Neurodegeneration-Like Pathological and Behavioral Changes in an AAV9-Mediated p25 Overexpression Mouse Model.
The transgenic mice models overexpressing human p25 contribute greatly to the in vivo neurotoxic mechanism of p25 in neurodegenerative diseases. However, it is time-consuming to manipulate existing transgenic mice models. Here we aim to establish a novel mouse model of neurodegeneration by overexpressing p25 mediated by recombinant adeno-associated virus serotype 9 (rAAV9). AAV9-GFP-p25 encoding GFP-fused p25 driven by synapsin promoter, and the control, AAV9-GFP, were delivered in mice by tail-vein injection. Assessments of p25 expression, neurodegenerative pathology, and behavioral changes were performed. GFP expression was detected by in vivo imaging as early as one week after virus injection. Notably, widespread expression of p25 was obviously found in cortex, hippocampus, and cerebellum in AAV9-GFP-p25 mice. Moreover, decreased hippocampus volumes in AAV9-GFP-p25 mice were detected by 7T MRI examination about one month after injection. Further, these AAV9-GFP-p25 mice exhibited progressive memory impairment from three-month to six-month after virus injection. At last, hyperphosphorylated tau, neurofibrillary tangles, activated astrocytes and microglia cells were elevated in these p25 mice at about six months after virus delivery. However, amyloid-β plaques, overt neuronal loss, and apoptosis in the hippocampus and cortex were not significantly induced by AAV9-mediated p25 overexpression. The AAV9-mediated p25 overexpression mouse model, which is a practical model exhibiting neurodegeneration-like pathological and behavioral changes, provides an easier and time-saving method to explore the functions of p25 in vivo, as well as an alternative tool for development of drugs against neurotoxic of p25.


Methyl 3-amino-2,3,6,-trideoxy-L-hexopyranosides in DFT level theory conformational studies.
Geometry optimizations at the B3LYP level of density functional theory are reported for the (1)C4 and (4)C1 conformations of four theoretically possible alpha and beta methyl 3-amino-2,3,6-trideoxy-L-hexopyranosides. The Gibbs free energies, relative Gibbs free energies, and geometry parameters are presented for all the optimized structures. Conformational analysis of the pyranose ring is performed for each stereoisomer on the basis of calculated rotamer populations. It is demonstrated that the alpha/beta-L-arabino, alpha/beta-L-lyxo, and alpha-L-ribo stereoisomers adopt the (1)C4 conformation, whereas beta-L-ribo and alpha/beta-L-xylo stereoisomers remain in (1)C4 <==> (4)C1 conformational equilibrium. The preference of the alpha over the beta anomers is due to the endo-anomeric effect. The factors affecting the stability of pyranose ring conformations are discussed, as is the influence of hydrogen bonds on the orientation of the hydroxyl and amino groups. Figures of the most stable conformers are presented.


Serum Lipid Abnormalities and Nonalcoholic Fatty Liver Disease in Adult Males.
Dyslipidemia is a risk factor for nonalcoholic fatty liver disease (NAFLD). The aim of our study was to determine the associations of serum lipid indexes with NAFLD in adult males. In this cross-sectional study, 830 patients with NAFLD and 2,357 healthy individuals were assessed. Serum total cholesterol (TC), triglyceride (TG), high-density lipoprotein cholesterol and low-density lipoprotein cholesterol (LDL-C) levels were compared between patients with NAFLD and controls. The associations of dyslipidemia indexes with NAFLD occurrence were assessed by univariate analysis, and multivariate analysis was performed to identify independent dyslipidemia factors predictive of NAFLD. Of the 3,187 study subjects, NAFLD occurred in 830 (26.04%), there were 504 (60.72%) patients with mild disease and 326 (39.28%) patients with moderate-to-severe disease. Although the frequency of normal TC, TG, LDL-C and high-density lipoprotein cholesterol levels in patients with NAFLD was similar to the controls, the frequencies of patients with NAFLD with marginally high and high TC, TG and LDL-C levels were significantly different when compared with controls. Interestingly, the association of the number of abnormal serum lipid indexes and NAFLD was highly significant with 2 abnormalities (odds ratio = 1.977; 95% CI: 1.436-2.722; P < 0.001) and ≥3 abnormalities (odds ratio = 3.505; 95% CI: 2.466-4.982; P < 0.001). A significant positive association was found between dyslipidemia characteristics and NAFLD in adult males.


Early Pup Removal Leads to Social Dysfunction and Dopamine Deficit in Late Postpartum Rats: Prevention by Social Support.
Offspring interaction is among the most highly motivated behaviors in maternal mammals and is mediated by mesolimbic dopamine (DA) system activation. Disruption or loss of significant social relationships is among the strongest individual predictors of affective dysregulation and depression onset in humans. However, little is known regarding the effects of disrupted mother-infant attachment (pup removal) in rat dams. Here, we tested the effects of permanent pup removal in rat dams, which were assigned to one of three groups on postpartum day (PD) 1: pups; pups removed, single-housed; or pups removed, co-housed with another dam who also had pups removed; and underwent a behavioral test battery during PD 21-23. In vivo electrophysiological recordings of ventral tegmental area (VTA) DA neurons were performed on PD 22 and 23 in a subset of animals. Pup removal did not impact sucrose consumption or anxiety-like behavior, but increased passive forced swim test (FST) coping responses. Pup-removal effects on social behavior and VTA activity were sensitive to social buffering: only single-housed dams exhibited reduced social motivation and decreased numbers of active DA neurons. Dams that had pups removed and were co-housed did not exhibit changes in social behavior or VTA function. Moreover, no changes in social behavior, FST coping, or VTA activity were found in socially isolated adult virgin females, indicating that effects observed in dams are specific to pup loss. This study showed that deprivation of species-expected social relationships (pups) during the postpartum precipitates an enduring negative affect state (enhanced passive coping, blunted social motivation) and attenuated VTA DA function in the dam, and that a subset of these effects is partially ameliorated through social buffering.


The Roles of Exosomes in Visual and Auditory Systems.
Exosomes are nanoscale membrane-enclosed vesicles 30-150 nm in diameter that are originated from a number of type cells by the endocytic pathway and consist of proteins, lipids, RNA, and DNA. Although, exosomes were initially considered to be cellular waste, they have gradually been recognized to join in cell-cell communication and cell signal transmission. In addition, exosomal contents can be applied as biomarkers for clinical judgment and exosomes can as potential carriers in a novel drug delivery system. Unfortunately, purification methods of exosomes remain an obstacle. We described some common purification methods and highlight Morpho Menelaus (M. Menelaus) butterfly wings can be developed as efficient methods for exosome isolation. Furthermore, the current research on exosomes mainly focused on their roles in cancer, while related studies on exosomes in the visual and auditory systems are limited. Here we reviewed the biogenesis and contents of exosomes. And more importantly, we summarized the roles of exosomes and provided prospective for exosome research in the visual and auditory systems.


Mulitmodal Corneal Imaging of Genetically Confirmed Keratitis-Ichthyosis-Deafness Syndrome.
Keratitis-ichthyosis-deafness (KID) syndrome is characterized by a congenital triad of keratitis, ichthyosis, and deafness, and is most commonly associated with mutations in the gap junction protein beta 2 gene (GJB2) on chromosome 13q11-q12. Multimodal anterior segment imaging and genetic testing were used to supplement clinical examination findings in the diagnosis and management of a 12-year-old boy with suspected KID syndrome. The patient presented with hearing loss, ichthyosis of the face and extremities, and corneal scarring and keratinization. The corneal limbal stem cell population was found to be normal on in vivo confocal microscopy, whereas the basal epithelium of the cornea demonstrated scarring and areas of cellular loss. Screening of GJB2 revealed a presumed pathogenic heterozygous missense mutation, c.148G>A, confirming the diagnosis of KID syndrome. Multimodal imaging including in vivo confocal microscopy suggests that dysfunctional corneal basal epithelium maturation might contribute to the pathophysiology of keratopathy in KID syndrome.


Providing palliative care in the ambulatory care setting.
Palliative care that provides specialized attention to pain and symptom management is important for patients with cancer. Palliative care aims to reduce pain and other symptoms through an interdisciplinary approach involving physicians, nurses, social workers, and other members of the healthcare team. Families are included in care planning. Patients and families benefit from the availability of palliative care services early in the disease process, particularly when symptoms impact quality of life. One way to implement early palliative interventions is the establishment of an ambulatory care clinic dedicated to palliative care. This article describes the experience of an outpatient palliative care clinic at a large teaching hospital by using case studies to highlight the benefits of ambulatory palliative care and concluding with recommendations for research.


First record of the Asian ambrosia beetle, Xylosandrus crassiusculus (Motschulsky) (Coleoptera: Curculionidae, Scolytinae), in Slovenia.
Xylosandrus crassiusculus (Coleoptera: Curculionidae, Scolytinae), with the common name the Asian ambrosia beetle or the granulate ambrosia beetle, originates in tropical and subtropical regions of eastern Asia. It is one of the most widespread wood-boring beetles and among these one of the most successful invaders (IPPC 2017). Outside its native habitat, the species is present in Africa (Wood Bright 1992, Atkinson et al. 2000), in Australia (IPPC 2017), on the Pacific Islands (Beaver 1976), in the Americas (Atkinson 1988, Rabaglia et al. 2006, Flechtmann Atkinson 2016, Landi et al. 2017) and in Europe (Pennacchio et al. 2003, Nageleisen et al. 2015, Gallego et al. 2016, Francardi et al. 2017).


Non-invasive mid-term electrocardiogram patch monitoring is effective in detecting atrial fibrillation.
Atrial fibrillation (AF) is a cause of serious morbidity such as stroke. Early detection and treatment of AF is important. Current guidelines recommend screening via opportunistic pulse taking or 12‑lead electrocardiogram. Mid-term ECG patch monitors increases the sensitivity of AF detection. The Singapore Atrial Fibrillation Study is a prospective multi-centre study aiming to study the incidence of AF in patients with no prior AF and a CHA2DS2-VASc score of at least 1, with the use of a mid-term continuous ECG monitoring device (Spyder ECG). Consecutive patients from both inpatient and outpatient settings were recruited from 3 major hospitals from May 2016 to December 2019. Three hundred and fifty-five patients were monitored. 6 patients (1.7%) were diagnosed with AF. There were no significant differences in total duration of monitoring between the AF and non-AF group (6.39 ± 3.19 vs 5.42 ± 2.46 days, p = 0.340). Patients with newly detected AF were more likely to have palpitations (50.0% vs 11.8%, p = 0.027). Half of the patients (n = 3, 50.0%) were diagnosed on the first day of monitoring and the rest were diagnosed after 24 h. On univariate analysis, only hyperlipidemia was associated with reduced odds of being diagnosed with AF (OR HR 0.08 CI 0.01-0.74, p = 0.025). In a group of 128 patients who underwent coronary artery bypass grafting and had post-operative ECG monitoring, 9 patients (7.0%) were diagnosed with post-operative AF. The use of non-invasive mid-term patch-based ECG monitoring is an effective modality for AF screening.


Structure-property relationships in porous 3D nanostructures: epoxy-cross-linked silica aerogels produced using ethanol as the solvent.
Cross-linking silica aerogels with organic groups has been shown to improve the strength over un-cross-linked aerogels by as much as 2 orders of magnitude. Previous cross-linking chemistry has been developed using solvents specifically chosen to dissolve the monomers and accommodate the reaction temperature. Because the process of making the aerogels requires so much solvent, it is of interest to consider less toxic solvents such as ethanol to increase safety and enhance scale up. To this end, two different epoxy precursors with suitable solubility in ethanol were evaluated as cross-linkers for silica gels prepared from (3-aminopropyl)triethoxysilane and tetraethylorthosilicate. In addition, 1,6-bis(trimethoxysilyl)hexane (BTMSH) was used as an additive in the underlying silica structure to add flexibility to the aerogels. It was found that the ethanol-derived aerogels exhibited more shrinkage than those prepared from other solvents but that including BTMSH in the aerogels significantly reduced this shrinkage. Inclusion of BTMSH also imparted the ability of the aerogel monoliths to recover elastically when compressed up to 50% strain. In addition, optimized cross-linked aerogels prepared in this study have mechanical properties comparable to those using other more undesirable solvents and cross-linkers.


Lysosomal glycerophosphocholine phosphodiesterase in Tetrahymena.
The purification and characterization of a novel phosphodiesterase (PDE) is presented. The activity was detected in the extracellular medium of Tetrahymena thermophila cultures, by the release of p-nitrophenol from p-nitrophenylphosphocholine (PNPPC) with an acidic pH optimum. In cell homogenates, it is sedimentable, shows a latency similar to that of acid phosphatase and is co-secreted with this enzyme, indicating that it is a lysosomal hydrolase. PNPPC-PDE was purified to homogeneity from the extracellular medium, yielding a single band of 58 kD on SDS polyacrylamide gel electrophoresis. It catalyzed the release of glycerol from glycerophosphocholine (GPC) and GPC competitively inhibits degradation of PNPPC. We present further evidence indicating that the natural substrate for PNPPC-PDE is GPC. Thus, Tetrahymena becomes the first eukaryote in which a lysosomal GPC-PDE is observed. This finding provides a new pathway for the complete breakdown of phosphatidylcholine in a lysosomal medium.


Neuromas of the extrahepatic bile ducts as a cause of obstructive jaundice.
We present four cases of extrahepatic bile duct neuromas causing obstructive jaundice. Two patients with neuromas localized in the hepatic duct had no evidence of gallstones or history of previous surgery. In these subjects, partial resection of the common bile duct and cholecystectomy was followed by hepatico-jejunostomy with Roux-en-Y reconstruction. In another patient, 7 years after cholecystectomy, a neuroma localized in the hepatic duct was treated by insertion of a T-tube. In the fourth patient, 9 years after laparoscopic cholecystectomy, partial common bile duct resection and Roux-en-Y hepatico-jejunostomy was performed. In all cases, the correct diagnosis of this very rare cause of extrahepatic bile duct obstruction was established postoperatively.


Allometric conservatism in the evolution of bird beaks.
Evolution can involve periods of rapid divergent adaptation and expansion in the range of diversity, but evolution can also be relatively conservative over certain timescales due to functional, genetic-developmental, and ecological constraints. One way in which evolution may be conservative is in terms of allometry, the scaling relationship between the traits of organisms and body size. Here, we investigate patterns of allometric conservatism in the evolution of bird beaks with beak size and body size data for a representative sample of over 5000 extant bird species within a phylogenetic framework. We identify clades in which the allometric relationship between beak size and body size has remained relatively conserved across species over millions to tens of millions of years. We find that allometric conservatism is nonetheless punctuated by occasional shifts in the slopes and intercepts of allometric relationships. A steady accumulation of such shifts through time has given rise to the tremendous diversity of beak size relative to body size across birds today. Our findings are consistent with the Simpsonian vision of macroevolution, with evolutionary conservatism being the rule but with occasional shifts to new adaptive zones.


Space maintainer effects on intercanine arch width and length.
In order to investigate the effects of space maintainers in intercanine arch width and length, twenty cases, characterized with the early loss of mandibular primary molars were selected and divided into two groups. The treatment group used removable space maintainers, while the other ten cases served as the control group. The first dental casts of the treatment and control groups were obtained when the primary canines were in the mouth. After the eruption of permanent canines second dental casts were obtained in both groups. Six measurements were made on the dental casts of each patient. No parameter was found to be statistically significant in the treatment group. In the control group the increase in intercanine arch width and perimeter were found to be statistically significant. Also the increase at the buccal and lingual bone measurements were found to be statistically significant. These results showed that space maintainers might cease the increase in intercanine arch width and length during the transition period between the primary and permanent canines.


Demographic and clinical characteristics of pediatric COVID-19 in Arkansas: March-December 2020.
The COVID-19 pandemic reached the United States in early 2020 and spread rapidly across the country. This retrospective study describes the demographic and clinical characteristics of 308 children presenting to an Arkansas Children's emergency department (ED) or admitted to an Arkansas Children's hospital with COVID-19 in the first 10 months of the COVID-19 pandemic, prior to the emergence of clinically significant variants and available vaccinations. Adolescents aged 13 and older represented the largest proportion of this population. The most common presenting symptoms were fever, gastrointestinal symptoms, and upper respiratory symptoms. Patients with multisystem inflammatory syndrome in children (MIS-C) had a longer length of stay (LOS) than patients with acute COVID-19. Children from urban zip codes had lower odds of admission but were more likely to be readmitted after discharge. Nearly twenty percent of the study population incidentally tested positive for COVID-19. Despite lower mortality in children with COVID than in adults, morbidity and resource utilization are significant. With many Arkansas children living in rural areas and therefore far from pediatric hospitals, community hospitals should be prepared to evaluate children presenting with COVID-19 and to determine which children warrant transport to pediatric-specific facilities.


Acute heart failure syndromes and coronary perfusion.
Acute heart failure syndromes (AHFS), with a high post-discharge mortality and rehospitalization rate, represent a significant public health burden. The treatment of patients hospitalized with AHFS often includes the use of vasoactive medications such as inotropes and vasodilators. Although such agents are frequently used, their safety and efficacy remain controversial. A significant number of patients with heart failure have underlying coronary artery disease and may be at greater risk from hemodynamic alterations that can diminish coronary perfusion. In AHFS, the relationship among vasoactive medications, coronary perfusion, and potential myocardial injury needs further investigation. Newer techniques now available to evaluate coronary perfusion should provide guidance for the evaluation of existing and future vasoactive therapies for AHFS.


Redefining cutaneous lymphatic flow: the necessity of preoperative lymphoscintigraphy in the management of malignant melanoma.
The purpose of this study is to emphasize the instrumental role of preoperative lymphoscintigraphy in the surgical treatment of patients with malignant melanoma. The efficacy of lymphoscintigraphy is reflected in its ability to reveal cutaneous lymphatic drainage to regional nodal basins that are at risk for melanoma metastases but not necessarily discernable to be at risk through standard historical anatomical guidelines or clinical experience. This preoperative lymphatic mapping technique has contributed greatly to the accuracy and efficiency of staging procedures including sentinel node biopsy and elective lymph node dissection. After informed consent, a selected series of four patients with primary melanomas located in watershed areas of the body (left neck, right mid-abdomen, right scapula, left back) and two patients with extremity melanomas (right distal forearm and left ankle) underwent pre-operative lymphoscintigraphy to identify all basins for metastases. In all of the cases, lymphatic drainage occurred in an unusual and unexpected basin that could not have been predicted clinically and in three of the cases the resected basins contained positive sentinel nodes. If not for the preoperative lymphoscintigraphy, these nodal basins would not have been resected and metastatic disease would have been left behind. In addition, the staging of the melanoma patient would have been inaccurate. If the sentinel node biopsy of elective lymph node dissection (ELND) were based on clinical predictions only, nodes equally at risk for metastatic disease would not have been resected and in some cases, nodal basins not at risk for metastases would have been resected unnecessarily. Without lymphoscintigraphy, the validity and efficacy of the ELND or the sentinel node biopsy for nodal staging is greatly compromised. These six case studies illustrate the difficulty of predicating lymphatic drainage from primary sites located on the head and neck, truck and even the extremities and demonstrate the indispensability of preoperative lymphoscintigraphy in the management of malignant melanoma.


Randomized, controlled comparison of two forms of preparation for screening flexible sigmoidoscopy.
There is a paucity of data regarding the optimal form of bowel preparation for flexible sigmoidoscopy. Most endoscopists recommend enemas. A simpler preparation that is easy, acceptable, and that reduces patient encounter time would be desirable, and might be cost-effective. Our objective in this study was to evaluate a simple oral form of preparation for screening flexible sigmoidoscopy. In this randomized, single-blind, controlled trial, we compared two forms of preparation in consecutive male patients referred for screening flexible sigmoidoscopy. The oral preparation consisted of one bottle of magnesium citrate and two "Dulcolax" tablets on the evening before flexible sigmoidoscopy. This was compared with the standard form of preparation, namely, two Fleet's enemas given on arrival at the endoscopy suite. Thirty-seven patients received the oral preparation [mean age, 62.8 +/- 8.9 (SD) yr]; 33 received enemas (mean age, 65.2 +/- 7.3 yr). Endoscopists were blinded to the preparation. Mean time between arrival and starting flexible sigmoidoscopy was 36 +/- 22 (SD) min for patients on oral preparation, and 62 +/- 25 min for patients receiving enemas (p < 0.0001). Mean times performing flexible sigmoidoscopy were 10 +/- 3 min and 13 +/- 4 min, respectively (p = 0.004). Mean patient satisfaction score (range 0-13) was higher for patients given the oral preparation (11.4 +/- 1.8) than for patients receiving enemas (9.6 +/- 2.4) (p = 0.001). Fifteen patients randomized to receive the oral preparation had previous flexible sigmoidoscopy with an enema preparation; all preferred the oral form. Mean technical difficulty (range 1-10) was 3 +/- 2.2 for patients given the oral preparation and 4.9 +/- 3.1 for patients receiving the enema preparation (p = 0.01). Polyps were identified in 10/37 patients who received the oral preparation and in 3/33 patients who received enemas (p = 0.05). Quality of colon preparation was judged "good" in 29, "fair" in four, and "poor" in four, among the 37 patients given the oral form; corresponding values for 33 patients given enemas were 16, 10, and 7 (p = 0.03). Patient acceptance, encounter time, technical ease, and quality of colon preparation were significantly better with the oral form of colon preparation than with the standard Fleet enema preparation.


In vitro and in vivo study of CJY, an isoflavone, on p-glycoprotein function in rats.
The p-glycoprotein is a transmembrane efflux transporter found on the luminal side of the capillary endothelial cells that comprise the blood-brain barrier. This study examined the effect of CJY, an isoflavone, on the levels of a p-glycoprotein substrate, rhodamine123 (Rh123), in rat brain microvessel endothelial cells (RBMEC) and in rat brain homogenate using fluorescence spectrophotometer and high performance liquid chromatography, respectively. In the in vitro study, incubation of RBMEC cells with CJY caused a marked increase in uptake and a notable decrease in efflux of Rh123. The inhibitory effect of the agent on P-gp function was reversible and remained even after the 2.5microM agent was removed from the medium for 120 min. In the in vivo study, CJY also elevated the Rh123 levels in rat brain homogenate after the rats received a 0.45 mg/kg dose of the CJY by slow intravenous injection, indicating that CJY may be a candidate as a p-glycoprotein inhibitor.


Use of soft x-ray diagnostic on the COMPASS tokamak for investigations of sawteeth crash neighborhood and of plasma position using fast inversion methods.
The soft x-ray diagnostic is suitable for monitoring plasma activity in the tokamak core, e.g., sawtooth instability. Moreover, spatially resolved measurements can provide information about plasma position and shape, which can supplement magnetic measurements. In this contribution, fast algorithms with the potential for a real-time use are tested on the data from the COMPASS tokamak. In addition, the soft x-ray data are compared with data from other diagnostics in order to discuss possible connection between sawtooth instability on one side and the transition to higher confinement mode, edge localized modes and productions of runaway electrons on the other side.


Adoptive immunotherapy monitored by micro-MRI in experimental colorectal liver metastasis.
In this study we used the colon carcinoma DHDK12 cell line and generated single metastasis after subcapsular injection in BDIX rats as an experimental tumor model. The aim of the work was to set up in vitro experimental conditions to prepare immune effector cells and in vivo conditions for monitoring the effects of such cells injected as adoptive immunotherapy. Dendritic cells can process tumor cell antigens, induce a T-cell response and be used ex vivo to prepare activated lymphocytes. Lymphocytes were harvested from mesenteric lymph nodes and cocultured with bone marrow-derived autologous dendritic cells previously loaded with irradiated tumor cells. In vitro, the coculture: 1) induced the proliferation of lymphocytes, 2) expanded a preferential subpopulation of T CD8 lymphocytes, and 3) was in favor of lymphocyte cytotoxic activity against the DHDK12 tumor cell line. Activated lymphocytes were injected in the tumor-bearing rat portal vein. Parameters could be set to monitor tumor volume by micro MRI. This monitoring before and after treatment and immunohistochemical examinations revealed that: 1) micro MRI is an appropriate tool to survey metastasis growth in rat, 2) injected lymphocytes increase lesional infiltration with T CD8 cells even 15 days after treatment, 3) a dose of 50 millions lymphocytes is not sufficient to act on the course of the tumor.


Genetic identification of bucktooth parrotfish Sparisoma radians (Valenciennes, 1840) (Labridae, Scarinae) by chromosomal and molecular markers.
Parrotfishes (Labridae, Scarinae) comprise a large marine fish group of difficult identification, particularly during juvenile phase when the typical morphology and coloration of adults are absent. Therefore, the goal of this study was to test cytogenetic markers and DNA barcoding in the identification of bucktooth parrtotfish Sparisoma radians from the northeastern coast of Brazil. Sequencing of cytochrome c oxidase subunit I (COI) confirmed all studied samples as S. radians, and all showed high similarity (99-100%) with Caribbean populations. The karyotype of this species was divergent from most marine Perciformes, being composed of 2n = 46 chromosomes. These consisted of a large number of metacentric and submetacentric pairs with small amounts of heterochromatin and GC-rich single nucleolar organizer regions (NORs) not syntenic to 5S rDNA clusters. These are the first data about DNA barcoding in parrotfish from the Brazilian province and the first refined chromosomal analysis in Scarinae, providing useful data to a reliable genetic identification of S. radians.


Tunable Biogenic Manganese Oxides.
Influence of the conditions for aerobic oxidation of Mn2+(aq) catalysed by the MnxEFG protein complex on the morphology, structure and reactivity of the resulting biogenic manganese oxides (MnOx ) is explored. Physical characterisation of MnOx includes scanning and transmission electron microscopy, and X-ray photoelectron and K-edge Mn, Fe X-ray absorption spectroscopy. This characterisation reveals that the MnOx materials share the structural features of birnessite, yet differ in the degree of structural disorder. Importantly, these biogenic products exhibit strikingly different morphologies that can be easily controlled. Changing the substrate-to-protein ratio produces MnOx either as nm-thin sheets, or rods with diameters below 20 nm, or a combination of the two. Mineralisation in solutions that contain Fe2+(aq) makes solids with significant disorder in the structure, while the presence of Ca2+(aq) facilitates formation of more ordered materials. The (photo)oxidation and (photo)electrocatalytic capacity of the MnOx minerals is examined and correlated with their structural properties.


Remote Clinics and Investigations in Arrhythmia Services: What Have We Learnt During Coronavirus Disease 2019?
The coronavirus disease 2019 (COVID-19) pandemic has had a dramatic impact on the way that medical care is delivered. To minimise hospital attendance by both patients and staff, remote clinics, meetings and investigations have been used. Technologies including hand-held ECG monitoring using smartphones, patch ECG monitoring and sending out conventional Holter monitors have aided remote investigations. Platforms such as Google Meet and Zoom have allowed remote multidisciplinary meetings to be delivered effectively. The use of phone consultations has allowed outpatient care to continue despite the pandemic. The COVID-19 pandemic has resulted in a radical, and probably permanent, change in the way that outpatient care is delivered. Previous experience in remote review and the available technologies for monitoring have allowed the majority of outpatient care to be conducted without obviously compromising quality or safety.


Comparison of two dose levels of stereotactic radiosurgery for 1-3 brain metastases from non-small cell lung cancer.
Two dose groups of patients treated with stereotactic radiosurgery (SRS) alone for 1-3 brain metastases from non-small cell lung cancer (NSCLC) were compared for outcomes. Based on the SRS dose administered to the margins of the brain lesions, 46 patients were assigned to groups treated with 15-18 Gy (n=13) or with 20 Gy (n=33). Seven additional factors were investigated: age (≤ 58 vs. ≥ 59 years), gender, Karnofsky performance score (KPS 70-80 vs. 90-100), number of brain metastases (1 vs. 2-3), histology (adenocarcinoma vs. other) extracerebral metastases and interval from NSCLC diagnosis to SRS (≤ 6 vs. >6 months). Local control rates for 15-18-Gy and 20-Gy groups were 75% and 92% at one year (p=0.043). SRS dose was significant on multivariate analysis (p=0.030). SRS dose was not associated with freedom from new brain metastases (p=0.24) or survival (p=0.37). SRS with 20 Gy resulted in better control of the irradiated metastases than 15-18 Gy did.


Tracking overwintering areas of fish-eating birds to identify mercury exposure.
Migration patterns are believed to greatly influence concentrations of contaminants in birds due to accumulation in spatially and temporally distinct ecosystems. Two species of fish-eating birds, the Double-crested Cormorant (Phalacrocorax auritus) and the Caspian Tern (Hydroprogne caspia) breeding in Lake Ontario were chosen to measure the impact of overwintering location on mercury concentrations ([Hg]). We characterized (1) overwintering areas using stable isotopes of hydrogen (δ(2)H) and band recoveries, and (2) overwintering habitats by combining information from stable isotopes of sulfur (δ(34)S), carbon (δ(13)C), nitrogen (δ(15)N), and δ(2)H in feathers grown during the winter. Overall, overwintering location had a significant effect on [Hg]. Both species showed high [Hg] in (13)C-rich habitats. In situ production of Hg (e.g., through sulfate reducing bacteria in sediments) and allochthonous import could explain high [Hg] in birds visiting (13)C-rich habitats. Higher [Hg] were found in birds with high δ(2)H, suggesting that Hg is more bioavailable in southern overwintering locations. Hotspot maps informed that higher [Hg] in birds were found at the limit of their southeastern overwintering range. Mercury concentrations in winter feathers were positively related to predicted spatial pattern of [Hg] in fish using the National Descriptive Model of Mercury in Fish (NDMMF) based on bird spatial assignment (using δ(2)H). This study indicates that the overwintering location greatly influences [Hg].


TFEB-mediated activation of the lysosome-autophagy system affects the transduction efficiency of adeno-associated virus 2.
Adeno-associated virus (AAV)-mediated gene transfer is an appealing therapeutic option due to AAV's safety profile. Effective delivery of AAV's genetic cargo to the nucleus, however, requires evasion of host cell barriers, including cellular clearance mechanisms mediated by the lysosome-autophagy system. We used AAV serotype 2 to monitor the autophagic response to cellular internalization of AAV and to characterize the effect of AAV-induced activation of autophagy on transgene expression. We found AAV2 internalization to induce activation of transcription factor EB, a master regulator of autophagy and lysosomal biogenesis, and upregulation of the lysosome-autophagy system. We showed that AAV2-induced activation of autophagy parallels a reduction in transgene expression, but also an increase in autophagic clearance of protein aggregates. These results can inform the design of AAV vectors with autophagy-modulating properties for applications ranging from the design of efficient gene delivery vectors to the treatment of diseases characterized by accumulation of autophagic cargo.


Prevalence of and risk factors for adolescent obesity in Southern Appalachia, 2012.
The objective of this study was to examine weight status among southern Appalachian adolescents and to identify risk factors for obesity. We analyzed baseline data from the Team Up for Healthy Living study in 2012. Overall, 19.8% of the sample was overweight, and 26.6% was obese. Boys had higher rates of overweight/obesity than girls (50.5% vs 42.3%). Being male (odds ratio [OR] = 1.79; 95% confidence interval [CI], 1.39-2.29), having a mother with a high school education or less (OR = 1.39; 95% CI, 1.05-1.83), or having a father with a high school education or less (OR = 1.57; 95% CI, 1.17-2.09) was associated with a higher prevalence of obesity and a higher body mass index z score (β = 0.131, 0.160, and 0.043, respectively, P < .05). Parental education could be used to identify adolescents with a higher likelihood of obesity.


Cervicovaginal opsonic activity, immunoglobulins, complement, and soluble fibronectin in healthy women randomly assigned to tampon or napkin use.
The effect of tampon use on cervicovaginal immunoglobulins, complement, fibronectin, and antistaphylococcal opsonic activity was determined in a study of 35 healthy women randomly allocated to either tampon or napkin use for three consecutive menstrual cycles. Sequential cervicovaginal washings were collected on days 3 (+/- 2), 15 (+/- 2), and 25 (+/- 2) of the menstrual cycle before randomization and that 3 months afterward. Prior to randomization, no statistically significant differences were noted for any parameter (with correction for total protein content) between regular tampon users and exclusive napkin users. Following randomization, total opsonic activity during tampon use increased significantly over that in the same women during napkin use (P = .04, Wilcoxon matched-pairs signed-ranks test, two-tailed). This difference was observed only in menstrual samples and not at other sampling times. These data indicate that tampon use has minimal effects on cervicovaginal mucosal immunity and is not associated with aberrations of opsonophagocytosis in the lower female genital tract.


Antioxidative effects of schidandrin B and green tea polyphenols against mercuric chloride-induced hepatotoxicity in rats.
In the present study, we investigated the hepatoprotective effects of schisandrin B (Sch B) and green tea polyphenols (GrTP), two natural antioxidants, against Hg hepatotoxicity. Fifty-six rats were randomly divided into four groups of fourteen animals in each group: control group, Hg-treated group, Sch B pretreated group, and GrTP pretreated group. HgCl2 injection increased Hg accumulation, pathological and ultrastructural injury, lactate dehydrogenase (LDH) level, alanine transaminase (ALT) level, apoptosis, reactive oxygen species (ROS) level, glutathione (GSH) level, and malondialdehyde (MDA) level. HgCl2 injection decreased glutathione peroxidase (GSH-Px) and superoxide dismutase (SOD) activities. Sch B and GrTP protected against Hg-induced dysfunction and liver injury by enhancing the antioxidant defense system. Our results indicate that Sch B and GrTP exert antioxidative effects against Hg-induced hepatotoxicity in rats.


Genotoxic and oxidative stress potential of nanosized and bulk zinc oxide particles in Drosophila melanogaster.
Zinc oxide nanoparticles (ZnONP) are manufactured on a large scale and can be found in a variety of consumer products, such as sunscreens, lotions, paints and food additives. Few studies have been carried out on its genotoxic potential and related mechanisms in whole organisms. In the present study, the in vivo genotoxic activity of ZnONP and its bulk form was assayed using the wing-spot test and comet assay in Drosophila melanogaster Additionally, a lipid peroxidation analysis using the thiobarbituric acid assay was also performed. Results obtained with the wing-spot test showed a lack of genotoxic activity of both ZnO forms. However, when both particle sizes were tested in the comet assay using larvae haemocytes, a significant increase in DNA damage was observed for ZnONP treatments but only at the higher dose applied. In addition, the lipid peroxidation assay showed significant malondialdehyde (MDA) induction for both ZnO forms, but the induction of MDA for ZnONP was higher for the ZnO bulk, suggesting that the observed DNA strand breaks could be induced by mediated oxidative stress. The overall data suggest that the potential genotoxicity of ZnONP in Drosophila can be considered weak according to the lack of mutagenic and recombinogenic effects and the induction of primary DNA damage only at high toxic doses of ZnONP. This study is the first assessing the genotoxic and oxidative stress potential of nano and bulk ZnO particles in Drosophila.


A Photoredox/Sulfide Dual Catalysis System That Uses Sulfide Radical Cations to Promote Alkene Chlorotrifluoromethylation.
Sulfides and their derivatives are among the most important class of reagent in synthetic chemistry. Despite the importance of such compounds, the use of sulfide radical cations in synthetic chemistry is underdeveloped. To address this issue, herein, we describe alkene chlorotrifluoromethylation reactions promoted by photoredox/sulfide dual catalysis systems, which involves sulfide radical cations generated through the oxidation of sulfides by a photoredox catalyst. The high functional group tolerance of this chemistry was demonstrated using natural products and drug molecules as substrate alkenes.


Extramacular Retinal Hole Following Intravitreal Dexamethasone Implant: Case Report
The intravitreal dexamethasone implant Ozurdex is indicated for the treatment of macular edema due to diabetes and branch retinal vein occlusion. While the most common ocular side effects are elevated intraocular pressure and cataract formation, rare complications related to the injection have been reported. We present a case with extramacular retinal hole after Ozurdex injection.


University students' reservations about dietetics as a career choice.
Dietetics students' reservations about their career choice were investigated. In several dietetics programs in various provinces, an in-class or online survey was administered to students in the early or late stages of their education (n=397). Data were coded and analyzed, using descriptive statistics. Chi-square testing for independence was used to establish significant relationships. Forty-three percent (n=149) of those responding to the research question (n=344) indicated they had reservations about a dietetics career, primarily because of internship, salary, and employment concerns. Students enrolled in a coordinated internship/degree program experienced no reservations about internship. Students experienced fewer career reservations when they had made their career decision before grade 12, were influenced by a dietitian, or were in the later stages of their education. Findings have implications for dietetics recruitment, retention, research, and education. An understanding of sources of reservations about career choice will allow policy-makers, researchers, and educators to address issues to ensure that potential professionals are well informed about career components and that educational programs meet students' needs.


Prebiotic effects of a novel combination of galactooligosaccharides and maltodextrins.
Prebiotics are used for stimulating the growth of beneficial microorganisms in the gut. However, it is very difficult to find a suitable prebiotic mixture that exclusively supports the growth of beneficial microbes such as bifidobacteria and lactobacilli. We tested the effects of a prebiotic mixture in vitro by incubating it with fecal samples and in vivo by administration of the prebiotic supplement to healthy adult volunteers, followed by analysis of their fecal microbiota. The effect of the oligosaccharides on bacterial metabolism was studied by analyzing short-chain fatty acid (SCFA) production in vitro and the SCFA pattern for the stool samples of volunteers. In the in vitro test, a higher proportion of bifidobacteria (25.77%) was seen in the total bacterial population after cultivation on a prebiotic mixture than on the control medium (7.94%). The gram-negative anaerobe count significantly decreased from 8.70 to 6.40 log CFU/g (from 35.21% to 0.60%) and the Escherichia coli count decreased from 7.41 to 6.27 log CFU/g (from 1.78% to 0.44%). Administration of a prebiotic mixture in vivo (9 g of galactooligosaccharides [GOS]+1 g of maltodextrins; daily for 5 days) significantly increased the fecal bifidobacterial count from 9.45 to 9.83 log CFU/g (from 40.80% to 53.85% of total bacteria) and reduced the E. coli count from 7.23 to 6.28 log CFU/g (from 55.35% to 45.06% of total bacteria). The mixture comprising GOS and maltodextrins thus exhibited bifidogenic properties, promoting the performance of bifidobacteria by boosting their growth and inhibiting the growth of undesirable bacteria.


Functional regulation of platelet/endothelial cell adhesion molecule-1 by TGF-beta 1 in promonocytic U-937 cells.
Platelet endothelial cell adhesion molecule-1 (PECAM-1) is a widely distributed cell adhesion molecule present on monocytes, macrophages, and monocytic cell lines. Treatment of the promonocytic cell line U-937 with TGF-beta 1 induces homotypic cellular aggregations, simultaneous with an increase in surface expression and specific transcripts of PECAM-1. The TGF-beta-induced cell adhesion phenomena are not dependent on LFA-1, intercellular adhesion molecule-1 (ICAM-1), very late Ag-4 (VLA-4), or very late Ag-5 (VLA-5). However, the phenomena seem to be directly mediated by PECAM-1 because 1) it is inhibited by the addition of Abs or an antisense oligonucleotide specific for PECAM-1; and 2) TGF-beta 1-treated U-937 cells bind to PECAM-1-expressing mouse transfectant fibroblasts, but not to mock transfectants. In addition, this aggregation phenomena are divalent cation-dependent and requires the integrity of the cytoskeleton. Analysis of the intracellular signaling pathways indicates that TGF-beta 1 induces protein kinase C activity, as well as PECAM-1 phosphorylation and association with cytoskeletal components. Furthermore, in this model, an autocrine mechanism for releasing the bioactive form of TGF-beta 1 operates, allowing PECAM-1 activation. These results provide evidence that TGF-beta 1 regulates PECAM-1 function by increasing the expression and activating the adhesion of PECAM-1 in monocytic cells. These two mechanisms seem to be necessary for adhesion because independent inhibition of either expression or activation of PECAM-1 leads to abrogation of cellular aggregation.


Polyunsaturated fatty acid (PUFA) changes in serum and liver of undernourished rats given dietary vitamin B6 supplementation.
The influence of vitamin B6 on linoleic (LA), alpha-linolenic (ALA), arachidonic (AA), eicosapentaenoic (EPA) and docosahexaenoic (DHA) acid content in serum and liver of rats fed with protein-energy deficient diets for 90 d, was studied. To estimate the effect of dietary supplementation with vitamin B6 on the composition and level of fatty acids in the serum and liver of rats, two experiments were performed. In these experiments control rats were fed ad libitum semisynthetic isocaloric diets of 1,466.5 kJ/100 g (350 kcal/100 g) throughout 90 d while the examined rats were offered 50% and 30% of the previously determined daily intake of the diet consumed in the control group. The experimental diet was supplemented with vitamin B6 to the level 4-times higher than in the control diet. A reduction to the half consumption of a standard diet supplemented with vitamin B6 caused a significant decrease of LA and ALA in blood serum at 30 and 60 d. At 90 d of the experiment the value of LA was lower and the content of AA was higher in comparison to the control group. After 30 d of consumption of vitamin B6 enriched diet in rats subjected to feed restriction to only 30% of the control intake, an increase of ALA and a decrease of AA, EPA and DHA were noticed in serum. At 60 d an increase of DHA was observed. Ninety days of feeding this diet caused a significant increase of AA level. Feeding animals for 90 d with a vitamin B6 enriched diet, with limited consumption to 50%, caused a significant increase of ALA content in liver. Further limitation of this diet consumption to 30%, caused a significant decrease of LA and ALA and an increase of EPA content.


Supplementation of Syzygium cumini seed powder prevented obesity, glucose intolerance, hyperlipidemia and oxidative stress in high carbohydrate high fat diet induced obese rats.
Obesity and related complications have now became epidemic both in developed and developing countries. Cafeteria type diet mainly composed of high fat high carbohydrate components which plays a significant role in the development of obesity and metabolic syndrome. This study investigated the effect of Syzygium cumini seed powder on fat accumulation and dyslipidemia in high carbohydrate high fat diet (HCHF) induced obese rats. Male Wistar rats were fed with HCHF diet ad libitum, and the rats on HCHF diet were supplemented with Syzygium cumini seed powder for 56 days (2.5% w/w of diet). Oral glucose tolerance test, lipid parameters, liver marker enzymes (AST, ALT and ALP) and lipid peroxidation products were analyzed at the end of 56 days. Moreover, antioxidant enzyme activities were also measured in all groups of rats. Supplementation with Syzygium cumini seed powder significantly reduced body weight gain, white adipose tissue (WAT) weights, blood glucose, serum insulin, and plasma lipids such as total cholesterol, triglyceride, LDL and HDL concentration. Syzygium cumini seed powder supplementation in HCHF rats improved serum aspartate amino transferase (AST), alanine amino transferase (ALT), and alkaline phosphatase (ALP) activities. Syzygium cumini seed powder supplementation also reduced the hepatic thiobarbituric acid reactive substances (TBARS) and elevated the antioxidant enzyme superoxide dismutase (SOD) and catalase (CAT) activities as well as increased glutathione (GSH) concentration. In addition, histological assessment showed that Syzygium cumini seed powder supplementation prevented inflammatory cell infiltration; fatty droplet deposition and fibrosis in liver of HCHFD fed rats. Our investigation suggests that Syzygium cumini seed powder supplementation prevents oxidative stress and showed anti-inflammatory and antifibrotic activity in liver of HCHF diet fed rats. In addition, Syzygium cumini seed powder may be beneficial in ameliorating insulin resistance and dyslipidemia probably by increasing lipid metabolism in liver of HCHF diet fed rats.


Repair of deoxyribonucleic acid in ultraviolet light-sensitive and -resistant Dictyostelium discoideum strains.
Some responses of the cellular slime mold Dictyostelium discoideum to ultraviolet light (UV) irradiation were investigated by analyzing two aspects of deoxyribonucleic acid (DNA) excision repair in the vegetative cells: (i) the fate of thymine-containing dimers and (ii) the production and rejoining of single-strand breaks. Experiments were done with the parental, radiation-resistant NC-4 strain and with the radiation-sensitive gammas-13 strain. The majority (greater than 85%) of the thymine-containing dimers produced in both strains by an energy fluence of 100/Jm2 were removed from the acid-insoluble DNA fraction within the first 3 to 4 h of reincubation in the dark. Moreover, as measured by alkaline sucrose gradients, single-strand breaks appeared in the DNA of both NC-4 and gammas-13 irradiated cells very rapidly and at low temperatures. This was presumed to be a result of the incision (nicking) step of excision repair as performed by UV-specific endonuclease(s). In NC-4 the time required for dimer excision correlated with the sealing of breaks as well as with the UV-induced division delays. In gammas-13 the single-strand breaks were closed at a slower rate than in NC-4. However, this was not accompanied by more extensive division delays.


Adenocarcinoma of the endometrium with psammoma bodies.
Two patients with endometrial carcinoma containing psammoma bodies are presented. The necessity of distinguishing this neoplasm from an ovarian carcinoma is stressed. The rarity of this finding and its histogenesis are discussed, and the literature on the subject is reviewed.


Negative correlation between ICA persistence and beta cell restoration after IDDM diagnosis.
We have studied 27 insulin-dependent diabetic patients since diagnosis for a period of six months; at diabetes onset and later on, ICA were found in 17 of them, whereas they were undetectable in 10 patients: age was remarkably homogeneous in the two groups. At diabetes onset, no significant differences were found in insulin requirement between ICA positive and ICA negative patients; however, six months after diagnosis, we observed that insulin requirement to keep metabolic control was significantly higher in ICA positive than in ICA negative subjects (0.515 +/- 0.2 U/Kg versus 0.22 +/- 0.15 U/Kg, p less than 0.001). Only one remission has been detected in ICA positive group (insulin requirement less than 0.25 U/Kg), while four ICA negative patients had complete remission and three had partial remission. ICA positive patients showed fasting C-peptide values higher than ICA negative (0.5 +/- 0.28 ng/ml versus 1.4 +/- 0.5 ng/m.; p less than 0.001, at rest; 1.1 +/- 0.6 ng/ml; versus 2.6 +/- 1.0 ng/ml, 6 minutes after stimulus; p less than 0.05). Our study suggests that presence and persistence of ICA may be considered an early and predictive marker for a worse beta cell function restoration resulting in a higher insulin requirement.


Source identification of PM2.5 in Steubenville, Ohio using a hybrid method for highly time-resolved data.
A new source-type identification method, Reduction and Species Clustering Using Episodes (ReSCUE), was developed to exploit the temporal synchronicity typically observed between ambient species in high time resolution fine particulate matter (PM2.5) data to form clusters that vary together. High time-resolution (30 min) PM2.5 sampling was conducted for a month during the summer of 2006 in Steubenville, OH, an EPA designated nonattainment area for the U.S. National Ambient Air Quality Standards (NAAQS). When the data were evaluated, the species clusters from ReSCUE matched extremely well with the source types identified by EPA Unmix demonstrating that ReSCUE is a valuable tool in identifying source types. Results from EPA Unmix show that contributions to PM2.5 are mostly from iron/steel manufacturing (36% ± 9%), crustal matter (33% ± 11%), and coal combustion (11% ± 19%). More importantly, ReSCUE was useful in (i) providing objective data driven guidance for the number of source factors and key fitting species for EPA Unmix, and (ii) detecting tenuous associations between some species and source types in the results derived by EPA Unmix.


Small Biopsies Misclassify up to 35% of PD-L1 Assessments in Advanced Lung Non-Small Cell Lung Carcinomas.
Pembrolizumab is an FDA-approved immune-checkpoint (IC) inhibitor that targets programmed cell death protein PD-1, and recent phase III trials have demonstrated its superiority over chemotherapy in the treatment of patients with advanced non-small cell lung cancer (NSCLC). Eligibility for treatment with Pembrolizumab is based on demonstration of PD-L1 expression on tumoral cells using the approved companion test 22C3 PharmDx (Dako). Access to the drug depends on a tumor proportion score (TPS) expressing the PD-L1 protein above predetermined cutoffs. The scoring interpretation guide requires a minimum of 100 viable cells to be considered adequate for evaluation. Recent studies have questioned the adequacy of the sampling process when small biopsies are utilized. To further explore this concern, the viable tumor area of 426 consecutive NSCLC biopsies and surgical excisions submitted for PD-L1 assessment was measured and recorded with corresponding PD-L1 expression. About 14.6% of all biopsies measured <2 mm creating 2 groups (<2 mm and ≥2 mm) whose PD-L1 categories distribution [negative (<1%), low expressor (≥1% and <50%), and positive (≥50%)] were compared. Results were significantly different between both groups (χ test; P=0.0012). To help understand this difference, 1,407,000 in silico simulated biopsies of various sizes were performed on 201 numerical tumors created from digitalized full sections and analyzed. Not only the same results shown in actual biopsies were reproduced, but the model calculated that up to 35% of very small biopsies were misclassified including a mixture of false negative and false positive results. The percentage decreased to 10% with a threshold of 5 mm. In era of precision medicine, appropriate sampling is more than ever critical to achieve accurate assessment of the NSCLC PD-L1. Ignored in most clinical trials, recording of biopsy size would permit refining data analysis and increase predictive accuracy of current and future biomarkers.


[Endoscopic ligation of maxillary and sphenopalatine artery for intractable epistaxis].
We report the endoscopic ligation of the maxillary and sphenopalatine arteries for the treatment of intractable epistaxis. From March 2003 to February 2005, 17 patients (12 men, 5 women) with epistaxis were hospitalized in our department. Patient age ranged from 25 to 83 years, with an average age of 62 years. 8 patients were successfully treated using the conventional packing method, 2 patients were treated using electrocauterization, and 1 patient with macroglobulinemia was treated using plasma exchange therapy. 6 patients underwent endoscopic ligation of the maxillary and sphenopalatine arteries while under general anesthesia. The post operative courses were uneventful, and no recurrent bleeding has been noted. Endscopic ligation of the maxillary and sphenopalatine arteries is safer than intraarterial embolization and less invasive than conventional surgical approach for the ligation of maxillary artery. This technique appears to be a safe and effective surgical treatment for patients with intractable epistaxis.


Perspectives of Frequent Users of Emergency Departments on a Case Management Intervention: A Qualitative Study.
Effective management of frequent users of emergency departments (FUED) remains challenging. Case management (CM) has shown to improve patient quality of life while reducing ED visits and associated costs. However, little data is available on FUED's perception of CM outside of North America to further improve CM implementation. Explore the FUED's perspectives about CM in Switzerland. Semi-structured qualitative interviews eliciting FUED's experiences of CM were conducted among 20 participants (75% female; mean age = 40.6, SD = 12.8) across 6 hospital ED. Inductive content analysis. Most participants were satisfied with the CM program. In particular, FUEDs identified the working relationship with the case manager (cm) as key for positive outcomes, and also valued the holistic evaluation of their needs and resources. Overall, patients reported increased motivation and health literacy, as well as facilitated interactions within the healthcare system. Conversely, a small number of participants reported negative views on CM (ie, stigmatization, lack of concrete outcomes). Barriers identified were cm's lack of time, COVID-19's negative impact on CM organization, as well as lack of clarity on the objectives of CM. FUED perceived CM as useful, in particular establishing a working relationship with the cm. Our results suggest that CM can be further improved by (1) professionals remaining non-judgmental toward FUED, (2) making sure the aims and objectives of the CM are understood by the participants, and (3) allowing more time for the cm to carry out their work.


[Osteopetrosis with carbonic anhydrase II deficiency: report of 24 cases].
Twenty four patients suffering from osteopetrosis caused by carbonic anhydrase II deficiency are colliged. This pathology seems to be frequent in Tunisia. Mental retardation is present in 52%, 85% of patients have short stature and 25% have optic atrophy. All affected subjects show craniofacial disproportion and dental anomalies. Twenty patients have at least one bone fracture. Metabolic acidosis is constant: it is profound during the first life decade. A severe selective reduction of carbonic anhydrase II in erythrocyte is confirmed in 18 cases. Osteosclerosis and defective skeletal modelling are constant, cerebral calcification can be seen at the scanner approximately at the age of two years and six months. All patients are homozygous for a splice junction mutation in intron 2 of the carbonic anhydrase II gene, this mutation does not seem to protect patients from bone fractures nor induce a severe metabolic acidosis.


Screening for renal disease: the role of the practice nurse.
Screening tests for renal disease are inexpensive, easy to carry out and simple to interpret. Nurses in general practice can provide screening for high-risk populations and interpret the results of these tests. Practice nurses can also provide patient information about renal disease, which will both improve patients' quality of life and save time and money for the NHS.


Identification of differentially expressed genes and enriched pathways in lung cancer using bioinformatics analysis.
Lung cancer is the leading cause of cancer‑associated mortality worldwide. The aim of the present study was to identify the differentially expressed genes (DEGs) and enriched pathways in lung cancer by bioinformatics analysis, and to provide potential targets for diagnosis and treatment. Valid microarray data of 31 pairs of lung cancer tissues and matched normal samples (GSE19804) were obtained from the Gene Expression Omnibus database. Significance analysis of the gene expression profile was used to identify DEGs between cancer tissues and normal tissues, and a total of 1,970 DEGs, which were significantly enriched in biological processes, were screened. Through the Gene Ontology function and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis, 77 KEGG pathways associated with lung cancer were identified, among which the Toll‑like receptor pathway was observed to be important. Protein‑protein interaction network analysis extracted 1,770 nodes and 10,667 edges, and identified 10 genes with key roles in lung cancer with highest degrees, hub centrality and betweenness. Additionally, the module analysis of protein‑protein interactions revealed that 'chemokine signaling pathway', 'cell cycle' and 'pathways in cancer' had a close association with lung cancer. In conclusion, the identified DEGs, particularly the hub genes, strengthen the understanding of the development and progression of lung cancer, and certain genes (including advanced glycosylation end‑product specific receptor and epidermal growth factor receptor) may be used as candidate target molecules to diagnose, monitor and treat lung cancer.


Regulation of carbohydrate and fatty acid utilization by L-carnitine during cardiac development and hypoxia.
This study is designed to investigate whether substrate preference in the myocardium during the neonatal period and hypoxia-induced stress is controlled intracellularly or by extracellular substrate availability. To determine this, the effect of exogenous L-carnitine on the regulation of carbohydrate and fatty acid metabolism was determined during cardiac stress (hypoxia) and during the postnatal period. The effect of L-carnitine on long chain (palmitate) and medium chain (octonoate) fatty acid oxidation was studied in cardiac myocytes isolated from less than 24 h old (new born; NB), 2 week old (2 week) and hypoxic 4 week old (HY) piglets. Palmitate oxidation was severely decreased in NB cells compared to those from 2 week animals (0.456+/-0.04 vs. 1.207+/-0.52 nmol/mg protein/30 min); surprisingly, cells from even older hypoxic animals appeared shifted toward the new born state (0.695+/-0.038 nmol/mg protein/30 min). Addition of L-carnitine to the incubation medium, which stimulates carnitine palmitoyl-transferase I (CPTI) accelerated palmitate oxidation 3 fold in NB and approximately 2 fold in HY and 2 week cells. In contrast, octanoate oxidation which was greater in new born myocytes than in 2 week cells, was decreased by L-carnitine suggesting a compensatory response. Furthermore, oxidation of carbohydrates (glucose, pyruvate, and lactate) was greatly increased in new born myocytes compared to 2 week and HY cells and was accompanied by a parallel increase in pyruvate dehydrogenase (PDH) activity. The concentration of malonyl-CoA, a potent inhibitor of CPTI was significantly higher in new born heart than at 2 weeks. These metabolic data taken together suggest that intracellular metabolic signals interact to shift from carbohydrate to fatty acid utilization during development of the myocardium. The decreased oxidation of palmitate in NB hearts probably reflects decreased intracellular L-carnitine and increased malonyl-CoA concentrations. Interestingly, these data further suggest that the cells remain compliant so that under stressful conditions, such as hypoxia, they can revert toward the neonatal state of increased glucose utilization.


The effect of changing from whole-cell to acellular pertussis vaccine on the epidemiology of hospitalized children with pertussis in Canada.
Between July 1997 and April 1998, universal childhood immunization programs in Canada changed from using a whole-cell pertussis to a 5-component acellular pertussis-containing vaccine. To assess effects on pertussis epidemiology of this nationwide change, we analyzed hospitalizations during 1991-2004 using the Canadian Immunization Monitoring Program, Active (IMPACT) pertussis database. IMPACT is an active surveillance network based in 12 pediatric tertiary-care hospitals across Canada. Characteristics of hospitalized cases of pertussis were compared by type of vaccine received or by birth date (if immunization records were unavailable or the child was unvaccinated). Age-stratified incidence rates were calculated by year and vaccine type. Two thousand ninety-six cases of pertussis were admitted to IMPACT centers, 1174 during the whole-cell vaccine program (WCV-P) and 842 during the acellular vaccine program (ACV-P). Pertussis incidence among children <5 years old decreased significantly during the ACV-P, causing an increase in the residual proportion of cases either too young to be immunized (<2 months old: ACV-P 39% versus WCV-P 26.1%; P < 0.0001) or too young for a second dose (2-3 months old: 42.9% versus 34.2%, respectively; P < 0.0001). A significantly smaller proportion of cases (ACV-P 15.1% versus WCV-P 27.3%) occurred in infants who were old enough (4-11 months of age) to have received 2 or 3 doses of vaccine. With ACV-P, pertussis hospitalizations in children 4-59 months old decreased in frequency, consistent with improved vaccine effectiveness, but remained prominent among very young infants. Improved control strategies are needed to reduce infections among infants too young for pertussis vaccination.


Metabolic Regulation of the Epitranscriptome.
An emergent theme in cancer biology is that dysregulated energy metabolism may directly influence oncogenic gene expression. This is due to the fact that many enzymes involved in gene regulation use cofactors derived from primary metabolism, including acetyl-CoA, S-adenosylmethionine, and 2-ketoglutarate. While this phenomenon was first studied through the prism of histone and DNA modifications (the epigenome), recent work indicates metabolism can also impact gene regulation by disrupting the balance of RNA post-transcriptional modifications (the epitranscriptome). Here we review recent studies that explore how metabolic regulation of writers and erasers of the epitranscriptome (FTO, TET2, NAT10, MTO1, and METTL16) helps shape gene expression through three distinct mechanisms: cofactor inhibition, cofactor depletion, and writer localization. Our brief survey underscores similarities and differences between the metabolic regulation of the epigenome and epitranscriptome, and highlights fertile ground for future investigation.


Respiratory cryptosporidiosis in HIV-seronegative children in Uganda: potential for respiratory transmission.
Respiratory cryptosporidiosis is recognized as a late-stage complication in persons with human immunodeficiency virus (HIV) infection and AIDS. However, respiratory signs and symptoms are common in otherwise healthy children with intestinal cryptosporidiosis, which suggests that respiratory infection may occur in immunocompetent hosts. We recruited children 9-36 months of age who presented with diarrhea to Mulago Hospital in Kampala, Uganda, from November 2007 through January 2009. Children with stool samples positive or negative for Cryptosporidium species were selected for further evaluation, including sputum induction in those with cough or unexplained respiratory signs and collection of saliva and blood specimens. Sputum samples were subjected to comprehensive bacteriologic testing, and both sputum and saliva specimens were tested for Cryptosporidium species by nested polymerase chain reaction. Of 926 fecal samples screened, 116 (12.5%) were positive for Cryptosporidium. Seventeen (35.4%) of 48 sputum samples tested from children with positive stool samples were positive for Cryptosporidium. Sixteen (94.1%) of the 17 children with confirmed respiratory cryptosporidiosis were HIV seronegative, and 10 (58.8%) of 17 children were not malnourished. None of the 12 sputum specimens from children with negative stool samples tested positive for Cryptosporidium (P = .013, compared with children who tested positive for Cryptosporidium in the stool). Parasite DNA was detected in only 2 (1.9%) of 103 saliva samples (P < .001, compared with sputum samples). Respiratory cryptosporidiosis was documented in one-third of HIV-seronegative children who were tested. These novel findings suggest the potential for respiratory transmission of cryptosporidiosis. Trial registration. ClinicalTrials.gov identifier: NCT00507871.


Intrathecal therapy with ziconotide: clinical experience and considerations on its use.
Ziconotide is a synthetic peptide equivalent of an w-conotoxin, obtained from the marine snail Conus magus, which acts by blocking N-type calcium channels in the spinal cord, reducing the perception of pain. It is a newly marketed drug, exclusively for intrathecal use, indicated for severe chronic pain. Ziconotide came to the physicians' table with both doubts and promises; to determine its safety and efficacy, one of the largest and most well-designed randomized double-blind studies in the history of intrathecal therapy was undertaken, and this drug demonstrated efficacy in relieving chronic pain in patients coming from many years of different therapies and therapy failures. However, the pain relief came with some adverse effects, which are few compared with morphine's adverse effects but in some cases could undermine the course of therapy with this conotoxin. The experience described in this paper began in June 2007 and gave us the opportunity to analyze how the conotoxin works outside of the papers. We noted differences between the well-known activity of morphine on pain and mood, and the more focused action of ziconotide on pain. In addition, it is important to consider the lack of addiction, opioid-induced hyperalgesia and other systemic effects that are common with morphine. These are the reasons why the Polyanalgesic Consensus Conference of 2007 put ziconotide in the first line of intrathecal therapy management.


Measurement of antibodies against glutamic acid decarboxylase 65 (GADA): two new 125I assays compared with [35S]GAD 65-ligand binding assay.
Recently, 65-kDa glutamic acid decarboxylase (GAD 65) antibodies (GADA) have been introduced as autoimmune markers in blood to confirm the diagnosis of insulin-dependent diabetes mellitus (IDDM). In this study, to evaluate two new assays that use 125I-labeled GAD 65, we assayed samples from 100 children with recent onset of diabetes and 100 control children, the results were compared with those of a [35S]GADA assay and with results for islet cell antibodies (ICA), the conventional autoimmune marker. Receiver operating characteristic (ROC) curve analysis showed one of the new assays (from RSR) to be more sensitive (P = 0.01) than the comparison ([35S]GADA) assay, whereas the second new assay (from Elias) was less sensitive (P < 0.001). The GADA frequency at 97.5% specificity was greatest in the comparison assay: 63 of 100 vs 41 of 100 (P < 0.01) and 53 of 100 (P = 0.16) in the RSR and Elias assays, respectively. Almost all GADA-positive patients had ICA, but one-third of the ICA-positive patients was GADA-negative. Accordingly, adding GADA analysis results to ICA testing increased the frequency of detection of autoimmune markers only slightly (from 81% to 85%). In conclusion, at 97.5% specificity the [35S]GADA assay seemed to be more efficient than the 125I assays, although the difference was significant only for the Elias 125I assay. Antigen-specific antibodies other than GADA may explain the difference in GADA and ICA frequencies.


Effects of Sho-Saiko-to on hepatocarcinogenesis and 8-hydroxy-2'-deoxyguanosine formation.
Oxidative stress plays an important role in hepatocarcinogenesis. Although Sho-saiko-to (TJ-9), a Japanese herbal medicine which has been recently administered to patients with chronic liver disease in Japan, prevents hepatocarcinogenesis, the mechanism by which TJ-9 protects against cancer development is not fully understood. 8-Hydroxy-2'-deoxyguanosine (8-OHdG), a DNA adduct by reactive oxygen species, is known as a parameter of genetic risk for hepatocarcinogenesis. To clarify whether the preventive effect on hepatocarcinogenesis by TJ-9 is dependent on 8-OHdG, the effect on 8-OHdG levels by TJ-9 was examined by using high-performance liquid chromatography-mass spectrometry (LC-MS) in a diethylnitrosamine (DEN)-induced hepatocarcinogenesis model of male Fisher rats. TJ-9 reduced the number of preneoplastic cells, detected as the glutathione S transferase P (GST-P)-positive hepatocytes, and inhibited the development of liver tumors. TJ-9 also significantly decreased the formation of 8-OHdG, as indicated by LC-MS and immunohistochemical analysis. In addition, ornithine decarboxylase (ODC) activity and the number of proliferating cell nuclear antigen (PCNA)-positive cells were not altered. An electron paramagnetic resonance spin-trapping technique showed that TJ-9 scavenges hydroxyl radicals in a dose-dependent manner. In conclusion, the results of the present study suggest that TJ-9 prevents hepatocarcinogenesis in association with inhibition of 8-OHdG formation.


Evaluation of the Effect of Passive Smoking on Lactoferrin and AST on 12 - 15 Years Old Children and Adolescents.
Passive smokers are involuntarily exposed to cigarette or tobacco smoke and as known, inhalation of environmental tobacco smoke is a serious threat. There is little information about the effect of passive smoking on salivary markers and periodontal indices. This study investigated the effect of passive smoking on lactoferrin and AST in 12 - 15 years old children and adolescents. This case-control analytic correlation type study with no-convenience random sampling method was performed on 160 children aged 12 - 15 who had smokers in their families. The eligible children were divided into two equal groups; 80 cot(+) children as case group and 80 cot(-) children as control group, matched according to age, sex and plaque index. Plaque index was obtained from all subjects. 2 cc unstimulated salivary samples were collected by spitting method. The collected specimens were tested by lactoferrin and AST kits in biochemistry were measured on the day of sampling laboratory. Gingival index Loe and Silness (GI) and Probing Pocket Depth (PPD). Mean and Standard Deviation of PPD and GI was 2.01 ± 0.077 and 1.53 ± 0.055 in experimental group and 1.93 ± 0.073 and 1.49 ± 0.046 in control group respectively (P < 0.001). The Mean and Standard Deviation parameters of lactoferrin and AST, in the experimental group was 38.66 ± 25.15 and 13.45 ± 6.33 and in the control group 10.18 ± 6.82 and 6.53 ± 2.65 group, respectively (P < 0.001). Passive smoking can be effective on inflammatory process of periodontal and salivary biomarkers related to inflammation. Lactoferrin was 11 - 104 in case group and 0.5 - 38 in control group. Aspartat aminotransferase in case group was 2.64 - 30.43 and in control group it was 2.16 - 12.02.


Life-management strategies of selection, optimization, and compensation: measurement by self-report and construct validity.
The authors examined the usefulness of a self-report measure for elective selection, loss-based selection. optimization, and compensation (SOC) as strategies of life management. The expected 4-factor solution was obtained in 2 independent samples (N = 218, 14-87 years; N = 181, 18-89 years) exhibiting high retest stability across 4 weeks (r(tt) = .74-82). As expected, middle-aged adults showed higher endorsement of SOC than younger and older adults. Moreover, SOC showed meaningful convergent and divergent associations to other psychological constructs (e.g., thinking styles, NEO) and evinced positive correlations with measures of well-being which were maintained after other personality and motivational constructs were controlled for. Initial evidence on behavioral associations involving SOC obtained in other studies is summarized.


Exploring Dynamic Risk Prediction for Dialysis Patients.
Despite substantial advances in the treatment of end-stage renal disease, mortality of hemodialysis patients remains high. Several models exist that predict mortality for this population and identify patients at risk. However, they mostly focus on patients at a particular stage of dialysis treatment, such as start of dialysis, and only use the most recent patient data. Generalization of such models for predictions in later periods can be challenging since disease characteristics change over time and the evolution of biomarkers is not adequately incorporated. In this research, we explore dynamic methods which allow updates of initial predictions when patients progress in time and new data is observed. We compare a Dynamic Bayesian Network (DBN) to regularized logistic regression models and a Cox model with landmarking. Our preliminary results indicate that the DBN achieves satisfactory performance for short term prediction horizons, but needs further refinement and parameter tuning for longer horizons.


miR-134 in extracellular vesicles reduces triple-negative breast cancer aggression and increases drug sensitivity.
Exosomes (EVs) have relevance in cell-to-cell communication carrying pro-tumorigenic factors that participate in oncogenesis and drug resistance and are proposed to have potential as self-delivery systems. Advancing on our studies of EVs in triple-negative breast cancer, here we more comprehensively analysed isogenic cell line variants and their EV populations, tissues cell line variants and their EV populations, as well as breast tumour and normal tissues. Profiling 384 miRNAs showed EV miRNA content to be highly representative of their cells of origin. miRNAs most substantially down-regulated in aggressive cells and their EVs originated from 14q32. Analysis of miR-134, the most substantially down-regulated miRNA, supported its clinical relevance in breast tumours compared to matched normal breast tissue. Functional studies indicated that miR-134 controls STAT5B which, in turn, controls Hsp90. miR-134 delivered by direct transfection into Hs578Ts(i)8 cells (in which it was greatly down-regulated) reduced STAT5B, Hsp90, and Bcl-2 levels, reduced cellular proliferation, and enhanced cisplatin-induced apoptosis. Delivery via miR-134-enriched EVs also reduced STAT5B and Hsp90, reduced cellular migration and invasion, and enhanced sensitivity to anti-Hsp90 drugs. While the differing effects achieved by transfection or EV delivery are likely to be, at least partly, due to specific amounts of miR-134 delivered by these routes, these EV-based studies identified miRNA-134 as a potential biomarker and therapeutic for breast cancer.


Evaluation of MLC leaf positioning using a scanning liquid ionization chamber EPID.
A method was developed to determine the accuracy of multileaf collimator (MLC) positioning using transmitted dose maps measured by a scanning liquid ionization chamber electronic portal imaging device (SLIC-EPID). Several MLC fields were designed, using the Varian C-series standard MLC-80, as reference fields for open fields. The MLC leaves were then shifted from the reference positions along the direction of MLC leaf movement towards the central axis from 0.1 to 1.6 mm. The electronic portal images (EPIs), acquired for each case, were converted to two-dimensional dose maps using an appropriate calibration method and the relative dose difference maps were then calculated. The experiment was then performed at non-zero gantry angles in the presence of an anthropomorphic phantom for typical prostate and head and neck fields. Several standard edge detection algorithms were also used in order to find the shifted MLC leaf position. In addition, the short-term reproducibility of MLC leaf positioning was evaluated using the above-mentioned methods. It was found that the relationship between the relative dose difference and MLC leaf spatial displacement is linear. A variation of 0.2 mm in leaf position leads to approximately 4% change in the relative dose values for open fields. The variation of the relative dose difference for phantom studies depends on the phantom positioning and the EPI normalization. From the standard edge detection algorithms, used in the current study, the 'Canny' algorithm was found to be the optimum method to identify the minimum detectable MLC leaf displacements with a precision of approximately 0.1 mm for all cases. However, the result of edge detection algorithms generally is binary and there is no additional information compared to the relative dose maps. The reproducibility of MLC positions was found to be within 0.3 mm. In conclusion, a SLIC-EPID can be used for regular quality assurance (QA) of MLC leaf positioning. Despite significant difference in the pixel size of the acquired SLIC-EPIs, it can be concluded that the SLIC-EPID can be used for MLC quality assurance protocols with similar accuracy compared to amorphous silicon (a-Si) EPID results.


Environmental variability, reliability of information and the timing of migration.
The timing of migration and migratory steps is highly relevant for fitness. Because environmental conditions vary between years, the optimal time for migration varies accordingly. Therefore, migratory animals could clearly benefit from acquiring information as to when it is the best time to migrate in a specific year. Thus, environmental predictability and variability are fundamental characteristics of migration systems but their relationship and consequence for migratory progression has remained unexplored. We develop a simple dynamic model to identify the optimal migration behaviour in environments that differ in predictability, variability and the number of intermediate stop-over sites. Our results indicate that higher predictability along migration routes enables organisms to better time migration when phenology deviates from its long-term average and thus, increases fitness. Information is particularly valuable in highly variable environments and in the final migration-step, i.e. before the destination. Furthermore, we show that a general strategy for obtaining information in relatively uninformative but variable environments is using intermediate stop-over sites that enable migrants to better predict conditions ahead. Our study contributes to a better understanding of the relationship between animal movement and environmental predictability-an important, yet underappreciated factor that strongly influences migratory progression.


Severe post-influenza (H1N1) encephalitis involving pulvinar nuclei in an adult patient.
Neurological complications of H1N1 infections are mostly found in children, but rare cases of acute encephalopathy and post-infectious encephalitis such as acute disseminated encephalomyelitis (ADEM) have been described in adults. We report a case of an adult presenting with a progressive and severe encephalopathy that developed after H1N1 respiratory infection resolution. Cerebrospinal fluid (CSF) analysis was normal, including negative PCR for herpes simplex virus, H1N1, influenza B and JC virus, and absent oligoclonal IgG bands in CSF and serum. Initial CT scan was normal, but later MRI showed posterior multifocal leucoencephalopathy with pulvinar sign. The delayed neurological findings together with the ancillary investigation, namely the MRI pattern with both grey and white matter involvement, raised the possibility of a post-infectious process, rather than an acute encephalitis. Despite aggressive immunotherapy, the patient experienced severe neurological sequelae. Early recognition of ADEM manifestations by those dealing with H1N1 infection is important as early immunotherapy may improve the prognosis.


Update in Veterinary Radiation Oncology: Focus on Stereotactic Radiation Therapy.
Stereotactic radiotherapy (SRT) involves the precise delivery of highly conformal, dose-intense radiation to well-demarcated tumors. Special equipment and expertise are needed, and a unique biological mechanism distinguishes SRT from other forms of external beam radiotherapy. Families find the convenient schedules and minimal acute toxicity of SRT appealing. Common indications in veterinary oncology include nasal, brain, and bone tumors. Many other solid tumors can also be treated, including spinal, oral, lung, heart-base, liver, adrenal, and prostatic malignancies. Accessibility of SRT is improving, and new data are constantly emerging to define parameters for appropriate case selection, radiation dose prescription, and long-term follow-up."


Oesophageal tuberculosis.
The case discussed is that of a previously healthy 48-year-old female who presented with a week long history of epigastric pain and continuing weight loss. A series of investigations and supporting literature alluded to a diagnosis of oesophageal tuberculosis (TB), and antituberculous medication was commenced accordingly. An accompanying discussion considers the incidence, differential diagnoses, pathogenesis, clinical features, investigations and aspects of management of oesophageal TB.


Enkephalin is essential for the molecular and behavioral expression of cocaine sensitization.
Behavioral sensitization to cocaine is associated to neuroadaptations that contribute to addiction. Enkephalin is highly expressed in mesocorticolimbic areas associated with cocaine-induced sensitization; however, their influence on cocaine-dependent behavioral and neuronal plasticity has not been explained. In this study, we employed a knockout (KO) model to investigate the contribution of enkephalin in cocaine-induced behavioral sensitization. Wild-type (WT) and proenkephalin KO mice were treated with cocaine once daily for 9 days to induce sensitization. Additionally, to clarify the observations in KO mice, the same procedure was applied in C57BL/6 mice, except that naloxone was administered before each cocaine injection. All animals received a cocaine challenge on days 15 and 21 of the treatment to evaluate the expression of locomotor sensitization. On day 21, microdialysis measures of accumbal extracellular dopamine, Western blotting for GluR1 AMPA receptor (AMPAR), phosphorylated ERK2 (pERK2), CREB (pCREB), TrKB (pTrkB) were performed in brain areas relevant for sensitization from KO and WT and/or naloxone- and vehicle pre-treated animals. We found that KO mice do not develop sensitization to the stimulating properties of cocaine on locomotor activity and on dopamine release in the nucleus accumbens (NAc). Furthermore, pivotal neuroadaptations such as the increase in pTrkB receptor, pERK/CREB and AMPAR related to sensitized responses were absent in the NAc from KO mice. Consistently, full abrogation of cocaine-induced behavioral and neuronal plasticity after naloxone pre-treatment was observed. We show for first time that the proenkephalin system is essential in regulating long-lasting pivotal neuroadaptations in the NAc underlying behavioral sensitization to cocaine.


Participation of the C-terminal region of the D1-polypeptide in the first steps in the assembly of the Mn4Ca cluster of photosystem II.
Amino acid residue D1-Asp(170) of the D1-polypeptide of photosystem II was previously shown to be implicated in the binding and oxidation of the first manganese to be assembled into the Mn(4)Ca cluster of the oxygen-evolving complex (OEC). According to recent x-ray crystallographic structures of photosystem II, D1-Glu(333) is proposed to participate with D1-Asp(170) in the coordination of Mn4 of the OEC. Other residues in the C-terminal region of the D1-polypeptide are proposed to coordinate nearby manganese of the cluster. Site-directed replacements in Synechocystis sp. PCC 6803 at D1-His(332), D1-Glu(333), D1-Asp(342), D1-Ala(344), and D1-Ser(345) were examined with regard to their ability to influence the binding and oxidation of the first manganese in manganese-depleted photosystem II core complexes. Direct and indirect measurements reveal in all mutants, but most marked in D1-Glu(333) replaced by His, an impaired ability of Mn(2+) to reduce Y(Z)., indicating a reduced ability (elevated K(m)) compared with WT to bind and oxidize the first manganese of the OEC. The effect on the K(m) of these mutations is, however, considerably weaker than some of those constructed at D1-Asp(170) (replacement by Asn, Ala, and Ser). These observations imply that the C-terminal residues ultimately involved in manganese coordination contribute to the high affinity binding at D1-Asp(170) likely through electrostatic interactions. That these residues are far from D1-Asp(170) in the primary structure of the D1-polypeptide, imply that the C terminus of the D1-polypeptide is already close to its mature conformation at the first stages of assembly of the Mn(4)Ca cluster.


Lipid management for cardiovascular risk reduction in type 1 diabetes.
To review the recent evidence for lipid management in type 1 diabetes (T1D) for cardiovascular risk reduction. Individuals with T1D are at increased risk for cardiovascular morbidity and mortality, with atherosclerosis beginning as early as adolescence. Elevated low-density lipoprotein cholesterol (LDL-C), triglycerides, and lipoprotein (a) are associated with increased cardiovascular risk in T1D. Although high-density lipoprotein cholesterol (HDL-C) in T1D is often normal or higher than in nondiabetic controls, HDL in T1D has structural alterations, which make it proatherogenic rather than cardioprotective. Similarly, although LDL-C is not particularly elevated in T1D, LDL still contributes to cardiovascular risk. Studies in individuals with diabetes have primarily included T2D participants, with a much smaller number of T1D participants; such studies have shown that lipid-lowering therapies, such as statins, ezetimibe, and proprotein convertase subtilisin/kexin type 9 (PCSK9) inhibitors reduce LDL-C levels and cardiovascular events in both those with and without diabetes. Individuals with T1D have increased cholesterol absorption, suggesting that ezetimibe may be particularly effective in T1D. Results of the REDUCE-IT trial show cardiovascular risk reduction from high-dose omega-3 fatty acid (Icosapent Ethyl) therapy in patients with diabetes (primarily type 2 diabetes), independent of triglyceride lowering, but similar data in T1D are currently lacking. Individuals with T1D are at high risk of cardiovascular disease, necessitating close lipid monitoring and management from adolescence through adulthood.


Distribution and activity of alternatively spliced Alzheimer amyloid peptide precursor and scrapie PrP mRNAs on rat brain polysomes.
Mammalian brains contain low levels of the Alzheimer amyloid precursor variants (AAPPs) and the normal form of the scrapie agent protease-resistant protein (PrPc); however, their mRNAs are readily detectable. To understand these discrepancies we have investigated some aspects of the translational regulation of these mRNAs. An accurate blot-hybridization procedure was developed to measure absolute amounts of mRNA. Rat brain contains the following mRNA levels (ng/g tissue) AAPP(695), 170; AAPP(751/770), 63; PrPc, 144; actin, 615; glyceraldehyde-3-phosphate dehydrogenase (G3PDH), 359; ferritin, 148. The method was also used to determine the distribution of mRNAs between translationally active polysomes and translationally inactive ribonucleoprotein protein particles (mRNPs). More than 90% of G3PDH and actin mRNAs were associated with polysomal RNA; whereas, ferritin light chain mRNA was predominantly (90%) in mRNP RNA. The degree of cross-contamination of mRNPs with polysomes was less than 10%. Probes specific for the scrapie PrP protein and the AAPP(695) splice junction revealed that 70% of these mRNAs were associated with polysomes. One-half of AAPP(751/770) mRNAs (which comprise 20-30% of all AAPP mRNA in brain) were found in polysomes. We conclude therefore that both scrapie and AAPP mRNAs are subject to translational regulation in rat brain. Evidence from in vitro translational experiments confirm the message distribution determined by blot hybridization and corroborate the hypothesis that AAPP is subject to partial post-transcriptional regulation. Nevertheless, the low tissue levels of AAPP and PrPc must result primarily from their relatively rapid turnover.


Seasonal changes of plasma membrane H(+)-ATPase and endogenous ion current during cambial growth in poplar plants.
The plasma membrane H(+)-ATPase (PM H(+)-ATPase), potassium ions, and endogenous ion currents might play a fundamental role in the physiology of cambial growth. Seasonal changes of these parameters were studied in twigs of Populus nigra and Populus trichocarpa. Monoclonal and polyclonal antibodies against the PM H(+)-ATPase, x-ray analysis for K(+) localization and a vibrating electrode for measurement of endogenous ion currents were used as probes. In dormant plants during autumn and winter, only a slight immunoreactivity against the PM H(+)-ATPase was found in cross sections and tissue homogenates, K(+) was distributed evenly, and the density of endogenous current was low. In spring during cambial growth, strong immunoreactivity against a PM H(+)-ATPase was observed in cambial cells and expanding xylem cells using the monoclonal antibody 46 E5 B11 F6 for fluorescence microscopy and transmission electron microscopy. At the same time, K(+) accumulated in cells of the cambial region, and strong endogenous current was measured in the cambial and immature xylem zone. Addition of auxin to dormant twigs induced the formation of this PM H(+)-ATPase in the dormant cambial region within a few days and an increase in density of endogenous current in shoot cuttings within a few hours. The increase in PM H(+)-ATPase abundance and in current density by auxin indicates that auxin mediates a rise in number and activity of an H(+)-ATPase in the plasma membrane of cambial cells and their derivatives. This PM H(+)-ATPase generates the necessary H(+)-gradient (proton-motive force) for the uptake of K(+) and nutrients into cambial and expanding xylem cells.


An inversion technique for the calculation of embedding potentials.
A new embedding method to include local correlation in large systems is proposed. In this method the density of the whole system, calculated via density functional theory approaches, is partitioned in two pieces, one corresponding to the subsystem of interest and the rest to the environment. In the second step, an embedding potential is obtained iteratively using as a driving force the self-repulsion due to the density difference, in a similar form as proposed by Zhao et al. [Phys. Rev. A 50, 2138 (1994)], to obtain the "exact" exchange-correlation functional. Such potential is added to the Fock equation to build the localized molecular orbitals which are further used to include the local electronic correlation in the subsystem of interest. This method is an alternative to the previous DFT-based embedding methods first proposed by Wesolowski and Washell [J. Phys. Chem. 97, 8050 (1993)] and after enhanced by Govind et al. [J. Chem. Phys. 110, 7677 (1999)] and adapted to metal extended systems, which use density functionals to describe the kinetic energy contribution to the embedding potential, whose precise form has been largely treated in the literature and its crucial role is discussed here. The method is applied to hydrogen chains and its van der Waals interaction with H(2). The results obtained are in very good agreement with exact calculations performed on the whole system, which demonstrates that the method proposed is a very promising route to introduce correlation in large systems.


Foetal experience: lifelong consequences.
Modification of the foetal environment has lifelong consequences for endocrine function and behaviour. Maternal stress through activation of the hypothalamus-pituitary-adrenal axis can lead to increased anxiety levels and stress reactivity in offspring. Furthermore, these effects can pass across generations. The race is now on to understand the mechanisms involved and to find ways to prevent or reverse the effects of an "adverse" early environment in human populations.


Critique of the analysis of UpToDate.com on the treatment of painful vertebral compression fractures: time to update UpToDate.
The treatment of painful vertebral compression fractures has changed substantially since the introduction of vertebroplasty in the mid-1980s and balloon kyphoplasty in the late 1990s. Both procedures were widely accepted with the vertebral fractures treated reaching 150,000 per annum in 2009 prior to the publication of 2 randomized controlled trials comparing vertebroplasty with a sham treatment published in the New England Journal of Medicine in August 2009. Since then, there has been a flood of information on vertebral augmentation and balloon kyphoplasty. It is worth evaluating this information especially because it relates to current recommendations that are often followed blindly by medical and administrative groups unfamiliar with either the procedure or the high level of evidence surrounding vertebral augmentation. To streamline the evaluation of some current recommendations, we limited the analysis to the recommendations found on UpToDate.com. This Web site is an evidence-based, peer-reviewed source of information available for patients, doctors, health insurance companies, and population-based medical decision-making.