Humoral immune response in dogs and cats vaccinated against rabies in southeastern Brazil.
Brazil holds annual nationwide public campaigns to vaccinate dogs and cats against rabies. The presence of rabies antibodies in these animals, which are among the main transmitters of rabies to humans, is a good indicator that they are immunized and protected. In the present study we analyzed 834 serum samples from dogs and cats from the Southeast of Brazil (Presidente Prudente and Dracena cities), 12 months after the 2009 vaccination campaign. We used the technique known as rapid fluorescent focus inhibition test (RFFIT) and considered reactant those sera with values higher 0.5 IU/mL. Reactant sample results in Presidente Prudente were 153 (51.0%) for dogs and 59 (32.6%) for cats, and in Dracena 110 (52.1%) for dogs and 71 (50.0%) for cats. We discussed vaccine coverage of animals involved in this experiment, and observed low titers < 0.5 IU/mL, especially in cats from Presidente Prudente. According to the results presented in our experiment, we suggest that titers below 0.5 IU/mL are worrisome and that, for multiple reasons, animals should be immunized against rabies in the period between public vaccination campaigns. Hence, the desired vaccine coverage was not accomplished, especially among cats from Presidente Prudente.


A One-Pot, One-Operation
Enantiomerically pure 2,1-benzothiazines are readily prepared by a one-pot, one-operation procedure consisting of a palladium-catalyzed aryl amination of an ortho-bromobenzaldehyde with a sulfoximine followed by an intramolecular condensation reaction [Eq. (1)]. BINAP=1,1'-binaphthalene-2,2'-diylbis(diphenylphosphane).


Association of 5-HTT gene polymorphism, platelet MAO activity, and drive for thinness in a population-based sample of adolescent girls.
Several lines of evidence suggest that alterations in serotonergic activity contribute to the pathophysiology of abnormal eating behaviors. Since platelet monoamine oxidase (MAO) activity and the 5-HT transporter gene promoter polymorphism (5-HTTLPR) have been associated with eating disorders, the knowledge from a population-based sample may provide useful information which changes in 5-HT function observed in eating disorders represent trait vs. state effects. The sample was based on both cohorts of the Estonian Children Personality, Behavior and Health Study (ECPBHS). The current study was conducted during the second follow-up where altogether 82% from the original sample was recruited. EDI-2 subscales--Drive for Thinness and Bulimia--were used to determine eating attitudes and behaviors. Platelet MAO activity was measured and the participants were genotyped for the 5-HTTLPR. Allelic variation of 5-HTTLPR or platelet MAO activity were not independently associated with drive for thinness or binge eating, but girls homozygous for the 5-HTTLPR long allele and with high platelet MAO activity, both considered indicators of a higher capacity 5-HT system, exhibited higher scores of drive for thinness. The results suggest that drive for thinness is the highest in girls with the presence of two markers of higher serotonergic capacity.


Sarm1 activation produces cADPR to increase intra-axonal Ca++ and promote axon degeneration in PIPN.
Cancer patients frequently develop chemotherapy-induced peripheral neuropathy (CIPN), a painful and long-lasting disorder with profound somatosensory deficits. There are no effective therapies to prevent or treat this disorder. Pathologically, CIPN is characterized by a "dying-back" axonopathy that begins at intra-epidermal nerve terminals of sensory neurons and progresses in a retrograde fashion. Calcium dysregulation constitutes a critical event in CIPN, but it is not known how chemotherapies such as paclitaxel alter intra-axonal calcium and cause degeneration. Here, we demonstrate that paclitaxel triggers Sarm1-dependent cADPR production in distal axons, promoting intra-axonal calcium flux from both intracellular and extracellular calcium stores. Genetic or pharmacologic antagonists of cADPR signaling prevent paclitaxel-induced axon degeneration and allodynia symptoms, without mitigating the anti-neoplastic efficacy of paclitaxel. Our data demonstrate that cADPR is a calcium-modulating factor that promotes paclitaxel-induced axon degeneration and suggest that targeting cADPR signaling provides a potential therapeutic approach for treating paclitaxel-induced peripheral neuropathy (PIPN).


Rapid degradation of cyclic peroxides by titanium and antimony chlorides.
First responders face extraordinary risks when dealing with organic peroxides such as TATP due to the extreme sensitivity of this class of explosives, and to a lack of a robust chemical means of safe and rapid neutralisation. The Lewis acids TiCl4 and SbCl3 have been found to demonstrate a novel degradation mechanism, with TiCl4 degrading a model cyclic peroxide in minutes when used in a two-fold excess, or ∼3 hours at half equivalence. The products cannot re-form peroxide compounds as is the case with acid degradation, suggesting the two mechanisms are fundamentally different. The Lewis acids mediate a rearrangement reaction in the cyclic peroxide backbone leading to relatively innocuous products through deactivation of oxidising O. Sub-stoichiometric TiCl4 reactions highlight a secondary reaction pathway that also leads to some oxidative chlorination products, possibly mediated by an unconfirmed titanium-oxychloride species. SbCl3 was found to exhibit similar reactivity to TiCl4, although at a slower rate. A mechanism is proposed, consistent with the observations for both stoichiometric and sub-stoichiometric quantities of TiCl4.


From diagnosis to prognosis: A paradigm shift for multimodal imaging in assessing diabetic macular edema.
Diabetic macular edema (DME) is one of the leading causes of visual impairment in patients with diabetes. Multimodal imaging (MMI) has allowed a shift from DME diagnosis to prognosis. Although there are no accepted guidelines, MMI may also lead to treatment customization. Several study groups have tried to identify structural biomarkers that can predict treatment response and long-term visual prognosis. The purpose of this editorial is to review currently proposed optical coherence tomography (OCT) and optical coherence tomography angiography (OCT-A) biomarkers.


Computerized tomographic morphometric analysis of the cervical spine.
Detailed knowledge of cervical canal and transverse foramens' morphometry is critical for understanding the pathology of certain diseases and for proper preoperative planning. Lateral x-rays do not provide the necessary accuracy. A retrospective morphometric study of the cervical canal was performed at the authors' institution to measure mean dimensions of sagittal canal diameter (SCD), right and left transverse foramens' sagittal (SFD) and transverse (TFD) diameters and minimum distance between spinal canal and transverse foramens (dSC-TF) for each level of the cervical spine from C1-C7, using computerized tomographic scans, in 100 patients from the archives of the Emergency Room. Significant differences for SCD were detected between C1 and the other levels of the cervical spine for both male and female patients. For the transverse foramen, significant differences in sagittal diameters were detected at C3, C4, C5 levels. For transverse diameters, significant differences at C3 and C4 levels. A significant difference of the distance between the transverse spinal foramen and the cervical canal was measured between left and right side at the level of C3. This difference was equally observed to male and female subjects. CT scan can replace older conventional radiography techniques by providing more accurate measurements on anatomical elements of the cervical spine that could facilitate diagnosis and preoperative planning, thus avoiding possible trauma to the vertebral arteries during tissue dissection and instrument application.


Expectations of post-partum care among pregnant women living in the north of Sweden.
To describe expectations of post-partum care among pregnant women living in the north of Sweden and whether personality determines preference for care systems. The time for post-partum care on maternity wards has been reduced in Western countries. This, along with the reduction in special medical treatments offered and the closure of small hospitals has affected pregnant women and their families. Data was extracted from a questionnaire and a personality instrument (SSP) that were completed during November 2002. In the northernmost county of Sweden, 140 pregnant women completed the questionnaire; of these, 120 completed the SSP instrument. Of the women who participated, 61.3% wanted to be discharged 72 hours after childbirth, irrespective of the distance between the hospital and home. To have access to maternity ward staff and the decision to be discharged were described as being the most important issues in maternity ward care. The infant's father was expected to be the most important person in the post-partum period. Women ranked the opportunity to decide for themselves when to be discharged from the maternity ward as important, which can be interpreted as a strong signal that the women want to be in control of the care they receive. Midwives have to focus more on the woman and her family's individual needs, and to include the father as a person who also needs support and to provide resources for him.


Hydrogenase genes from Rhizobium leguminosarum bv. viciae are controlled by the nitrogen fixation regulatory protein nifA.
Rhizobium leguminosarum bv. viciae expresses an uptake hydrogenase in symbiosis with peas (Pisum sativum) but, unlike all other characterized hydrogen-oxidizing bacteria, cannot express it in free-living conditions. The hydrogenase-specific transcriptional activator gene hoxA described in other species was shown to have been inactivated in R. leguminosarum by accumulation of frameshift and deletion mutations. Symbiotic transcription of hydrogenase structural genes hupSL originates from a -24/-12 type promoter (hupSp). A regulatory region located in the -173 to -88 region was essential for promoter activity in R. leguminosarum. Activation of hupSp was observed in Klebsiella pneumoniae and Escherichia coli cells expressing the K. pneumoniae nitrogen fixation regulator NifA, and in E. coli cells expressing R. meliloti NifA. This activation required direct interaction of NifA with the essential -173 to -88 regulatory region. However, no sequences resembling known NifA-binding sites were found in or around this region. NifA-dependent activation was also observed in R. etli bean bacteroids. NifA-dependent hupSp activity in heterologous hosts was also absolutely dependent on the RpoN sigma-factor and on integration host factor. Proteins immunologically related to integration host factor were identified in R. leguminosarum. The data suggest that hupSp is structurally and functionally similar to nitrogen fixation promoters. The requirement to coordinate nitrogenase-dependent H2 production and H2 oxidation in nodules might be the reason for the loss of HoxA in R. leguminosarum and the concomitant NifA control of hup gene expression. This evolutionary acquired control would ensure regulated synthesis of uptake hydrogenase in the most common H2-rich environment for rhizobia, the legume nodule.


Basic and clinical aspects of copper.
An oxygen-rich atmosphere obligated living organisms to cope with reactive oxygen species (O2-, H2O2, OH*) that were the unavoidable by-products of cellular metabolism. As a redox cofactor Cu was selected as a co-catalyst for numerous biological processes, many involving the utilization of oxygen. Inadequate or excessive intake of Cu can be pathogenic and life-threatening. Mutations to genes that code for Cu-transporting ATPase enzymes are the molecular basis of Wilson and Menkes diseases and more recently Cu has been identified as a preemptory factor in amyloid and prion diseases. This review is dedicated to bringing historical and timely information on Cu transport, metabolism and homeostasis to the attention of those not familiar with this important mineral. Other comprehensive reviews are available to the interested readers.


Heparin Binding to an Engineered Virus-like Nanoparticle Antagonist.
The anticoagulant activity of heparin administered during medical interventions must be reversed to restore normal clotting, typically by titrating with protamine. Given the acute toxicity associated with protamine, we endeavored to generate safer heparin antagonists by engineering bacteriophage Qβ virus-like particles (VLPs) to display motifs that bind heparin. A particle bearing a single amino acid change from wild-type (T18R) was identified as a promising candidate for heparin antagonism. Surface potential maps generated through molecular modeling reveal that the T18R mutation adds synergistically to adjacent positive charges on the particle surface, resulting in a large solvent-accessible cationic region that is replicated 180 times over the capsid. Chromatography using a heparin-sepharose column confirmed a strong interaction between heparin and the T18R particle. Binding studies using fluorescein-labeled heparin (HepFL) resulted in a concentration-dependent change in fluorescence intensity, which could be perturbed by the addition of unlabeled heparin. Analysis of the fluorescence data yielded a dissociation constant of approximately 1 nM and a 1:1 binding stoichiometry for HepFL:VLP. Dynamic light scattering (DLS) experiments suggested that T18R forms discrete complexes with heparin when the VLP:heparin molar ratios are equivalent, and in vitro clotting assays confirmed the 1:1 binding stoichiometry as full antagonism of heparin is achieved. Biolayer interferometry and backscattering interferometry corroborated the strong interaction of T18R with heparin, yielding Kd ∼ 1-10 nM. These biophysical measurements further validated T18R, and VLPs in general, for potential clinical use as effective, nontoxic heparin antagonists.


Decongestion improves cell-mediated immunity in postmastectomy arm lymphoedema: a pilot study.
Chronic lymphoedematous limbs have an increased propensity for infections and primary or secondary malignant tumours. It has been attributed to suppressed delayed-type hypersensitivity measured in lymphoedemas related to Stewart-Treves syndrome, Kaposi's sarcoma or breast cancer treatment. Cell-mediated immunity is an effective defence mechanism against bacteria, fungi, viruses and tumour cells. We aimed to examine whether decongestive lymphoedema therapy could improve cell-mediated immunity in breast cancer treatment-related lymphoedema (BCRL). Eight women with unilateral BCRL were included in this study. At baseline, tuberculin skin test (TST) was performed on the volar surfaces of the forearms of the affected and non-affected sides using 0.5, 1 and 5 tuberculin units in the form of three consecutive injections with 3-cm spaces in-between, and arm volumes were measured using the Kuhnke's disc model. Decongestive lymphatic therapy was given to swollen arms in 10 consecutive working days. At the end of intensive decongestion, TST on affected side and bilateral volumetry were repeated. Baseline test using undiluted (5 units) and fivefold diluted (1 unit) tuberculin solutions has shown significant differences (P < 0.05) between the mean sizes (11.81 ± 2.32 and 7.75 ± 1.92; 7.12 ± 1.12 and 5.12 ± 0.91 respectively) in favour to healthy arms. Post therapeutically, the mean sizes were significantly increased (P < 0.05) in the dilutions of 1 : 1 and 1 : 5 (7.75 ± 1.92 and 10.56 ± 1.23 mm, 5.12 ± 0.91 and 5.93 ± 1.74 mm respectively). Significant increase in TST sizes suggests that decongestive lymphatic therapy is able to partially restore impaired cellular immune function in BCRL.


SCN5A mutations and polymorphisms in patients with ventricular fibrillation during acute myocardial infarction.
Mutations in the SCN5A gene encoding the Nav1.5 channel α-subunit are known to be risk factors of arrhythmia, including Brugada Syndrome and Long QT syndrome subtype 3. The present study focused on the role of SCN5A variants in the development of ventricular fibrillation (VF) during acute myocardial infarction (AMI). Since VF during AMI is the major cause of sudden death in the Western world, SCN5A mutations represent genetic risk factors for sudden death. By exon re-sequencing, the entire coding region and flanking intron regions were sequenced in 46 AMI/VF+ patients. In total, nine single nucleotide variants were identified of which four represented common single nucleotide polymorphisms (SNPs; 87G>A, 1673A>G, IVS16‑6C>T and 5457T>A). Only five rare variants were identified, each in only one patient. Only two of the rare variants represented missense mutations (3578G>A and 4786T>A). The common SNPs and the missense mutations were also genotyped using polymerase chain reaction methods in 79 AMI/VF‑ patients and 480 healthy controls. The SNPs did not demonstrate significant differences in allele and genotype frequencies between the study groups. The 3578G>A mutation was identified in one out of the 480 controls, whereas the 4786T>A mutation was not present in AMI/VF- patients and controls. In conclusion, the majority of AMI/VF+ patients demonstrated a wild type sequence or common SNPs in SCN5A. Only two out of 46 (4.3%) AMI/VF+ patients revealed mutations that may be involved in Nav1.5 dysfunction and VF. However, this requires further functional validation.


Patellar skin surface temperature by thermography reflects knee osteoarthritis severity.
Digital infrared thermal imaging is a means of measuring the heat radiated from the skin surface. Our goal was to develop and assess the reproducibility of serial infrared measurements of the knee and to assess the association of knee temperature by region of interest with radiographic severity of knee Osteoarthritis (rOA). A total of 30 women (15 Cases with symptomatic knee OA and 15 age-matched Controls without knee pain or knee OA) participated in this study. Infrared imaging was performed with a Meditherm Med2000™ Pro infrared camera. The reproducibility of infrared imaging of the knee was evaluated through determination of intraclass correlation coefficients (ICCs) for temperature measurements from two images performed 6 months apart in Controls whose knee status was not expected to change. The average cutaneous temperature for each of five knee regions of interest was extracted using WinTes software. Knee x-rays were scored for severity of rOA based on the global Kellgren-Lawrence grading scale. The knee infrared thermal imaging procedure used here demonstrated long-term reproducibility with high ICCs (0.50-0.72 for the various regions of interest) in Controls. Cutaneous temperature of the patella (knee cap) yielded a significant correlation with severity of knee rOA (R = 0.594, P = 0.02). The skin temperature of the patellar region correlated with x-ray severity of knee OA. This method of infrared knee imaging is reliable and as an objective measure of a sign of inflammation, temperature, indicates an interrelationship of inflammation and structural knee rOA damage.


Allergy and the cardiovascular system.
The most dangerous and life-threatening manifestation of allergic diseases is anaphylaxis, a condition in which the cardiovascular system is responsible for the majority of clinical symptoms and for potentially fatal outcome. The heart is both a source and a target of chemical mediators released during allergic reactions. Mast cells are abundant in the human heart, where they are located predominantly around the adventitia of large coronary arteries and in close contact with the small intramural vessels. Cardiac mast cells can be activated by a variety of stimuli including allergens, complement factors, general anesthetics and muscle relaxants. Mediators released from immunologically activated human heart mast cells strongly influence ventricular function, cardiac rhythm and coronary artery tone. Histamine, cysteinyl leukotrienes and platelet-activating factor (PAF) exert negative inotropic effects and induce myocardial depression that contribute significantly to the pathogenesis of anaphylactic shock. Moreover, cardiac mast cells release chymase and renin that activates the angiotensin system locally, which further induces arteriolar vasoconstriction. The number and density of cardiac mast cells is increased in patients with ischaemic heart disease and dilated cardiomyopathies. This observation may help explain why these conditions are major risk factors for fatal anaphylaxis. A better understanding of the mechanisms involved in cardiac mast cell activation may lead to an improvement in prevention and treatment of systemic anaphylaxis.


Electroconvulsive shock therapy and maximum binding of platelet tritiated imipramine binding in depression.
We studied the tritiated imipramine binding values in platelets from 12 hospitalized untreated patients with endogenous depression and found a significant decrease in maximum binding (Bmax) values compared with a control population of the same age and sex. There were no changes in the equilibrium dissociation affinity constant values between the untreated depressives and the control population. After at least six sessions of electroconvulsive therapy and at the time when a significant clinical improvement of depression was confirmed, the Bmax value of tritiated imipramine binding in platelets was slightly increased but was still significantly below that of the control values. However, when six of these patients were reexamined after 12 to 18 months, at a time when they were euthymic, the Bmax of tritiated imipramine binding in platelets was found in the same range as the values of the control population. Our results indicate that clinical improvement precedes the changes in Bmax of tritiated imipramine binding in platelets from depressed patients. The tritiated imipramine binding in platelets is a useful biologic marker in affective disorders. Furthermore, our results suggest that tritiated imipramine binding in platelets may be a state-dependent biologic marker in depression.


Stonefish envenomations of the hand--a local marine hazard: a series of 8 cases and review of the literature.
The Singapore shoreline has changed significantly in recent years, but the stonefish (Synanceia sp.) continues to inhabit our native waters and human envenomation still occurs regularly. In this paper, we document their clinical course, review our management experience and propose a treatment algorithm. Envenomation is associated with appreciable local morbidity, excruciating pain and gross oedema of the affected limb. Severe systemic morbidity and deaths have been reported in the literature but are very rare. Prompt first aid with immersion in hot water (45 degrees C) inactivates the venom. Supplementary analgesia, tetanus prophylaxis and broad-spectrum antibiotics are recommended. Specific antivenom is available and indicated for severe envenomations with systemic symptoms. Complicated puncture wounds and retained spines require surgical debridement. Eight cases of stonefish envenomations to the hand were treated over the last 1.25 years (October 2001 to January 2003). Length of hospital stay averaged 3.9 days. There were no deaths or significant systemic morbidity, but 1 case required surgical debridement for local necrosis. Complete resolution of swelling, with return to full function, occurred on average by 8.2 days. Prompt recognition of envenomation, early first aid and hot water soaks result in rapid relief of pain and symptoms. Our local experience suggests that the majority of stonefish envenomations do not result in significant or protracted morbidity and require only supportive management. Systemic morbidity and mortalities are rare.


Visualization of sodium dynamics in the kidney by magnetic resonance imaging in a multi-site study.
Sodium magnetic resonance imaging (MRI) is a powerful, non-invasive technique to assess sodium distribution within the kidney. Here we undertook pre-clinical and clinical studies to quantify the corticomedullary sodium gradient in healthy individuals and in a porcine model of diuresis. The results demonstrated that sodium MRI could detect spatial differences in sodium biodistribution across the kidney. The sodium gradient of the kidney changed significantly after diuresis in the pig model and was independent of blood electrolyte measurements. Thus, rapid sodium MRI can be used to dynamically quantify sodium biodistribution in the porcine and human kidney.


Bedroom media: One risk factor for development.
Mass media have numerous effects on children, ranging from influencing school performance to increased or reduced aggression. What we do not know, however, is how media availability in the bedroom moderates these effects. Although several researchers have suggested that bedroom media may influence outcomes by displacing other activities (the displacement hypothesis) or by changing the content of media consumed (the content hypothesis), these have rarely been tested directly. This study tested both hypotheses using several outcomes that are associated with bedroom media and some of the underlying mediating mechanisms. The hypotheses were tested using 3 longitudinal samples of varying methods, age, duration, and country. The results indicate that children who have bedroom media are likely to watch larger amounts of screen time which displaced important activities, such as reading and sleeping, which mediated later negative outcomes such as poor school performance. Bedroom media also influence risk for obesity and video game addiction. Children with bedroom media are also likely to be exposed to more media violence. The violent content increased normative beliefs about aggression, which increased physical aggression, providing support for the content hypothesis. This study demonstrates that media can have effects not just from what they show, but also because of what children are not exposed to. Bedroom media are therefore a robust risk factor for several aspects of child development. (PsycINFO Database Record


Recombinant human ulinastatin improves immune dysfunction of dendritic cells in septic mice by inhibiting endoplasmic reticulum stress-related apoptosis.
Urinary trypsin inhibitor (UTI), also known as ulinastatin, has been reported to protect multiple organs against inflammation- and/or injury-induced dysfunction. In the present study, we aimed to investigate the immunomodulation effects of a recombinant human ulinastatin (urinary trypsin inhibitor, UTI) (rhUTI) on splenic dendritic cells (DCs) in cecal ligation and puncture (CLP)-induced septic mice. CLP mice were treated with rhUTI intramuscularly at 0, 12, and 24 h after procedure. Splenic CD11c+ DCs were isolated and accessed with flow cytometry for apoptotic or phenotypic analysis. Protein markers and cytokines were determined with Western blotting or ELISA. Treatment with rhUTI could markedly upregulate levels of costimulatory molecules (CD80, CD86) and MHC-II on surface of the splenic DC in CLP mice. The apoptotic rate of splenic DCs was decreased in CLP mice after rhUTI treatment. The survival rate of septic mice was increased after treatment with rhUTI. In addition, protein level of markers in endoplasmic reticulum stress (ERS)-related apoptotic pathways (including GRP78, caspase-12, and CHOP) were obviously down-regulated in the rhUTI-treated group when compared with the CLP group. These results indicate that rhUTI protects CLP-induced sepsis in mice by improving immune response of splenic DCs and inhibiting the excessive ERS-mediated apoptosis.


Classification of Actigraphy Records from Bipolar Disorder Patients Using Slope Entropy: A Feasibility Study.
Bipolar Disorder (BD) is an illness with high prevalence and a huge social and economic impact. It is recurrent, with a long-term evolution in most cases. Early treatment and continuous monitoring have proven to be very effective in mitigating the causes and consequences of BD. However, no tools are currently available for a massive and semi-automatic BD patient monitoring and control. Taking advantage of recent technological developments in the field of wearables, this paper studies the feasibility of a BD episodes classification analysis while using entropy measures, an approach successfully applied in a myriad of other physiological frameworks. This is a very difficult task, since actigraphy records are highly non-stationary and corrupted with artifacts (no activity). The method devised uses a preprocessing stage to extract epochs of activity, and then applies a quantification measure, Slope Entropy, recently proposed, which outperforms the most common entropy measures used in biomedical time series. The results confirm the feasibility of the approach proposed, since the three states that are involved in BD, depression, mania, and remission, can be significantly distinguished.


Concentration of phosphoribosyl pyrophosphate in renal hypertrophy. Contrasting effects of early diabetes and unilateral nephrectomy.
Studies were made of the renal phosphoribosyl pyrophosphate (PPRibP) content and PPRibP synthetase (EC 22.214.171.124) activity in rats diabetic for 5, 14 or 20 days, or unilaterally nephrectomized (UN) for 5 days, and in doubly lesioned animals. Approximately equal degrees of renal enlargement were found after 5 days diabetes or 5 days UN. In the doubly lesioned animals the increment of growth was additive. Unilateral nephrectomy of 5 days duration, in contrast with diabetes, had no effect on the PPRibP content of the contralateral kidney, nor did it modify the renal PPRibP content when performed on animals diabetic for 5, 14 or 20 days. The activity of PPRibP synthetase was unaffected by diabetes, UN or diabetes +UN. The results pinpoint a stage of nucleotide synthesis which is differentially affected by the two stimuli, in line with evidence for differences in regulation of nucleic acid turnover in the two conditions.


Causal inference in studies of preterm babies: a simulation study.
Using a simple simulation, we illustrate why associations estimated from studies restricted to preterm births cannot be interpreted causally. Data simulation involving a hypothetical cohort of fetuses who may be healthy or have one or more of four pathological factors (termed A through D, increasing in severity) with known effects on gestational length and risk of mortality. We focus on babies born at ≤32 weeks of gestation. We visually represent the simulated population and compare the association between A (which may represent pre-eclampsia) and neonatal death. We then repeat the exercise with D (standing in for chorioamnionitis) as the exposure of interest. Odds ratios of neonatal death in the simulated data. In most weeks, and for both A and D, the calculated odds ratios are substantially biased and underestimate the true risk of neonatal death associated with each pathology. For example, factor A has a true causal odds ratio of 1.50, yet it appears protective among births ≤32 weeks (estimated crude odds ratio 0.39; gestational age-adjusted odds ratio 0.71). Among very preterm births, virtually all babies are born with pathologies that increase the risk of adverse outcomes. Hence, babies exposed to one factor (e.g. pre-eclampsia) are compared with babies who have a mix of other pathologies. Such selection bias affects studies carried out among very preterm births (e.g. where pre-eclampsia appears to reduce risk of adverse neonatal outcomes). Selection bias affects studies of preterm births, complicating interpretation.


D-alanyl-lipoteichoic acid in Lactobacillus casei: secretion of vesicles in response to benzylpenicillin.
Vesicles containing lipoteichoic acid (LTA) have been isolated from Lactobacillus casei ATCC 7469 grown in the presence of either benzylpenicillin or D-cycloserine. These cell wall antibiotics enhanced the rate of LTA and lipid secretion 6.7 times, whereas chloramphenicol inhibited their release. The formation of these vesicles from peripheral and septal wall regions did not appear to be the result of bacteriolysis. The vesicle composition of LTA and lipid was similar to that of the cytoplasmic membrane whereas the protein composition was dissimilar. The size of these vesicles ranged from 20 to 40 nm and the length of LTA ranged from 5 to 50 glycerol phosphate residues. The isolation of these vesicles provides a potential in vitro acceptor system for studying the D-alanylation of lipoteichoic acid.


Sleep bruxism: Current knowledge and contemporary management.
Bruxism is defined as the repetitive jaw muscle activity characterized by the clenching or grinding of teeth. It can be categorized into awake and sleep bruxism (SB). Frequent SB occurs in about 13% of adults. The exact etiology of SB is still unknown and probably multifactorial in nature. Current literature suggests that SB is regulated centrally (pathophysiological and psychosocial factors) and not peripherally (morphological factors). Cited consequences of SB include temporomandibular disorders, headaches, tooth wear/fracture, implant, and other restoration failure. Chairside recognition of SB involves the use of subjective reports, clinical examinations, and trial oral splints. Definitive diagnosis of SB can only be achieved using electrophysiological tools. Pharmacological, psychological, and dental strategies had been employed to manage SB. There is at present, no effective treatment that "cures" or "stops" SB permanently. Management is usually directed toward tooth/restoration protection, reduction of bruxism activity, and pain relief.


Effect of preoperative hyperinflation on static lung volumes after lung transplantation.
It is still not known whether persistent increases in functional residual capacity (FRC) and residual volume (RV) after lung transplantation are due to preexisting hyperinflation. Therefore, the aim of this study was to determine the effects of chronic lung hyperinflation on static lung volumes after heart/lung (HLT) and bilateral lung transplantation (BLT). Static lung volumes were measured in 33 patients before and at 6 month intervals for up to 3 yrs after HLT (n=25) or BLT (n=8). The preoperative diagnosis was cystic fibrosis in 25 patients and other chronic hyperinflated lung diseases in eight patients. After surgery, total lung capacity returned to predicted normal values but FRC and RV remained greater than expected for either the recipient or the donor. At 1 yr after surgery, mean+/-SD FRC and RV were 130+/-18% and 151+/-34% of the predicted values for the recipient (p<0.001), and these figures did not change significantly over time. Similar abnormalities were found in patients with and without cystic fibrosis. After transplantation for lung diseases producing chronic hyperinflation, there is a persistent increase in functional residual capacity and residual volume. This alteration is present in patients operated on for diseases developed in both childhood and adulthood and is not recovered over time. It may be due to irreversible changes in the structure of the ribcage.


Distribution of repetitive DNA sequences in eubacteria and application to fingerprinting of bacterial genomes.
Dispersed repetitive DNA sequences have been described recently in eubacteria. To assess the distribution and evolutionary conservation of two distinct prokaryotic repetitive elements, consensus oligonucleotides were used in polymerase chain reaction [PCR] amplification and slot blot hybridization experiments with genomic DNA from diverse eubacterial species. Oligonucleotides matching Repetitive Extragenic Palindromic [REP] elements and Enterobacterial Repetitive Intergenic Consensus [ERIC] sequences were synthesized and tested as opposing PCR primers in the amplification of eubacterial genomic DNA. REP and ERIC consensus oligonucleotides produced clearly resolvable bands by agarose gel electrophoresis following PCR amplification. These band patterns provided unambiguous DNA fingerprints of different eubacterial species and strains. Both REP and ERIC probes hybridized preferentially to genomic DNA from Gram-negative enteric bacteria and related species. Widespread distribution of these repetitive DNA elements in the genomes of various microorganisms should enable rapid identification of bacterial species and strains, and be useful for the analysis of prokaryotic genomes.


Kaposi's sarcoma-associated herpesvirus induction of chromosome instability in primary human endothelial cells.
Chromosome instability contributes to the multistep oncogenesis of cancer cells. Kaposi's sarcoma (KS), an angiogenic vascular spindle cancer of endothelial cells, displays stage advancement with lesions at early stage being hyperproliferative, whereas lesions at late stage are clonal or multiclonal and can exhibit a neoplastic nature and chromosome instability. Although infection with KS-associated herpesvirus (KSHV) has been associated with the initiation and promotion of KS, the mechanism of KS neoplastic transformation remains unclear. We show that KSHV infection of primary human umbilical vein endothelial cells induces abnormal mitotic spindles and centrosome duplication. As a result, KSHV-infected cells manifest chromosome instability, including chromosomal misalignments and laggings, mitotic bridges, and formation of micronuclei and multinucleation. Our results indicate that KSHV infection could predispose cells to malignant transformation through induction of genomic instability and contributes to the development of KS.


Friction and universal contact area law for randomly rough viscoelastic contacts.
We present accurate numerical results for the friction force and the contact area for a viscoelastic solid (rubber) in sliding contact with hard, randomly rough substrates. The rough surfaces are self-affine fractal with roughness over several decades in length scales. We calculate the contribution to the friction from the pulsating deformations induced by the substrate asperities. We also calculate how the area of real contact, A(v, p), depends on the sliding speed v and on the nominal contact pressure p, and we show how the contact area for any sliding speed can be obtained from a universal master curve A(p). The numerical results are found to be in good agreement with the predictions of an analytical contact mechanics theory.


Transcriptional differences between vesicular hand eczema and atopic dermatitis.
Transcriptome analyses of vesicular hand eczema (VHE) indicated a large overlap with atopic dermatitis (AD). However, differentially expressed genes (DEGs) that differentiate VHE from AD are unknown. To identify distinctive transcriptional features of VHE in comparison to AD. We re-analysed RNA sequencing data of 10 lesional palmar VHE epidermal biopsies and performed DEG analyses. We adjusted the obtained DEG results of 57 lesional whole AD skin biopsies of the upper extremities or trunk to our criteria. Up- and down-regulated DEGs in both skin diseases, VHE-only, AD-only, and opposite regulated DEGs were identified. Enrichment analyses and Chi-squared tests were conducted to test for differences in gene set enrichment between both skin diseases. Comparing 3028 DEGs in VHE (1645 up; 1383 down) with 5391 DEGs in AD (3842 up; 1549 down), revealed 1516 shared DEGs (1179 up; 337 down) and 1512 DEGs unique to VHE (466 up, 1046 down). Interferon signalling and necroptosis were significantly more prominent in VHE compared to AD. Downregulated genes identified only in VHE (like DNASE1L2, KRT2, KRT9 and KRT25) indicate an aberrant epidermal differentiation. Our study indicates a common pathophysiology between VHE and AD, but also reveals transcriptional differences between VHE and AD.


Tissue Engineered Esophageal Patch by Mesenchymal Stromal Cells: Optimization of Electrospun Patch Engineering.
Aim of work was to locate a simple, reproducible protocol for uniform seeding and optimal cellularization of biodegradable patch minimizing the risk of structural damages of patch and its contamination in long-term culture. Two seeding procedures are exploited, namely static seeding procedures on biodegradable and biocompatible patches incubated as free floating (floating conditions) or supported by CellCrownTM insert (fixed conditions) and engineered by porcine bone marrow MSCs (p-MSCs). Scaffold prototypes having specific structural features with regard to pore size, pore orientation, porosity, and pore distribution were produced using two different techniques, such as temperature-induced precipitation method and electrospinning technology. The investigation on different prototypes allowed achieving several implementations in terms of cell distribution uniformity, seeding efficiency, and cellularization timing. The cell seeding protocol in stating conditions demonstrated to be the most suitable method, as these conditions successfully improved the cellularization of polymeric patches. Furthermore, the investigation provided interesting information on patches' stability in physiological simulating experimental conditions. Considering the in vitro results, it can be stated that the in vitro protocol proposed for patches cellularization is suitable to achieve homogeneous and complete cellularizations of patch. Moreover, the protocol turned out to be simple, repeatable, and reproducible.


Adenosine 5'-triphosphate-dependent vitamin D sterol binding to heat shock protein-70 chaperones.
Chaperone proteins in the heat shock protein-70 family possess endogenous ATP binding and ATPase activity and interact with intracellular protein substrates in an ATP-dependent manner; the hydrolysis of ATP to ADP results in an increase in the affinity of the chaperone for protein substrates. Heat shock protein-70s can also specifically interact with 25-hydroxylated vitamin D metabolites. Using constitutively expressed heat shock protein-70 (hsc70) as chaperone, here we demonstrate that vitamin D metabolite binding to hsc70 is also ATP dependent. Transient overexpression of an hsc70-green fluorescent protein chimeric construct in primate kidney cells resulted in a 6-fold increase in specific, extractable 25-hydroxyvitamin D(3) binding. When ATPase capability of hsc70 was disabled, this increase was completely blocked. In solution, the binding of 25-hydroxylated vitamin D metabolites to hsc70 was significantly increased (P < 0.01) in the presence of ATP and a nonmetabolizable ATP analog. The ATP-directed increase in specific binding resulted from an increase in the abundance of relatively high-affinity hormone-binding sites (K(d), approximately 0.24 nM). These results suggest that ATP hydrolysis to ADP would favor the release of vitamin D from a donor hsc70 molecule at a time when an hsc70-bound acceptor protein substrate is anchored to the chaperone with relative avidity. We theorize that the endogenous ATPase activity of hsc70 promotes the transfer of vitamin D sterols to other intracellular vitamin D binding proteins, such as the vitamin D receptor and vitamin D hydroxylases, to which hsc70 is known to bind.


Normal range of hepatic fat fraction on dual- and triple-echo fat quantification MR in children.
To evaluate hepatic fat fraction on dual- and triple-echo gradient-recalled echo MRI sequences in healthy children. We retrospectively reviewed the records of children in a medical check-up clinic from May 2012 to November 2013. We excluded children with abnormal laboratory findings or those who were overweight. Hepatic fat fraction was measured on dual- and triple-echo sequences using 3T MRI. We compared fat fractions using the Wilcoxon signed rank test and the Bland-Altman 95% limits of agreement. The correlation between fat fractions and clinical and laboratory findings was evaluated using Spearman's correlation test, and the cut-off values of fat fractions for diagnosing fatty liver were obtained from reference intervals. In 54 children (M:F = 26:28; 5-15 years; mean 9 years), the dual fat fraction (0.1-8.0%; median 1.6%) was not different from the triple fat fraction (0.4-6.5%; median 2.7%) (p = 0.010). The dual- and triple-echo fat fractions showed good agreement using a Bland-Altman plot (-0.6 ± 2.8%). Eight children (14.8%) on dual-echo sequences and six (11.1%) on triple-echo sequences had greater than 5% fat fraction. From these children, six out of eight children on dual-echo sequences and four out of six children on triple-echo sequences had a 5-6% hepatic fat fraction. When using a cut-off value of a 6% fat fraction derived from a reference interval, only 3.7% of children were diagnosed with fatty liver. There was no significant correlation between clinical and laboratory findings with dual and triple-echo fat fractions. Dual fat fraction was not different from triple fat fraction. We suggest a cut-off value of a 6% fat fraction is more appropriate for diagnosing fatty liver on both dual- and triple-echo sequences in children.


Genome-wide association studies--data generation, storage, interpretation, and bioinformatics.
Genome-wide association studies (GWAS) have had great success in identifying common genetic determinants of disease. One of the challenges posed by GWAS is the analysis of the large amount of data generated. This review aims to provide the non-geneticists with an overview of the different steps entailed in analysis of GWAS data, with an emphasis on popular bioinformatics tools available. GWAS data generation, analysis, and interpretation will be covered.


Accessing the neural drive to muscle and translation to neurorehabilitation technologies.
This review describes methods for interfacing motor neurons from muscle recordings and their applications in studies on the neural control of movement and in the design of technologies for neurorehabilitation. After describing methods for accessing the neural drive to muscles in vivo in humans, we discuss the mechanisms of transmission of synaptic input into motor neuron output and of force generation. The synaptic input received by a motor neuron population is largely common among motor neurons. This allows linear transmission of the input and a reduced dimensionality of control by the central nervous system. Force is generated by low-pass filtering the neural signal sent to the muscle. These concepts on neural control of movement are used for the development of neurorehabilitation technologies, which are discussed with representative examples on movement replacement, restoration, and neuromodulation. It is concluded that the analysis of the output of spinal motor neurons from muscle signals provides a unique means for understanding the neural coding of movement in vivo in humans and thus for reproducing this code artificially with the aim of restoring lost or impaired motor functions.


Quaternary structure of LOV-domain containing polypeptide of Arabidopsis FKF1 protein.
Flavin-binding, Kelch repeat, F-box (FKF1) protein is a photoreceptor to regulate flowering of Arabidopsis. The protein has a light, oxygen and voltage (LOV)-sensing domain binding a flavin mononucleotide. The photo-activation of the domain is an indispensable step to initiate the cellular signaling for flowering. In the present study, a LOV-containing polypeptide of FKF1 was prepared by an overexpression system, and the quaternary structure of it was studied by size exclusion chromatography and small-angle X-ray scattering. The apparent molecular weight from chromatography suggested a globular trimeric or an anisotropic-shaped dimeric association of the polypeptide in solution. The scattering experiment demonstrated a dimeric association of the polypeptides with an elongated molecular shape displaying the radius of gyration of 27 A and the maximum dimension of 94 A. The molecular shape simulated from scattering profiles suggests an antiparallel association of the LOV domains in the dimer. Though the absorption spectrum of blue-light irradiated polypeptide was stable in the photoactivated state for a long period, the scattering profiles showed very small changes between the dark and light conditions. Based on the homologies in the amino-acid sequences and the scattering profiles, these results are discussed in connection with the structures and function of LOV domains of phototropin.


Hrs promotes ubiquitination and mediates endosomal trafficking of smoothened in Drosophila hedgehog signaling.
In Hedgehog (Hh) signaling, the seven-transmembrane protein Smoothened (Smo) acts as a signal transducer that is regulated by phosphorylation, ubiquitination, and cell surface accumulation. However, it is not clear how Smo cell surface accumulation and intracellular trafficking are regulated. Here, we demonstrate that inactivation of Hrs by deletion or RNAi accumulates Smo in the late endosome that is marked by late endosome markers. Inactivation of Hrs enhances the wing defects caused by dominant-negative Smo. We show that Hrs promotes Smo ubiquitination, deleting the ubiquitin-interacting-motif (UIM) in Hrs abolishes the ability of Hrs to regulate Smo ubiquitination. However, the UIM domain neither recognizes the ubiquitinated Smo nor directly interacts with Smo. Hrs lacking UIM domain still downregulates Smo activity even though to a less extent. We have characterized that the N-terminus of Hrs directly interacts with the PKA/CK1 phosphorylation clusters to prevent Smo phosphorylation and activation, indicating an ubiquitin-independent regulation of Smo by Hrs. Finally, we found that knockdown of Tsg101 accumulates Smo that is co-localized with Hrs and other late endosome markers. Taken together, our data indicate that Hrs mediates Smo trafficking in the late endosome by not only promoting Smo ubiquitination but also blocking Smo phosphorylation.


Forearm Pronation Osteotomy for Supination Contracture Secondary to Obstetrical Brachial Plexus Palsy: A Retrospective Cohort Study.
Obstetrical brachial plexus palsy can lead to fixed forearm supination contracture. Fixed supination may lead to functional deficits as the affected hand cannot be positioned optimally for activities on a desk such as writing and typing, or for using tools including utensils, which require a neutral or pronated forearm. Forearm pronation osteotomy has been used to address this problem, although the functional benefit over nonoperative management has not been clearly defined. Potentially deleterious consequences on hand function that requires supination or fine motor skills are also uncertain. Patients with fixed forearm supination contracture were selected from our institutional brachial plexus database. Those who underwent both bone forearm rotational osteotomy were analyzed for age at time of surgery, preoperative forearm resting position, active and passive supination and pronation, and preoperative function assessed by the brachial plexus outcome measure (BPOM) and active movement scale (AMS). Preoperative results were compared with values obtained at follow-up at least 12 months postoperatively. A matched cohort of children with fixed forearm supination contracture that were treated nonoperatively and followed for at least 12 months, was also selected. For this group, forearm resting position, movement, AMS, and BPOM scores were analyzed at a baseline clinic visit and the most recent follow-up. Changes in forearm resting position, AMS, and BPOM activity scale scores were then compared between groups. Records were obtained for 14 cases and 10 controls. Study groups were similar with respect to resting forearm position, hand function, and time from initial to final evaluation. Groups differed with respect to age and active supination. We observed a statistically significant change in resting position among operative patients compared with their preoperative status and compared with controls. Hand-specific AMS score did not change significantly in the operative group as compared with controls. The BPOM score for drums, reflective of function in neutral rotation to mild pronation, improved in the operated patients as compared with controls. There was no loss of plate holding ability (reflective of supination function, putty (grasp), or bead placement (fine motor) among the operated patients as compared with controls. By pronating resting forearm position by about 90 degrees to near neutral, osteotomy resulted in improved neutral to mild pronation-dependent function without loss of supination-dependent function or hand motor skills. Level III-retrospective cohort study.


Long- but not short-term adult-onset, isolated GH deficiency in male mice leads to deterioration of β-cell function, which cannot be accounted for by changes in β-cell mass.
Developmental models of GH deficiency (GHD) and excess indicate that GH is positively associated with β-cell mass. Therefore, the reduction in GH levels observed with age and weight gain may contribute to the age-related decline in β-cell function. To test this hypothesis, β-cell mass and function were assessed in a mouse model of adult-onset, isolated GHD (AOiGHD). β-Cell mass did not differ between low-fat (LF)-fed AOiGHD and controls. However, high fat-fed AOiGHD mice displayed impaired expansion of β-cell mass and a reduction of bromodeoxyuridine-labeled islet cells, whereas in vitro β-cell function (basal and glucose-stimulated insulin secretion [GSIS]) did not differ from controls. In contrast, duration of AOiGHD differentially altered in vitro β-cell function in LF-fed mice. Specifically, islets from young LF-fed AOiGHD mice showed significant reductions in insulin content and basal insulin secretion, but GSIS was similar to that of controls. A similar islet phenotype was observed in a developmental model of isolated GHD (GH-releasing hormone knockout). Given that LF- and high fat-fed AOiGHD mice, as well as GH-releasing hormone knockout mice, display improved insulin sensitivity, islet changes may be due to reduced insulin demand, rather than primary β-cell dysfunction. However, islets from older LF-fed AOiGHD mice exhibited impaired GSIS, associated with reduced expression of genes important to maintain glucose sensing, suggesting that factors secondary to AOiGHD can alter β-cell function with age. AOiGHD mice exhibited postprandial hypertriglyceridemia and increased pancreatic expression of lipid/inflammatory stress response genes (activating transcription factor 3 and peroxisome proliferator activator receptor β/δ). Therefore, we speculate that these changes may initially protect the AOiGHD β-cell, but with age, lipotoxicity may impair β-cell function.


Impact of a critical pathway on inpatient management of diabetic ketoacidosis.
To assess the management of diabetic ketoacidosis (DKA) and evaluate if introduction of a critical pathway improves management, we studied adults admitted with DKA to the Medicine and Critical Care Services in a US teaching hospital. Patients admitted with DKA in 1997 before implementation of the critical pathway were the control group (n=72). In 1998, housestaff and nurses in the emergency department (ED) and on the General Medicine and Critical Care Services were instructed in the use of the critical pathway. Patients admitted with DKA during 1998 (n=77) were the intervention group. Length of stay (LOS), hospital cost, adherence to guidelines, and medical outcomes to be avoided were compared, and regression analyses were performed to correlate processes and outcomes of care. Mean LOS and variability in LOS decreased during the intervention period, especially in patients treated without endocrinology consultation (EC) (5.2 +/- 10.6 vs. 2.4 +/- 2.1 days, P=0.01), and hospital cost and variability in cost tended to decrease ($6441 +/- 15,204 vs. $3625 +/- 3478, P=0.24). More intervention subjects received the recommended intravenous fluid volume (88 vs. 71%, P=0.013), education in sick-day management (77 vs. 54%, P=0.006), and EC (38 vs. 21%, P=0.03). Insulin management was not changed. We conclude that implementation of a DKA critical pathway reduced practice variation and was associated with shorter LOS and a trend toward decreased cost. Some processes of care were improved but many require additional interventions.


Mechanism of regulation of bcl-2 mRNA by nucleolin and A+U-rich element-binding factor 1 (AUF1).
The antiapoptotic Bcl-2 protein is overexpressed in a variety of cancers, particularly leukemias. In some cell types this is the result of enhanced stability of bcl-2 mRNA, which is controlled by elements in its 3'-untranslated region. Nucleolin is one of the proteins that binds to bcl-2 mRNA, thereby increasing its half-life. Here, we examined the site on the bcl-2 3'-untranslated region that is bound by nucleolin as well as the protein binding domains important for bcl-2 mRNA recognition. RNase footprinting and RNA fragment binding assays demonstrated that nucleolin binds to a 40-nucleotide region at the 5' end of the 136-nucleotide bcl-2 AU-rich element (ARE(bcl-2)). The first two RNA binding domains of nucleolin were sufficient for high affinity binding to ARE(bcl-2). In RNA decay assays, ARE(bcl-2) transcripts were protected from exosomal decay by the addition of nucleolin. AUF1 has been shown to recruit the exosome to mRNAs. When MV-4-11 cell extracts were immunodepleted of AUF1, the rate of decay of ARE(bcl-2) transcripts was reduced, indicating that nucleolin and AUF1 have opposing roles in bcl-2 mRNA turnover. When the function of nucleolin in MV-4-11 cells was impaired by treatment with the nucleolin-targeting aptamer AS1411, association of AUF1 with bcl-2 mRNA was increased. This suggests that the degradation of bcl-2 mRNA induced by AS1411 results from both interference with nucleolin protection of bcl-2 mRNA and recruitment of the exosome by AUF1. Based on our findings, we propose a model that illustrates the opposing roles of nucleolin and AUF1 in regulating bcl-2 mRNA stability.


Smartphone mobile application delivering personalized, real-time sun protection advice: a randomized clinical trial.
Mobile smartphones are rapidly emerging as an effective means of communicating with many Americans. Using mobile applications (apps), they can access remote databases, track time and location, and integrate user input to provide tailored health information. A smartphone mobile app providing personalized, real-time sun protection advice was evaluated in a randomized clinical trial. The trial was conducted in 2012 and had a randomized pretest-posttest controlled design with a 10-week follow-up. Data were collected from a nationwide population-based survey panel. A sample of 604 non-Hispanic and Hispanic adults from the Knowledge Panel 18 years or older who owned an Android smartphone were enrolled. The mobile app provided advice on sun protection (ie, protection practices and risk of sunburn) and alerts (to apply or reapply sunscreen and get out of the sun), hourly UV Index, and vitamin D production based on the forecast UV Index, the phone's time and location, and user input. Percentage of days using sun protection and time spent outdoors (days and minutes) in the midday sun and number of sunburns in the past 3 months were collected. Individuals in the treatment group reported more shade use (mean days staying in the shade, 41.0% vs 33.7%; P = .03) but less sunscreen use (mean days, 28.6% vs 34.5%; P = .048) than controls. There was no significant difference in number of sunburns in the past 3 months (mean, 0.60 in the treatment group vs 0.62 for controls; P = .87). Those who used the mobile app reported spending less time in the sun (mean days keeping time in the sun to a minimum, 60.4% for app users vs 49.3% for nonusers; P = .04) and using all protection behaviors combined more (mean days, 39.4% vs 33.8%; P = .04). The mobile app improved some sun protection. Use of the mobile app was lower than expected but associated with increased sun protection. Providing personalized advice when and where people are in the sun may help reduce sun exposure.


Comparison of the informative value of electrocardiographic criteria of myocardial infarction with the use of different lead systems.
The authors compared the informative value of 39 criteria of myocardial infarction with ECG registration using the standard 12-lead system (ECG-12), 24 criteria according to Frank's orthogonal system (OECG-F), and 32 criteria according to the new 4-lead system (ECG-4). 127 patients with myocardial infarction (MI) were examined. The sensitivity of the most informative ECG-4 criteria of MI was higher (44.4%) than OECG-F (39.4%) and ECG-12 (38.6%). With the application of quantitative ECG-4 criteria, signs of MI were detected in all cases diagnosed by experts using the ECG-12 system. In view of the considerable simplicity of its registration technique, the ECG-4 system can be recommended for the mass screening ECG examinations aimed especially at detection of MI.


Smooth side-match classified vector quantizer with variable block size.
Although the side-match vector quantizer (SMVQ) reduces the bit rate, the image coding quality by SMVQ generally degenerates as the gray level transition across the boundaries of the neighboring blocks is increasing or decreasing. This study presents a smooth side-match method to select a state codebook according to the smoothness of the gray levels between neighboring blocks. This method achieves a higher PSNR and better visual perception than SMVQ does for the same bit rate. Moreover, to design codebooks, a genetic clustering algorithm that automatically finds the appropriate number of clusters is proposed. The proposed smooth side-match classified vector quantizer (SSM-CVQ) is thus a combination of three techniques: the classified vector quantization, the variable block size segmentation and the smooth side-match method. Experimental results indicate that SSM-CVQ has a higher PSNR and a lower bit rate than other methods. Furthermore, the Lena image can be coded by SSM-CVQ with 0.172 bpp and 32.49 dB in PSNR.


Aquaphotomics Research of Cold Stress in Soybean Cultivars with Different Stress Tolerance Ability: Early Detection of Cold Stress Response.
The development of non-destructive methods for early detection of cold stress of plants and the identification of cold-tolerant cultivars is highly needed in crop breeding programs. Current methods are either destructive, time-consuming or imprecise. In this study, soybean leaves' spectra were acquired in the near infrared (NIR) range (588-1025 nm) from five cultivars genetically engineered to have different levels of cold stress tolerance. The spectra were acquired at the optimal growing temperature 27 °C and when the temperature was decreased to 22 °C. In this paper, we report the results of the aquaphotomics analysis performed with the objective of understanding the role of the water molecular system in the early cold stress response of all cultivars. The raw spectra and the results of Principal Component Analysis, Soft Independent Modeling of Class Analogies and aquagrams showed consistent evidence of huge differences in the NIR spectral profiles of all cultivars under normal and mild cold stress conditions. The SIMCA discrimination between the plants before and after stress was achieved with 100% accuracy. The interpretation of spectral patterns before and after cold stress revealed major changes in the water molecular structure of the soybean leaves, altered carbohydrate and oxidative metabolism. Specific water molecular structures in the leaves of soybean cultivars were found to be highly sensitive to the temperature, showing their crucial role in the cold stress response. The results also indicated the existence of differences in the cold stress response of different cultivars, which will be a topic of further research.


[Reaction of endonuclease EcoRI with DNA of the plasmid ColEI].
Interaction of restrictase EcoRI with supercoiled and open forms of ColEI DNA is studied in the range 0-55 degrees. At temperatures 25-37 degrees superhelical ColEI DNA is converted to linear form, the most complete cleavage of supercoiled molecules were observed at 25 degrees. This reaction is not followed by accumulation of intermediate open circular DNA forms. At 45-55 degrees open circular DNA forms with nicks at the restriction site are formed along with linear molecules. Similar effects were observed at 10 degrees, whereas at 0 degree only open circular DNA molecules are formed.


Water hydrogen-bonding structure and dynamics near lipid multibilayer surface: Molecular dynamics simulation study with direct experimental comparison.
Lipid multibilayers are excellent model systems for studying water structures and dynamics near biological membrane surfaces. In particular, the orientational distribution and rotational dynamics of water molecules near hydrophilic lipid groups are found to be sensitive to the chemical nature and charge distributions of the amphiphilic lipids. To elucidate how different parts of these lipids affect the water hydrogen-bonding structure and dynamics and to directly compare with recent experimental results, we carried out molecular dynamics (MD) simulations of lipid multibilayer systems. We found that the water molecules close to positively charged choline groups have a broad distribution of orientations due to the clathratelike shell formation around the choline groups but that those associated with phosphate groups, even in the second hydration shell, are orientationally restricted due to their strong hydrogen bonding with the phosphate group. These MD simulation results are in excellent agreement with our time-resolved infrared pump-probe anisotropy measurements, and we believe that they provide valuable insights into the role of water molecules in maintaining lipid bilayer integrity.


Antibodies for treating and preventing disease: the potential role of polymeric controlled release.
Following the identification of antibodies (Abs) as agents of immunity, it was hypothesized that individuals could be both (1) protected against disease by the transfer of unmodified Ab (passive immunization), and (2) cured of established disease by Ab armed with cytotoxic agents (immunotherapy). The development of monoclonal antibody (mAb) technology in 1975 reinvigorated these ideas. Although passive immunization has been practiced with great success for many years, successful tissue targeting by systemically delivered immunotoxins in humans has been documented in only a few cases. New modes of drug delivery, engineered for mAb-based products, may enable new applications of passive immunization and may provide improved tissue targeting for immunotherapy. By allowing sustained and tissue-localized delivery of mAb-conjugates, controlled-release polymers may play an important role in this effort. This article reviews the use of mAb in treating and preventing human disease, as well as the pharmacokinetics of Ab delivery. Two areas where controlled Ab release may yield new therapies are highlighted: sustained passive immunization of the mucus secretions and immunotherapy of brain tumors.


Pathogenicity of avian malaria in experimentally-infected Hawaii Amakihi.
The introduction of avian malaria (Plasmodium relictum) and mosquitoes (Culex quinquefasciatus) to the Hawaiian Islands (USA) is believed to have played a major role in the decline and extinction of native Hawaiian honeycreepers (Drepanidinae). This introduced disease is thought to be one of the primary factors limiting recovery of honeycreepers at elevations below 1,200 m where native forest habitats are still relatively intact. One of the few remaining species of honeycreepers with a wide elevational distribution is the Hawaii Amakihi (Hernignathus virens). We measured morbidity and mortality in experimentally-infected Hawaii Amakihi that were captured in a high elevation, xeric habitat that is above the current range of the mosquito vector. Mortality among amakihi exposed to a single infective mosquito bite was 65% (13/20). All infected birds had significant declines in food consumption and a corresponding loss in body weight over the 60 day course of the experiment. Gross and microscopic lesions in birds that succumbed to malaria included enlargement and discoloration of the spleen and liver and parasitemias as high as 50% of circulating erythrocytes. Mortality in experimentally-infected amakihi was similar to that observed in Apapane (Himnatione sanguinea) and lower than that observed in Iiwi (Vestiaria coccinea) infected under similar conditions with the same parasite isolate. We conclude that the current elevational and geographic distribution of Hawaiian honeycreepers is determined by relative susceptibility to avian malaria.


Targeting the Aryl Hydrocarbon Receptor with Microbial Metabolite Mimics Alleviates Experimental Colitis in Mice.
Targeting the aryl hydrocarbon receptor (AhR) is an emerging therapeutic strategy for multiple diseases (e.g., inflammatory bowel disease). Thermosporothrix hazakensis microbial metabolite 2-(1'H-indole-3'-carbonyl)-thiazole-4-carboxylic acid methyl ester (ITE) is a putative AhR endogenous ligand. To improve the chemical stability, we synthesized a series of ITE chemical mimics. Using a series of in vitro assays, we identified 2-(1H-indole-3-carbonyl)-N-methyl thiazole-4-carboxamide (ITE-CONHCH3) as a highly potent (EC50 = 1.6 nM) AhR agonist with high affinity (Ki = 88 nM). ITE-CONHCH3 triggered AhR nuclear translocation and dimerization of AhR-ARNT, enhanced AhR binding in the CYP1A1 promoter, and induced AhR-regulated genes in an AhR-dependent manner. The metabolic stability of ITE-CONHCH3 in a cell culture was 10 times higher than that of ITE. Finally, we observed protective effects of ITE-CONHCH3 in mice with DSS-induced colitis. Overall, we demonstrate and validate a concept of microbial metabolite mimicry in the therapeutic targeting of AhR.


Efficient differentiation of vascular smooth muscle cells from Wharton's Jelly mesenchymal stromal cells using human platelet lysate: A potential cell source for small blood vessel engineering.
The development of fully functional small diameter vascular grafts requires both a properly defined vessel conduit and tissue-specific cellular populations. Mesenchymal stromal cells (MSCs) derived from the Wharton's Jelly (WJ) tissue can be used as a source for obtaining vascular smooth muscle cells (VSMCs), while the human umbilical arteries (hUAs) can serve as a scaffold for blood vessel engineering. To develop VSMCs from WJ-MSCs utilizing umbilical cord blood platelet lysate. WJ-MSCs were isolated and expanded until passage (P) 4. WJ-MSCs were properly defined according to the criteria of the International Society for Cell and Gene Therapy. Then, these cells were differentiated into VSMCs with the use of platelet lysate from umbilical cord blood in combination with ascorbic acid, followed by evaluation at the gene and protein levels. Specifically, gene expression profile analysis of VSMCs for ACTA2, MYH11, TGLN, MYOCD, SOX9, NANOG homeobox, OCT4 and GAPDH, was performed. In addition, immunofluorescence against ACTA2 and MYH11 in combination with DAPI staining was also performed in VSMCs. HUAs were decellularized and served as scaffolds for possible repopulation by VSMCs. Histological and biochemical analyses were performed in repopulated hUAs. WJ-MSCs exhibited fibroblastic morphology, successfully differentiating into "osteocytes", "adipocytes" and "chondrocytes", and were characterized by positive expression (> 90%) of CD90, CD73 and CD105. In addition, WJ-MSCs were successfully differentiated into VSMCs with the proposed differentiation protocol. VSMCs successfully expressed ACTA2, MYH11, MYOCD, TGLN and SOX9. Immunofluorescence results indicated the expression of ACTA2 and MYH11 in VSMCs. In order to determine the functionality of VSMCs, hUAs were isolated and decellularized. Based on histological analysis, decellularized hUAs were free of any cellular or nuclear materials, while their extracellular matrix retained intact. Then, repopulation of decellularized hUAs with VSMCs was performed for 3 wk. Decellularized hUAs were repopulated efficiently by the VSMCs. Biochemical analysis revealed the increase of total hydroyproline and sGAG contents in repopulated hUAs with VSMCs. Specifically, total hydroxyproline and sGAG content after the 1st, 2nd and 3rd wk was 71 ± 10, 74 ± 9 and 86 ± 8 μg hydroxyproline/mg of dry tissue weight and 2 ± 1, 3 ± 1 and 3 ± 1 μg sGAG/mg of dry tissue weight, respectively. Statistically significant differences were observed between all study groups (P < 0.05). VSMCs were successfully obtained from WJ-MSCs with the proposed differentiation protocol. Furthermore, hUAs were efficiently repopulated by VSMCs. Differentiated VSMCs from WJ-MSCs could provide an alternative source of cells for vascular tissue engineering.


Socio-demographic parameters in screening for breast cancer: Lessons from a population-'based women's Health Project held in a province in Turkey.
to inform healthy women about breast cancer and screen them, as well as to look for any relationship between demographic and clinical findings and breast cancer. thirty-five health teams were created prior to the study. The teams were primarily trained for breast examination and for screening methods to detect breast cancer. a total of 77,934 subjects were evaluated. Clinical breast examination (CBE) was performed in 66% (n=51,706) of the participants. The characteristics of the subjects in the examined group were similar to those in the group refusing examination. The percentage of the subjects who declined examination was 2-fold higher in the ≥ 60 year age group compared to younger women. A breast mass was detected in 2,838 (6%) subjects who had undergone breast examination. Lower educational level and urban dwellers showed higher incidence of suspicious mass in CBE. Fifty-eight women were diagnosed with breast cancer. The cumulative incidence of breast cancer was 7.5/10.000 for all of the study population and 10.1/10.000 for women with CBE. elderly subjects, those living in rural areas and women with low educational and lower socio-economic levels should be convinced to undergo screening for breast cancer.


Tunable sieving of ions using graphene oxide membranes.
Graphene oxide membranes show exceptional molecular permeation properties, with promise for many applications. However, their use in ion sieving and desalination technologies is limited by a permeation cutoff of ∼9 Å (ref. 4), which is larger than the diameters of hydrated ions of common salts. The cutoff is determined by the interlayer spacing (d) of ∼13.5 Å, typical for graphene oxide laminates that swell in water. Achieving smaller d for the laminates immersed in water has proved to be a challenge. Here, we describe how to control d by physical confinement and achieve accurate and tunable ion sieving. Membranes with d from ∼9.8 Å to 6.4 Å are demonstrated, providing a sieve size smaller than the diameters of hydrated ions. In this regime, ion permeation is found to be thermally activated with energy barriers of ∼10-100 kJ mol-1 depending on d. Importantly, permeation rates decrease exponentially with decreasing sieve size but water transport is weakly affected (by a factor of <2). The latter is attributed to a low barrier for the entry of water molecules and large slip lengths inside graphene capillaries. Building on these findings, we demonstrate a simple scalable method to obtain graphene-based membranes with limited swelling, which exhibit 97% rejection for NaCl.


An experimental study on arsoles: structural variation, optical and electronic properties, and emission behavior.
We experimentally demonstrated the intrinsic nature of arsoles as promising functional heteroles. A series of 2,5-diarylarsoles are easily and safely prepared through the procedure in which non-volatile arsenic intermediates are employed to overcome the synthetic barrier due to the concern of volatility of the arsenic precursors used in conventional methods. A Pd-catalyzed Suzuki-Miyaura coupling reaction can be applied to the obtained arsoles for fine molecular design, unlikely to phospholes. It was demonstrated that the optical and electronic properties, i.e. emission colors, quantum yields, and energy levels of the frontier orbitals, are similar to those of phospholes, as conventional theoretical studies have predicted. Furthermore, it was found that arsoles showed mechanochromic properties.


Mental models: a basic concept for human factors design in infection prevention.
Much of the effort devoted to promoting better hand hygiene is based on the belief that poor hand hygiene reflects poor motivation. We argue, however, that automatic unconscious behaviour driven by 'mental models' is an important contributor to what actually happens. Mental models are concepts of reality--imaginary, often blurred, and sometimes unstable. Human beings use them to reduce mental load and free up capacity in the conscious mind to focus on deliberate activities. They are pragmatic solutions to the complexity of life. Knowledge of such mental processes helps healthcare designers and clinicians overcome barriers to behavioural change. This article reviews the concept of mental models and considers how it can be used to improve hand hygiene and patient safety.


Non-Hodgkin's lymphoma involving the vagina: a clinicopathologic analysis of 14 patients.
Non-Hodgkin's lymphomas (NHL) uncommonly involve the vagina. In this study, 14 NHL involving the vagina are reported. Eight cases were stage IE or IIE and are presumed to be primary. The mean age of these eight patients at presentation was 42 years (range, 26-66 yrs), and four of eight patients complained primarily of vaginal bleeding. Histologically, all eight neoplasms were diffuse large B-cell lymphoma (DLBCL). Clinical follow up ranged from 1.8 to 18 years. Six of eight patients were alive without evidence of disease at the last follow up (range, 2.8-21 yrs), one patient died of unrelated causes at 9 years, and one patient died from NHL at 1.8 years. In six patients vaginal involvement was part of systemic disease at diagnosis, either stage IIIE or IV. The mean patient age at the time vaginal involvement was detected was 65 years (range, 49-82 yrs). Four of six patients had vaginal bleeding. Five neoplasms were DLBCL and one tumor was B-cell small lymphocytic lymphoma/chronic lymphocytic leukemia. Clinical follow up for these patients ranged from 2 weeks to 13 years. Two patients were free of disease after treatment at 4.5 and 13 years, two patients were alive with progressive NHL, one patient died of NHL, and one patient was recently diagnosed. The authors conclude that low-stage (presumably primary) vaginal NHL are DLBCL, tend to occur in younger women, and cause vaginal bleeding. High-stage NHL involving the vagina are usually DLBCL, tend to affect older women, and are relatively more heterogeneous clinically and histologically, but also usually cause vaginal bleeding.


Dimensionality reduction in neural models: an information-theoretic generalization of spike-triggered average and covariance analysis.
We describe an information-theoretic framework for fitting neural spike responses with a Linear-Nonlinear-Poisson cascade model. This framework unifies the spike-triggered average (STA) and spike-triggered covariance (STC) approaches to neural characterization and recovers a set of linear filters that maximize mean and variance-dependent information between stimuli and spike responses. The resulting approach has several useful properties, namely, (1) it recovers a set of linear filters sorted according to their informativeness about the neural response; (2) it is both computationally efficient and robust, allowing recovery of multiple linear filters from a data set of relatively modest size; (3) it provides an explicit "default" model of the nonlinear stage mapping the filter responses to spike rate, in the form of a ratio of Gaussians; (4) it is equivalent to maximum likelihood estimation of this default model but also converges to the correct filter estimates whenever the conditions for the consistency of STA or STC analysis are met; and (5) it can be augmented with additional constraints on the filters, such as space-time separability. We demonstrate the effectiveness of the method by applying it to simulated responses of a Hodgkin-Huxley neuron and the recorded extracellular responses of macaque retinal ganglion cells and V1 cells.


The effects of methanolic extracts of ginger (Zingiber officinale) on human sperm parameters; an in vitro study.
This study was conducted on Zingiber officinale or ginger on motility, grading and morphological aspects of human sperm. Thirty human semen samples were obtained from the local hospital. The samples were swim up by Ham's F10. The samples (0.9 mL) were treated by 0.1 mL of ginger methanolic extracts (0.1, 0.2, 0.4 and 0.6% concentration). Sperm motility, grading and morphology parameters were assessed using light microscope at 0, 2, 4, 8 and 24 h after treatment. Dose and time-dependent decreases in motility accompanied by concomitant decrease in grading 3 and 4 were noticed. Morphologic profiles of the sperms were changed under different doses of ginger on the basis of time of assess. These data indicate that some exhibits a lower percentage of motility and grading when methanolic ginger is added to semen fluid. According to the results it would be concluded that ginger can induce the toxic effects on sperm parameters.


HBV Drug Resistance Substitutions Existed before the Clinical Approval of Nucleos(t)ide Analogues: A Bioinformatic Analysis by GenBank Data Mining.
Naturally occurring nucleos(t)ide analogue resistance (NUCr) substitution frequencies in the reverse transcriptase (RT) of the hepatitis B virus (HBV) were studied extensively after the clinical approval of nucleos(t)ide analogues (NUCs; year of approval 1998). We aimed to study NUCr substitutions in HBV RT sequences obtained before 1998 and better understand the evolution of RT sequences without NUC pressures. Our strategy was to retrieve HBV sequences from GenBank deposited before 1998. The initial search used the keywords "hepatitis B virus" or "HBV" and 1139 sequences were found. Data analyses included information extraction: sequence quality control and amino acid substitution analysis on 8 primary NUCr and 3 secondary substitution codons. Three hundred and ninety-four RT-containing sequences of 8 genotypes from 25 countries in 4 continents were selected. Twenty-seven (6.9%) sequences were found to harbor substitutions at NUCr-related codons. Secondary substitutions (rtL80V and rtV173G/A/L) occurred more frequently than primary NUCr substitutions (rtI169L; rtA181G; T184A/S; rtS202T/R; rtM204L and rtM250K). Typical amino acid substitutions associated with NUCr were of rtL80V, rtV173L and rtT184A/S. We confirm the presence of naturally occurring typical HBV NUCr substitutions with very low frequencies, and secondary substitutions are more likely to occur than primary NUCr substitutions without the selective pressure of NUCs.


Vasculature-on-a-Chip with a Porous Membrane Electrode for In Situ Electrochemical Detection of Nitric Oxide Released from Endothelial Cells.
Vasculature-on-a-chip is a microfluidic cell culture device used for modeling vascular functions by culturing endothelial cells. Porous membranes are widely used to create cell culture environments. However, in situ real-time measurements of cellular metabolites in microchannels are challenging. In this study, a novel microfluidic device with a porous membrane electrode was developed for the in situ monitoring of nitric oxide (NO) released by endothelial cells in real time. In this system, a porous Au membrane electrode was placed directly beneath the cells for in situ and real-time measurements of NO, a biomarker of endothelial cells. First, the device was electrochemically characterized to construct a calibration plot for NO. Next, NO released by human umbilical vein endothelial cells under l-arginine stimulation was successfully quantified. Furthermore, the changes in NO release with culture time (in days) using the same sample were successfully recorded by exploiting minimally invasive measurements. This is the first report on the combination of a microfluidic device and porous membrane electrode for the electrochemical analysis of endothelial cells. This device will contribute to the development of organ-on-a-chip technology for real-time in situ cell analyses.


Blood level of mitochondrial aspartate aminotransferase as an indicator of the extent of ischemic necrosis of the rat liver.
To assess the severity of ischemic liver injury, we examined release of mitochondrial aspartate aminotransferase (EC 126.96.36.199) and its cytoplasmic isozyme from the ischemic rat liver into the circulation. Their patterns of leakage were quite different: the level of cytoplasmic aspartate aminotransferase reached a peak soon after the circulation to the ischemic liver was restored, while that of mitochondrial aspartate aminotransferase increased slowly, reaching a maximum after more than 10 hr. On anoxic incubation of mitochondria isolated from the normal liver, oxidative phosphorylation capacity was lost within 2 hr, at which time no leakage of matrix enzymes was observed: more than 10 hr after-loss-of-oxidative phosphorylation were needed for the matrix enzymes to leak out of the mitochondrial membrane. Since the viability of cells is considered to depend on the capacity of oxidative phosphorylation, it is highly likely that the delayed appearance of mitochondrial aspartate aminotransferase in blood indicates the postmortem changes of injured cells. In fact, the cumulative activity of mitochondrial aspartate aminotransferase but not cytoplasmic aspartate aminotransferase in circulation after ischemic liver injury correlated fairly well with the decrease of total adenine nucleotides which were monitored to measure viable cells. The difference between mitochondrial aspartate aminotransferase and cytoplasmic aspartate aminotransferase as quantitative indices of hepatic necrosis may be due to the relative stability of the former and significant inactivation of the latter during hepatic ischemia. Therefore, the determination of mitochondrial aspartate aminotransferase in blood may be useful in the assessment of liver necrosis after ischemic injury.


Ambient-Air-Stable Lead-Free CsSnI3 Solar Cells with Greater than 7.5% Efficiency.
Black orthorhombic (B-γ) CsSnI3 with reduced biotoxicity and environmental impact and excellent optoelectronic properties is being considered as a promising eco-friendly candidate for high-performing perovskite solar cells (PSCs). A major challenge in a large-scale implementation of CsSnI3 PSCs includes the rapid transformation of Sn2+ to Sn4+ (within a few minutes) under an ambient-air condition. Here, we demonstrate that ambient-air stable B-γ CsSnI3 PSCs can be fabricated by incorporating N,N'-methylenebis(acrylamide) (MBAA) into the perovskite layer and by using poly(3-hexylthiophene) as the hole transporting material. The lone electron pairs of -NH and -CO units of MBAA are designed to form coordination bonding with Sn2+ in the B-γ CsSnI3, resulting in a reduced defect (Sn4+) density and better stability under multiple conditions for the perovskite light absorber. After a modification, the highest power conversion efficiency (PCE) of 7.50% is documented under an ambient-air condition for the unencapsulated CsSnI3-MBAA PSC. Furthermore, the MBAA-modified devices sustain 60.2%, 76.5%, and 58.4% of their initial PCEs after 1440 h of storage in an inert condition, after 120 h of storage in an ambient-air condition, and after 120 h of 1 Sun continuous illumination, respectively.


Politics Spread COVID: Developing a Public Health Response.
The circumstances leading to one million American deaths from COVID-19 are familiar to health educators: The pandemic was politicized from the outset; public health professionals were pushed aside and sometimes attacked; in many areas, compliance with public health recommendations was low, and vaccine uptake was much less than required to meet the threat; the public health community tied itself in knots trying to figure out how to cut through the plethora of misinformation; people in marginalized populations died in vastly disproportionate numbers in spite of years of preparation to prevent just that outcome. Cumulative mortality is equivalent to some of the "worst case" scenarios put forth by U.S. public health experts at the beginning of the pandemic even though we've worked so hard to prepare for this type of global pandemic, so what went wrong? Profound changes in American politics have led to a relationship between public health and swaths of society that is quite unlike what previously was assumed in the dominant models of public health; it was believed that public health experts would be treated as and listened to as the experts they are in the field. As the politicization of the pandemic and subsequent deaths show, these assumptions are no longer valid and we cannot assure the health of the public as we are required to do. The assumptions that we have operated under for so long in public health now must be deconstructed and revisited in order to move forward and prevent unnecessary future deaths. To do this, we must better understand the influence of American politics and we must more effectively engage in politics at all levels.


Development of a simple recovery-enrichment system for enhanced detection of heat-injured Listeria monocytogenes in pasteurized milk.
A simple anaerobic recovery-enrichment system, semisolid Penn State University (ssPSU) broth, that enhances recovery of heat-injured Listeria monocytogenes, was rapidly achieved in 10-ml screw-capped tubes by adding Bacto-agar (2.5 g/liter) and L-cysteine (0.5 g/liter) to Penn State University broth. Glucose was removed from the formulation for ssPSU broth to prevent the growth of thermoduric lactobacilli. Ferric ammonium citrate was added to ssPSU broth to detect esculin hydrolysis and to indicate the presumptive presence of L. monocytogenes. Replacement of phosphate buffer with 3-[N-morpholino]propanesulfonic acid (MOPS) buffer and addition of magnesium sulfate (15 mM) enhanced recovery and detection of L. monocytogenes heat treated at 62.8 degrees C for 20 min. D-Serine, at a concentration of 150 mM, was found to inhibit germination of Bacillus spp. spores but did not inhibit severely heat-injured L. monocytogenes. Finally, ssPSU broth was modified (to mPSU broth) to contain the following: (i) Bacto-agar, 2.5 g/liter; (ii) ferric ammonium citrate, 0.5 g/liter; (iii) MOPS buffer, pH 7.0; (iv) D-serine, 13.7 g/liter; (v) D-alanine, 11.6 g/liter; and (iv) magnesium sulfate, 1.81 g/liter. Incubation temperature significantly affected the recovery and detection of severely heat-injured L. monocytogenes. L. monocytogenes that were heat challenged in filter-sterilized whole milk at 62.8 degrees C for 20, 25, and 30 min could not be detected at incubation temperatures > or = 30 degrees C but were consistently detected after incubation at 25 degrees C for 174, 199, and 330 h, respectively. Heat-injured cells of L. monocytogenes that were added to various commercial brands of pasteurized whole milk were also detected using mPSU broth. When clostridial spores (10(4) spores per ml) were added to filter-sterilized milk containing either heat-injured or non-heat-injured L. monocytogenes, only the latter could be detected in mPSU broth. The mPSU broth system requires no purging with nitrogen gas to create anaerobic conditions and permits recovery, growth, and detection of L. monocytogenes in one vessel in the presence of thermoduric background microflora commonly found in pasteurized milk.


Clinical and experimental pharmacokinetic interaction between 6-mercaptopurine and methotrexate.
Clinical and experimental pharmacokinetic interaction between 6-mercaptopurine (6-MP) and methotrexate (MTX) was investigated in patients as well as in rats and in HL-60 human leukemic cells. Ten children affected by acute lymphoblastic leukemia (ALL) in remission received daily doses of 6-MP given at 25 mg/m2 and i.v. infusion of high-dose MTX at 2 or 5 g/m2 once every other week. When 6-MP was given alone, the mean peak plasma concentration (Cmax) and area under the curve (AUC) of 6-MP were 72.5 ng/ml and 225.3 h ng ml(-1). Concurrent treatment with MTX at 2 or 5 g/m2 resulted in a mean increase of 108% and 121% in the Cmax and of 69% and 93% in the AUC, respectively. In rats treated with an oral dose of 6-MP at 75 mg/m2, MTX given i.p. at 5 g/m2 produced mean increases of 110% and 230% in the Cmax and AUC of 6-MP, respectively. In HL-60 human leukemic cells incubated with 6-MP at 250 ng/ml, the cumulative intracellular concentration of 6-thioguanine and 6-MP nucleotides was not significantly modified by treatment with 20 micrograms/ml of MTX. The present findings indicate that high-dose MTX enhances the bioavailability of 6-MP as evidenced by the observed increases in the plasma Cmax and AUC of 6-MP in humans and animals.


Effects of temperature on the performance of finishing swine: II. Effects of a cold, diurnal temperature on average daily gain, feed intake, and feed efficiency.
Forty-eight crossbred barrows and gilts weighing 84.5 +/- .33 kg were used during two 21-d trials to investigate the effects of a cold, diurnal temperature (CD; -5.0 to 8.0 degrees C) compared with a constant, thermoneutral temperature (TN; 20 degrees C) and the effects of sex (barrows vs gilts) on performance. A second objective was to determine shrinkage as a result of a 24-h fast immediately after the 21-d study of hogs commingled vs those not commingled for both environmental treatments (CD vs TN). Pigs housed in the CD chamber gained 27.2% more slowly (P less than .001; 75 vs 1.03 kg/d) than those in the TN environment and consumed 5.7% more feed (P less than .05; 3.88 vs 3.67 kg/d). The lower ADG and higher feed intake (FI) exhibited by the CD pigs resulted in poorer (P less than .05) feed efficiency (F/G; 5.33 vs 3.73, respectively). A temperature x sex interaction occurred for ADG but not for FI or F/G. Twenty-four-hour shrink for the CD pigs was 16.4% less than for the TN pigs (3.72 vs 4.45%, respectively); however, commingling did not affect shrinkage.


Fetal antigens in nonneoplastic conditions.
During studies that showed the presence of fetal antigens on the surface of human malignant melanoma tumor cells, polyvalent antisera specific for human fetal tissues of varying ages were developed. These reagents demonstrated varying patterns of expression of fetal antigens at different ages in various tissues of the human fetus. The possibility that nonneoplastic adult cells showing either maturation arrest or excessive proliferation also might express fetal antigens led to studies of human bone marrow. Although normal bone marrow cells expressed low levels of fetal antigens, large amounts were seen on bone marrow cells of patients with anemias due to iron, B12, or folic acid deficiencies, as well as on those with leukemia. Moreover, normal adult tissues adapted to long-term culture also expressed fetal antigens. After 3 weeks in organ culture adult human skin showed morphological changes similar to those seen in fetal periderm and strongly expressed fetal antigens. In addition, lymphoblasts in long-term cultured human lymphoid cell lines established from normal donors also carried surface fetal antigens. These latter antigens were shared with neoplastic B-cells (chronic lymphocytic leukemia) but not with T-cells. Their expression varied with the cell cycle. The reexpression of fetal antigens on malignant cells is thought to signal a basic derangement in the control of differentiation which is considered to be peculiar to neoplasia. However, these studies indicate that normal adult cells also may reexpress fetal antigens under circumstances unrelated to neoplasia but associated with either maturation arrest or rapid and excessive proliferation.


The Impact of Exercise during Radiation Therapy for Prostate Cancer on Fatigue and Quality of Life: A Systematic Review and Meta-analysis.
Radiation therapy (RT) is clinically proven to improve survival in men with prostate cancer. Despite these advantages, it is known to cause adverse effects such as fatigue. This review proposes to summarize the totality of evidence from randomized controlled trials regarding the effectiveness of exercise on fatigue in men with prostate cancer as a primary outcome. Quality of life was a secondary outcome. RCTs that explored the effect of exercise during RT on fatigue for men with prostate cancer were searched using MEDLINE, Embase, CINAHL, Cochrane Library, AMED, ClinicalTrials.gov, and ISRTCN registry. Reference lists of included studies and reviews were also examined. Trials were excluded if they included a mixed cohort of patients where data could not be extracted for prostate cancer patients alone or if the intervention took place after RT had finished. Preferred Reporting Items for Systematic Reviews and Meta-Analyses standardized reporting guidelines were followed to ensure the standardised conduct and reporting of the research. The search strategy yielded a total of 278 studies, of which five met the inclusion criteria. A meta-analysis pooled data of 392 participants using the Cochrane Review Manager 5.3 random-effects model (DerSimonian-Laird approach) with the post-test means of the control and intervention groups and associated standard deviations. Exercise was significant at alleviating fatigue when compared to the control group (standardized mean differences, -1.03; 95% confidence interval, -1.82 to -0.24). Exercise during RT is an effective approach to alleviate fatigue in men with prostate cancer. The effect on quality of life was not significant, possibly because of considerable heterogeneity across studies.


Should we adopt a prognosis-based approach to unexplained infertility?
The treatment of unexplained infertility is a contentious topic that continues to attract a great deal of interest amongst clinicians, patients and policy makers. The inability to identify an underlying pathology makes it difficult to devise effective treatments for this condition. Couples with unexplained infertility can conceive on their own and any proposed intervention needs to offer a better chance of having a baby. Over the years, several prognostic and prediction models based on routinely collected clinical data have been developed, but these are not widely used by clinicians and patients. In this opinion paper, we propose a prognosis-based approach such that a decision to access treatment is based on the estimated chances of natural and treatment-related conception, which, in the same couple, can change over time. This approach avoids treating all couples as a homogeneous group and minimizes unnecessary treatment whilst ensuring access to those who need it early.