classified on a clinical basis, without any reference to the underlying disorder. Here we review the normal mechanisms of renal acidification and we identify disorders of specific transporters (genetic, disease-related or drug-induced) that lead to the main categories of distal RTA. We also describe the approach to diagnosis and the current treatment of distal RTA.


ALK-rearranged Renal Cell Carcinoma (RCC): A Report of 2 Cases and Review of the Literature Emphasizing the Distinction Between VCL-ALK and Non-VCL-ALK RCC.
Anaplastic lymphoma kinase (ALK) rearrangement-associated renal cell carcinoma (ALK-rearranged RCC) is a new provisional entity that has been included in the 2016 World Health Organization classification of RCCs. We report 2 cases of ALK-rearranged RCC, 1 with a vinculin-ALK (VCL-ALK) fusion and the other with an EML4-ALK fusion. The VCL-ALK RCC occurred in a 14-year-old girl with sickle cell trait and showed features similar to previously described VCL-ALK RCCs, including medullary epicenter, solid architecture, and polygonal cells with cytoplasmic vacuoles. The EML4-ALK RCC occurred in a 14-year-old boy with no evidence of sickle cell trait and had multiple less-specific growth patterns comprising tubular, solid, and tubulopapillary architectures in the desmoplastic stroma, reminiscent of collecting duct carcinoma. Both tumors demonstrated cytoplasmic and membranous ALK protein expression by immunohistochemistry. Fluorescence in situ hybridization confirmed the ALK gene rearrangements in both cases. On review in the literature, we found that solid architecture and cytoplasmic vacuoles were present significantly more frequently in VCL-ALK RCC than in non-VCL-ALK RCC, supporting the distinctive nature of the former.


Surface-screening mechanisms in ferroelectric thin films and their effect on polarization dynamics and domain structures.
For over 70 years, ferroelectric materials have been one of the central research topics for condensed matter physics and material science, an interest driven both by fundamental science and applications. However, ferroelectric surfaces, the key component of ferroelectric films and nanostructures, still present a significant theoretical and even conceptual challenge. Indeed, stability of ferroelectric phase per se necessitates screening of polarization charge. At surfaces, this can lead to coupling between ferroelectric and semiconducting properties of material, or with surface (electro) chemistry, going well beyond classical models applicable for ferroelectric interfaces. In this review, we summarize recent studies of surface-screening phenomena in ferroelectrics. We provide a brief overview of the historical understanding of the physics of ferroelectric surfaces, and existing theoretical models that both introduce screening mechanisms and explore the relationship between screening and relevant aspects of ferroelectric functionalities starting from phase stability itself. Given that the majority of ferroelectrics exist in multiple-domain states, we focus on local studies of screening phenomena using scanning probe microscopy techniques. We discuss recent studies of static and dynamic phenomena on ferroelectric surfaces, as well as phenomena observed under lateral transport, light, chemical, and pressure stimuli. We also note that the need for ionic screening renders polarization switching a coupled physical-electrochemical process and discuss the non-trivial phenomena such as chaotic behavior during domain switching that stem from this.


Amino acid substitutions preserve protein folding by conserving steric and hydrophobicity properties.
We present a comprehensive analysis of amino acid substitution patterns (sets of residues in a position of a multiple alignment) and conservation of physicochemical properties in alignments of protein sequences. Of the one million possible substitution patterns, only a few hundred account for the majority of aligned positions. Very similar distributions of substitution patterns are observed in all but one of the diverse databases of multiple alignments. In these substitution patterns we analyzed the conservation of 511 physicochemical and steric amino acid properties. Highest conservation was observed in those steric and transfer free energy-related properties that are crucial for folding. The best conserved steric properties include the minimal width of the side chains and their interactions with other residues. Among the hydrophobicity-related properties, charge and those properties that provide information on propensities to form secondary structures or side chain conformation, appear to be better conserved than pure hydrophobicity measures. Physicochemical sequence analysis based on the most conserved properties is expected to aid searching a protein sequence query against a database of multiple alignments, prediction of secondary and tertiary structures and protein engineering.


IL-17-induced Act1-mediated signaling is critical for cuprizone-induced demyelination.
Cuprizone inhibits mitochondrial function and induces demyelination in the corpus callosum, which resembles pattern III lesions in multiple sclerosis patients. However, the molecular and cellular mechanism by which cuprizone induces demyelination remains unclear. Interleukin-17 (IL-17) secreted by T helper 17 cells and γδT cells are essential in the development of experimental autoimmune encephalomyelitis. In this study, we examined the importance of IL-17 signaling in cuprizone-induced demyelination. We found that mice deficient in IL-17A, IL-17 receptor C (IL-17RC), and adaptor protein Act1 (of IL-17R) all had reduced demyelination accompanied by lessened microglial and polydendrocyte cellular reactivity compared with that in wild-type mice in response to cuprizone feeding, demonstrating the essential role of IL-17-induced Act1-mediated signaling in cuprizone-induced demyelination. Importantly, specific deletion of Act1 in astrocytes reduced the severity of tissue injury in this model, indicating the critical role of CNS resident cells in the pathogenesis of cuprizone-induced demyelination. In cuprizone-fed mice, IL-17 was produced by CNS CD3(+) T cells, suggesting a source of IL-17 in CNS upon cuprizone treatment.


Relationship of histologic grade of hepatocellular carcinoma (HCC) to tumor size, and demonstration of tumor cells of multiple different grades in single small HCC.
The histologic features of 65 surgically resected cases of hepatocellular carcinoma (HCC) were studied, and the cellular differentiation was graded from I to IV according to Edmondson-Steiner's classification. HCC cells of 2 or more grades were seen in 31 (47.7%) of the 65 cases. There was no significant difference in the proportion of HCC composed of tumor cells of more than one histologic grade between HCCs with association with liver cirrhosis and without cirrhosis. Among these 31 cases, extremely well-differentiated HCC, corresponding to Edmondson-Steiner's grade I carcinoma, was found in 9 of 11 tumors smaller than 3 cm in diameter, but it was not seen in 20 tumors larger than 3 cm in diameter. In the 34 HCCs with a uniform histologic pattern, all of two minute tumors smaller than 1 cm in diameter consisted of extremely well-differentiated HCC, but there were no cases consisting of extremely well-differentiated HCC in tumors larger than 2 cm in diameter. Taken together, these findings suggest that HCCs originate as relatively well-differentiated tumors, which may be difficult to distinguish from adenomatous regenerative nodules, and become progressively less differentiated at a later stage of their development.


A quantitative stereological description of the ultrastructure of normal rat liver parenchymal cells.
The principles of stereology have been applied to a morphometric analysis of parenchymal cells from the peripheral, midzonal, and central regions of normal rat liver lobules. The fractional volumes of cytoplasm occupied by mitochondria, peroxisomes, lysosomes, lipid, and glycogen have been determined. The surface densities of smooth- and rough-surfaced endoplasmic reticulum and of mitochondrial envelope and cristae have also been measured. The average number and dimensions of mitochondria and peroxisomes have been evaluated. By the use of an independent measurement of the average cytoplasmic volume, these data have been expressed as the actual volumes, areas, and numbers per cell in the different parts of the hepatic lobule. Similarly, the volumes of the envelope, cristae, and matrix compartments and the area of cristae membranes have been calculated for the average-sized mitochondrion in each lobular zone. Structural homogeneity is found in over 80% of normal rat liver parenchymal cells, with most of the significant differences being confined to those cells immediately surrounding the central veins.


Effects of elevated plasma epinephrine on glucose utilization and blood flow in conscious rat brain.
Acute glucoprivation increases cerebral blood flow (CBF), which is often attributed to the associated rise in plasma epinephrine levels. This study examined directly the effects of comparable increases in plasma epinephrine levels achieved by continuous intravenous infusions of epinephrine in normoglycemic, unanesthetized rats on local and overall CBF and cerebral glucose utilization (1CMRglc). CBF was determined by the autoradiographic [14C]iodoantipyrine method in six unanesthetized rats in which epinephrine dissolved in 1% ascorbic acid-1 mM EDTA was infused at a rate of 1 microg/min and in five normal controls infused with the vehicle alone. 1CMRglc was determined by the autoradiographic [14C]deoxyglucose method in six conscious rats infused similarly with the epinephrine solution and in six normal controls treated with the vehicle alone. The epinephrine infusions raised arterial plasma epinephrine levels 10- to 20-fold and increased arterial blood pressure and plasma glucose levels. Local CBF, however, was significantly changed (P < 0.05, Student's t-test) in only 2 of 25 structures examined, and the changes were decreases not increases. 1CMRglc was not changed significantly in any of 42 brain structures examined, and average blood flow and glucose utilization in the brain as a whole were unaffected. These results show that high circulating levels of epinephrine similar to those accompanying glucoprivation alter neither local nor overall CBF and glucose utilization and cannot explain the increases in CBF associated with glucoprivation.


Salivary glands in long-term alloxan-diabetic rats. A quantitative light and electron-microscopic study.
Fifty untreated diabetic animals were compared with 58 age-matched non-diabetic controls. Reduced salivary gland weight was evident after one month's diabetes and this was unchanged after 12 months of diabetes. Submandibular/sublingual gland weight was proportional to the reduced body weight in the diabetic rats. Parotid gland weight, however, was proportionally more reduced. Only diabetic rats had lipid inclusions in the acinar cells of their submandibular glands and the morphometrically estimated amount of inclusions was positively correlated to the blood glucose level. Acinar cell size was significantly increased in long-term diabetic rats as compared with short-term diabetic rats and controls. Capillary basement membrane width was significantly increased in long-term diabetic rats compared with age-matched controls and with short-term diabetic rats. Thus, both the degree and duration of diabetes have a major effect on salivary gland morphology in alloxan diabetic rats.


The effects on gastric emptying and carbohydrate loading of an oral nutritional supplement and an oral rehydration solution: a crossover study with magnetic resonance imaging.
Preoperative administration of clear fluids by mouth has recently been endorsed as a way to improve postoperative outcomes. A carbohydrate-containing beverage supplemented with electrolytes or proteins may have additional benefits for patients' satisfaction. However, effects on gastric residual, nausea, and emesis and the effectiveness of these beverages for improving patients' hydration status have not been well defined. We evaluated changes in gastric volume over time by magnetic resonance imaging, as well as blood glucose levels, before and after administration of 500 mL oral rehydration solution (ORS) containing 1.8% glucose and electrolytes in 10 healthy volunteers. The same volume of an oral nutritional supplement (ONS) containing 18% glucose and supplemental arginine (545 mOsm/kg) was given to the same population using a crossover design. The mean (median, 95% confidence interval) gastric fluid volume at 1 hour after oral ingestion was 55.0 (55.3, 39.0-70.9) mL in the ORS group, whereas 409.2 (410.9, 371.4-447.0) mL in the ONS group (P = 0.0002). The gastric fluid volume of all participants in the ORS group returned to <1 mL/kg at 90 minutes after ingestion, whereas none reached <1 mL/kg at 120 minutes in the ONS group. The ONS group showed a sustained increase in the blood glucose level after ingestion (P < 0.0001 to baseline at 30, 60, 120 minutes), while the ORS group showed an initial increase (P < 0.0001, P = 0.01, P = 0.205 at each time point). ORS supplemented with a small amount of glucose showed faster gastric emptying, which may make it suitable for preoperative administration. In contrast, ONS supplemented with arginine with a relatively low osmolality was associated with a longer time for gastric emptying, although it showed a sustained increase in blood glucose level.


Constructing, Perceiving, and Maintaining Scenes: Hippocampal Activity and Connectivity.
In recent years, evidence has accumulated to suggest the hippocampus plays a role beyond memory. A strong hippocampal response to scenes has been noted, and patients with bilateral hippocampal damage cannot vividly recall scenes from their past or construct scenes in their imagination. There is debate about whether the hippocampus is involved in the online processing of scenes independent of memory. Here, we investigated the hippocampal response to visually perceiving scenes, constructing scenes in the imagination, and maintaining scenes in working memory. We found extensive hippocampal activation for perceiving scenes, and a circumscribed area of anterior medial hippocampus common to perception and construction. There was significantly less hippocampal activity for maintaining scenes in working memory. We also explored the functional connectivity of the anterior medial hippocampus and found significantly stronger connectivity with a distributed set of brain areas during scene construction compared with scene perception. These results increase our knowledge of the hippocampus by identifying a subregion commonly engaged by scenes, whether perceived or constructed, by separating scene construction from working memory, and by revealing the functional network underlying scene construction, offering new insights into why patients with hippocampal lesions cannot construct scenes.


Genetic signature for human risk assessment: lessons from trichloroethylene.
Trichloroethylene (TCE), an organic solvent commonly used for metal degreasing and as a chemical additive, is a significant environmental contaminant that poses health concerns in humans. The US Environmental Protection Agency (EPA) is currently revising the 2001 TCE human risk assessment draft. The next draft is expected to be ready in 2008. TCE metabolites are detectable in humans and carry varying potencies for induction of cancers in animals. Genomic mechanisms have been explored in animals and humans to link TCE to carcinogenesis. DNA analysis provides an opportunity for detection of unique genetic alterations representing a signature of TCE exposure. These alterations can arise from genotoxic and nongenotoxic pathways at multiple points throughout tumorigenesis. Although fixation of alterations may require several stages of selection and modification, the spectra can be specific to TCE. Only a fraction of these alterations eventually lead to tumor formation and some contribute to tumor progression. Genetic events in two major TCE target organs are reviewed, including the VHL gene in kidney, and the Ras gene and genome-wide hypomethylation in liver. Attempts to identify a genetic signature of TCE exposure are challenged by inconsistent findings, lack of evidence of promutagenic lesions, biological relevance of specific genomic changes, and likelihood of coexposures. For human risk assessment, genome-wide screening is useful and is possible with the development of new DNA-sequencing technologies. Genetic screening for preneoplastic and tumor tissues from high-risk population is proposed to exclude the noise of passenger mutations and genetic polymorphisms.


Plasma carcinoembryonic antigen in the diagnosis and management of patients with hepatocellular carcinoma.
The value of serial carcinoembryonic antigen (CEA) measurements as a marker of disease progression or in monitoring treatment was investigated in patients with hepatocellular carcinoma. Of 40 patients, including 16 with normal serum alpha-fetoprotein (AFP) concentrations, 29 (72.5%) had abnormal plasma CEA at presentation. Although this was more common in patients with pre-existing cirrhosis, the mean and range of plasma CEA were similar in patients with and without pre-existing hepatic disease. There was no correlation between plasma CEA and any biochemical parameter of hepatic function, although plasma CEA concentrations were significantly lower in patients with well-differentiated tumors. CEA concentrations increased in 71% of patients who had no response to cytotoxic drugs, but CEA also increased in 62.5% of those patients who did respond. Plasma CEA concentrations were elevated in 62.5% of patients with normal and 79% of patients with raised serum AFP on admission to the hospital. There was no correlation between individual AFP and CEA concentrations. Although elevated plasma CEA levels may be of diagnostic value in patients with hepatocellular carcinoma in the absence of pre-existing hepatic disease, and in those with normal serum AFP, our findings indicate that it does not behave as a true tumor marker.


Being with feelings as a recognition practice: developing clients' self-understanding.
How are shifts in clients' self-understanding related to focusing on feelings during therapy sessions? A phenomenological study involving matched-pairs of clients and nurse-therapists using video-cued narrative reflection. Feelings disclose significance within a therapy session. Dialogical relationships and involvements help clients experience the feelings they are discussing, obtain feedback from therapists, and develop self-understanding. Nurse educators should stress the importance of focusing on feelings in a session, and clinicians should practice this skill to facilitate a client's developing self-understanding.


Clinical Impact of Tumor Mutational Burden in Neuroblastoma.
Neuroblastoma is the most common pediatric extracranial solid tumor. Within conventional risk groups, there is considerable heterogeneity in outcomes, indicating the need for improved risk stratification. In this study we analyzed the somatic mutational burden of 515 primary, untreated neuroblastoma tumors from three independent cohorts. Mutations in coding regions were determined by whole-exome/genome sequencing of tumor samples compared to matched blood leukocytes. Survival data for 459 patients were available for analysis of 5-year overall survival using the Kaplan-Meier method and log-rank test. All statistical tests were two-sided. Despite a low overall somatic mutational burden (mean = 3, range = 0-56), 107 patients were considered to have high mutational burden (>3 mutations). Unfavorable histology and age 18 months and older were associated with high mutational burden. Patients with high mutational burden had inferior 5-year overall survival (29.0%, 95% confidence interval [CI] = 17.2 to 41.8%) vs those with three or fewer somatic mutations (76.2%, 95% CI = 71.5 to 80.3%) (log-rank P < .001) and this association persisted when limiting the analysis to genes included on a 447-gene panel commonly used in clinical practice. On multivariable analysis, mutational burden remained prognostic independent of age, stage, histology and MYCN status. This study demonstrates that mutational burden of primary neuroblastoma may be useful in combination with conventional risk factors to optimize risk stratification and guide treatment decisions, pending prospective validation.


Revascularization in a 17-Year-Old Girl with Neurofibromatosis and Severe Hypertension Caused by Renal Artery Stenosis.
Renal artery stenosis caused by neurofibromatosis is a rare cause of renovascular hypertension. This hypertension can develop during childhood and is one of the leading causes of poor outcome. We report the case of a 17-year-old girl who was incidentally diagnosed with severe hypertension. During her examination for secondary hypertension, we reached a diagnosis of neurofibromatosis type 1 on the basis of a cluster of typical findings: optic nerve glioma, café au lait spots, nodular neurofibromas, and axillary freckling. Renal angiograms revealed a hemodynamically significant left renal artery stenosis (70%). Renal angioplasty with a self-expanding stent was performed one month later for rapidly progressive renal artery stenosis (90%) and uncontrolled blood pressure. Excellent blood pressure control resulted immediately and was maintained as of the 2-year follow-up evaluation. We think that percutaneous transluminal renal angioplasty can be effective in select patients who have neurofibromatosis type 1 and refractory hypertension caused by renal artery stenosis.