Genetically encoded betaxanthin-based small-molecular fluorescent reporter for mammalian cells.
We designed and engineered a dye production cassette encoding a heterologous pathway, including human tyrosine hydroxylase and Amanita muscaria 4,5-DOPA dioxygenase, for the biosynthesis of the betaxanthin family of plant and fungal pigments in mammalian cells. The system does not impair cell viability, and can be used as a non-protein reporter system to directly visualize the dynamics of gene expression by profiling absorbance or fluorescence in the supernatant of cell cultures, as well as for fluorescence labeling of individual cells. Pigment profiling can also be multiplexed with reporter proteins such as mCherry or the human model glycoprotein SEAP (secreted alkaline phosphatase). Furthermore, absorbance measurement with a smartphone camera using standard application software enables inexpensive, low-tech reporter quantification.


Atypical diffusion tensor hemispheric asymmetry in autism.
Biological measurements that distinguish individuals with autism from typically developing individuals and those with other developmental and neuropsychiatric disorders must demonstrate very high performance to have clinical value as potential imaging biomarkers. We hypothesized that further study of white matter microstructure (WMM) in the superior temporal gyrus (STG) and temporal stem (TS), two brain regions in the temporal lobe containing circuitry central to language, emotion, and social cognition, would identify a useful combination of classification features and further understand autism neuropathology. WMM measurements from the STG and TS were examined from 30 high-functioning males satisfying full criteria for idiopathic autism aged 7-28 years and 30 matched controls and a replication sample of 12 males with idiopathic autism and 7 matched controls who participated in a previous case-control diffusion tensor imaging (DTI) study. Language functioning, adaptive functioning, and psychotropic medication usage were also examined. In the STG, we find reversed hemispheric asymmetry of two separable measures of directional diffusion coherence, tensor skewness, and fractional anisotropy. In autism, tensor skewness is greater on the right and fractional anisotropy is decreased on the left. We also find increased diffusion parallel to white matter fibers bilaterally. In the right not left TS, we find increased omnidirectional, parallel, and perpendicular diffusion. These six multivariate measurements possess very high ability to discriminate individuals with autism from individuals without autism with 94% sensitivity, 90% specificity, and 92% accuracy in our original and replication samples. We also report a near-significant association between the classifier and a quantitative trait index of autism and significant correlations between two classifier components and measures of language, IQ, and adaptive functioning in autism.


Biochemical characterization of MIP-1 alpha nuclear protein.
A family of hematopoietic specific transcription factors, MIP-1 alpha nuclear protein (MNP) family, has recently been identified. They are intimately involved in regulating the transcription of the huMIP-1 alpha gene in monocytes, T-cells, and transformed B-cells. One member of the family (MNP-1) is essential for promoter activity in monocytes and B-cells, while another (MNP-2) is required for full promotor activity in T-cells. A third member of the family (MNP-3) is expressed in PMA induced HL60 cells and probably has a role in monocyte differentiation. In this communication we demonstrate by two techniques that MNP-1 and MNP-2 are distinct but related factors, and we present further evidence to show that MNP-1 acts as a heterodimer.


Functional activation of glutamate ionotropic receptors in the developing mouse retina.
Ionotropic glutamate receptors have been associated with early development of the visual process by regulating cell differentiation, cell motility, and synaptic contacts. We determined the expression of functional ionotropic glutamate receptors during development of the mouse retina by assessing 1-amino-4-guanidobutane (agmatine; AGB) immunolabelling after application of a range of glutamate analogs. Colocalization of AGB with calretinin and islet-1 allowed the identification of functional receptors in neurochemically defined neurons. Activation with kainate (KA), alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA), and N-methyl-D-aspartate (NMDA) resulted in AGB entry into neurons consistent with that found previous receptor subunit localization studies in the developing retina. Temporal analysis revealed that application of 50 microM KA activated receptors as early as embryonic day 18 in the ventricular zone and in the ganglion cell layer, whereas 30 muM AMPA activated cells predominantly in the ganglion cell layer. Cholinergic amacrine cells showed functional KA and AMPA receptors upon their insertion into the conventional amacrine cell layer from postnatal day 1 (P1). OFF cone bipolar cells showed functional KA receptors from P6, at a developmental age when they are known to make contact with ganglion cells. NMDA activation led to diffuse AGB labeling at birth among cells in the ganglion cell layer, whereas, at P1, regularly spaced cholinergic amacrine cells in the conventional amacrine cell layer started to be responsive to NMDA. Non-NMDA receptors were first to show functional activation in the developing retina, and cholinergic amacrine cells displayed functional ionotropic glutamate receptors after reaching their final destination.


Stability and electrostatic assembly of au nanorods for use in biological assays.
The structure, stability, and aggregation potential of short Au nanorods under biological-based solution conditions have been studied. These attributes were studied using UV-vis spectroscopy, transmission electron microscopy, zeta-potential analysis, and dynamic light scattering. The stability and aggregation potential of the materials depended strongly upon both the purity and the solvent used to prepare Au nanorod solutions. When the Au nanorods were dissolved in Tris buffer at concentrations less than 10.0 mM, no aggregation was observed; however, when the solvent was comprised of Tris buffer with concentrations between 10.0 and 100 mM, significant aggregation of the materials occurred. This effect resulted in a dramatic broadening and shift in the absorbance maxima of the longitudinal surface plasmon resonance. At Tris buffer concentrations of greater than 100 mM, minimal to no aggregation of the materials in solution was observed. Such an ability is based upon electrostatic aggregation of the materials in solution mediated by the anions associated with the buffer system; at concentrations between 10.0 and 100 mM, the anions present electrostatically bind to the surfaces of the Au nanorods that are positively charged, resulting in cross-linking of the materials. At higher buffer concentrations, a sufficient number of anions are present in solution to template around the entire surface of each individual nanorod, in effect neutralizing the charge and producing an electronic double layer, which prevents aggregation. Such studies are timely as they represent an analysis of the stability and range of use of Au nanorods for biological-based applications where remarkable potential exists.


A genetic analysis of opioid-induced hyperalgesia in mice.
Opioid-induced hyperalgesia (OIH) is a syndrome of increased sensitivity to noxious stimuli, seen after both the acute and chronic administration of opioids, that has been observed in humans and rodent models. This syndrome may reduce the clinical utility of opioids in treating acute and chronic pain. In these studies, the authors measured the propensity of 15 strains of inbred mice to develop mechanical manifestations of OIH. These data were subjected to in silico genetic analysis, which resulted in the association of haplotypic blocks within or near several known genes. Both pharmacologic agents and transgenic mice were used to confirm the functional association of the most strongly linked gene with OIH. Both baseline mechanical nociceptive thresholds and the percentage changes in these thresholds after 4 days of morphine treatment were found to be highly strain dependent. The haplotypic blocks most strongly associated with the mechanical OIH data were located within the beta2 adrenergic receptor gene (beta2-AR). Using the selective beta2-AR antagonist butoxamine, the authors observed a dose-dependent reversal of OIH. Furthermore, deletion of the beta2-AR gene sharply reduced the mechanical allodynia present after morphine treatment in the wild-type mouse strain. Analysis of the associated beta2-AR haplotypic block identified single nucleotide polymorphisms potentially explaining in part the strain specific differences in OIH. Genetic variants of the beta2-AR gene seem to explain some part of the differences between various strains of mice to develop OIH. The association of this gene with OIH suggests specific pharmacologic strategies for reducing the impact of OIH on patients consuming opioids.


[Vascular endothelium (review). I. General morphology. 2B: phylogenesis of the vascular endothelium].
The phylogenetic descent of vascular endothelium from mesenchyme--derived precursors is described related to the development of a vessel--bound microcirculation. Endothelial precursors in primitive animals may have migrated into tissue clefts gradually forming vascular tubes. True microcirculatory vessels at first appear in the nemertines, a closed vascular system is present in some annelids whereas in arthropods an open lacunar system predominates. The first appearance of true endotheliocytes is under discussion; the author gives some evidence that it is present already in some annelids. Precursor of the endothelial wall of vessels may be the so called "Leydig's membrane", covered with amoebocytes and other mesenchymal cells. The molluscs exhibit many variants of endothelium. In the fishes, the vascular system begins to split into a blood and a lymphatic system. Obviously the specialization of endothelium correlates with the level of evolution. Despite the complicated course, the evolution of endothelium may be regarded as monophyletic.


Solution conformation of the (+)-trans-anti-[BP]dG adduct opposite a deletion site in a DNA duplex: intercalation of the covalently attached benzo[a]pyrene into the helix with base displacement of the modified deoxyguanosine into the major groove.
This paper reports on the solution structure of the (+)-trans-anti-[BP]dG adduct positioned opposite a deletion site in a DNA oligomer duplex which defines the alignment of this covalent benzo[a]pyrene-N2-deoxyguanosine stereosiomer relative to the deletion site. The combined NMR-molecular mechanics computation studies were undertaken on the (+)-trans-anti-[BP]dG adduct embedded in the d(C5-[BP]G6-C7).d(G16-G17) sequence context in a duplex containing 11 residues on the modified strand and 10 on the partner, with no base opposite the modification. The exchangeable and nonexchangeable protons of the benzo[a]pyrenyl moiety and the nucleic acid were assigned following analysis of two-dimensional NMR data sets in H2O and D2O solution. The solution conformation of the (+)-trans-anti-[BP]dG.del 11-mer duplex has been determined by incorporating intramolecular and intermolecular proton-proton distances defined by lower and upper bounds deduced from NOESY spectra as restraints in molecular mechanics computations in torsion angle space. The benzo[a]pyrene ring of [BP]dG6 is intercalated between intact Watson-Crick dC5.dG17 and dC7.dG16 base pairs with the deoxyguanosine base of [BP]dG6 displaced into the major groove. The intercalation site is wedge shaped, being narrower toward the dG16-dG17 step on the deletion-containing strand. The deoxyguanosine base of [BP]dG6 which is positioned in the major groove is inclined relative to the helix axis and stacks over the 5'-flanking dC5 residue in the solution structure. The intercalative-base displacement structure of the (+)-trans-anti-[BP]dG.del 11-mer duplex exhibits several unusually shifted proton resonances which can be readily accounted for by the ring current contribution of the deoxyguanosyl and pyrenyl rings of the [BP]dG6 adduct. This solution structure of the (+)-trans-anti-[BP]dG.del 11-mer duplex where the pyrene ring intercalates into the helix with displacement of the modified deoxyguanosine into the major groove strikingly contrasts with our previous study on the (+)-trans-anti-[BP]dG.dC 11-mer duplex [Cosman, M., et al. (1992) Proc. Natl. Acad. Sci. U.S.A. 89, 1914-1918] where the benzo[a]pyrene ring is positioned in the minor groove without disruption of the Watson-Crick pairing at the [BP]dG.dC modification site. Thus, generation of the deletion site following removal of the dC opposite the (+)-trans-anti-[BP]dG results in a displacement of the entire [BP]dG residue toward the major groove and intercalation of the benzo[a]pyrene ring into the helix.


Scapular Muscle Activity During Static Yoga Postures.
Study Design Controlled, cross-sectional laboratory study. Background Despite the growing popularity of yoga, little is known about the muscle activity of the scapular stabilizers during isometric yoga postures and their potential utility in shoulder rehabilitation. Objectives To examine scapular stabilizer muscle activation during various yoga postures. Methods Twenty women with yoga experience and no shoulder pain or injury participated. Electromyography was used to record the muscle activity of the upper, middle, and lower trapezius, as well as of the serratus anterior, during 15 yoga postures. Results Muscle activity varied between yoga postures (3%-57% maximum voluntary isometric contraction [MVIC]). Overall, the "locust arms forward" posture elicited the highest activity from the upper (22.4% MVIC), middle (41.8% MVIC), and lower (56.8% MVIC) trapezius, while several postures elicited moderate activity (greater than 20% MVIC) from the serratus anterior. Conversely, the "dancer's pose right," "reverse tabletop," and "warrior II" postures demonstrated low activity (less than or equal to 15.7% MVIC) of the scapular stabilizers. Conclusion Strengthening the scapular stabilizer muscles is an important component of shoulder rehabilitation. Yoga postures have been identified that activate the scapular stabilizer muscles at varying levels of activity. J Orthop Sports Phys Ther 2018;48(6):504-509. Epub 6 Apr 2018. doi:10.2519/jospt.2018.7311.


Determinants of anti-PD-1 response and resistance in clear cell renal cell carcinoma.
ADAPTeR is a prospective, phase II study of nivolumab (anti-PD-1) in 15 treatment-naive patients (115 multiregion tumor samples) with metastatic clear cell renal cell carcinoma (ccRCC) aiming to understand the mechanism underpinning therapeutic response. Genomic analyses show no correlation between tumor molecular features and response, whereas ccRCC-specific human endogenous retrovirus expression indirectly correlates with clinical response. T cell receptor (TCR) analysis reveals a significantly higher number of expanded TCR clones pre-treatment in responders suggesting pre-existing immunity. Maintenance of highly similar clusters of TCRs post-treatment predict response, suggesting ongoing antigen engagement and survival of families of T cells likely recognizing the same antigens. In responders, nivolumab-bound CD8+ T cells are expanded and express GZMK/B. Our data suggest nivolumab drives both maintenance and replacement of previously expanded T cell clones, but only maintenance correlates with response. We hypothesize that maintenance and boosting of a pre-existing response is a key element of anti-PD-1 mode of action.


Genetic ancestry and ethnic identity in Ecuador.
We investigated the ancestral origins of four Ecuadorian ethnic groups-Afro-Ecuadorian, Mestizo, Montubio, and the Indigenous Tsáchila-in an effort to gain insight on the relationship between ancestry, culture, and the formation of ethnic identities in Latin America. The observed patterns of genetic ancestry are largely concordant with ethnic identities and historical records of conquest and colonization in Ecuador. Nevertheless, a number of exceptional findings highlight the complex relationship between genetic ancestry and ethnicity in Ecuador. Afro-Ecuadorians show far less African ancestry, and the highest levels of Native American ancestry, seen for any Afro-descendant population in the Americas. Mestizos in Ecuador show high levels of Native American ancestry, with substantially less European ancestry, despite the relatively low Indigenous population in the country. The recently recognized Montubio ethnic group is highly admixed, with substantial contributions from all three continental ancestries. The Tsáchila show two distinct ancestry subgroups, with most individuals showing almost exclusively Native American ancestry and a smaller group showing a Mestizo characteristic pattern. Considered together with historical data and sociological studies, our results indicate the extent to which ancestry and culture interact, often in unexpected ways, to shape ethnic identity in Ecuador.


Decapping activators in Saccharomyces cerevisiae act by multiple mechanisms.
Eukaryotic mRNA degradation often occurs in a process whereby translation initiation is inhibited and the mRNA is targeted for decapping. In yeast cells, Pat1, Scd6, Edc3, and Dhh1 all function to promote decapping by an unknown mechanism(s). We demonstrate that purified Scd6 and a region of Pat1 directly repress translation in vitro by limiting the formation of a stable 48S preinitiation complex. Moreover, while Pat1, Edc3, Dhh1, and Scd6 all bind the decapping enzyme, only Pat1 and Edc3 enhance its activity. We also identify numerous direct interactions between Pat1, Dcp1, Dcp2, Dhh1, Scd6, Edc3, Xrn1, and the Lsm1-7 complex. These observations identify three classes of decapping activators that function to directly repress translation initiation and/or stimulate Dcp1/2. Moreover, Pat1 is identified as critical in mRNA decay by first inhibiting translation initiation, then serving as a scaffold to recruit components of the decapping complex, and finally activating Dcp2.


Hyperglycemia Aggravates Diet-Induced Coronary Artery Disease and Myocardial Infarction in SR-B1-Knockout/ApoE-Hypomorphic Mice.
Diabetes is a risk factor for development of atherosclerotic cardiovascular disease. Animal model studies in mice revealed that hyperglycemia increases development of atherosclerosis in the aorta as well as myocardial fibrosis in surgical models of coronary artery ligation; however, the impact of hyperglycemia on coronary artery atherosclerosis and subsequent heart disease is less clear. To investigate the effect of hyperglycemia on atherosclerosis and coronary heart disease, we used a mouse model of diet-induced coronary artery atherosclerosis and myocardial infarction, the high fat/high cholesterol (HFC) diet fed SR-B1 knockout (KO)/apoE-hypomorphic (HypoE) mouse. Hyperglycemia was induced in these mice by streptozotocin (STZ) treatment. This increased HFC diet-dependent atherosclerosis development (p = 0.02) and necrotic core formation (p = 0.0008) in atherosclerotic plaques in the aortic sinus but did not increase the extent of atherosclerosis in coronary arteries. However, it did increase the extent of platelet accumulation in atherosclerotic coronary arteries (p = 0.017). This was accompanied by increased myocardial fibrosis (p = 0.005) and reduced survival (p = 0.01) compared to control-treated, normoglycemic mice. These results demonstrate that STZ-treatment exerted differential effects on the level of atherosclerosis in the aortic sinus and coronary arteries. These results also suggest that SR-B1-KO/HypoE mice may be a useful non-surgical model of diabetic cardiomyopathy in the context of coronary artery atherothrombosis.


Phenolato Ti(IV) hexacoordinate complexes for anticancer chemotherapy: enhancement of solubility, hydrolytic stability, and cytotoxicity.
A new series of five titanium(IV) complexes based on diaminobis(phenolato)-bis(alkoxo) ligands with different substitutions was synthesized and characterized. All complexes were analyzed by X-ray crystallography, and all structures indicated C2 symmetrical octahedral compounds. All complexes exhibited enhanced solubility in aqueous media compared with the parent methylated derivative phenolaTi (up to 0.4 vs. 0.05 mg ml-1 of phenolaTi) due to halogen and alkoxo/hydroxo substitutions, with particularly enhanced water solubility for the methoxylated and hydroxylated derivatives. In particular, high hydrolytic stability was recorded for all derivatives, with the t½ for ligand hydrolysis of more than 8 days, as established by 1H NMR and HR-MS. All complexes were cytotoxic toward human ovarian A2780, colon HT-29, and cervical HeLa cancer cell lines (IC50 values in the range of 0.3-40 μM), with negligible activity toward non-cancerous MRC-5 cells. The halogenated compounds of this series exhibit the best combination of stability and activity, making them highly promising for anticancer applications.


Protection by dimethylthiourea against retinal light damage in rats.
The protective effect of dimethylthiourea (DMTU) against retinal light damage was determined in albino rats reared in darkness or in weak cyclic light. Rats maintained under these conditions were treated with DMTU at different concentrations and dosing schedules and then exposed for various times to intense visible light, either intermittently (1 hr light and 2 hr dark) or continuously. The extent of retinal light damage was determined 2 weeks after light exposure by comparing rhodopsin levels in experimental rats with those in unexposed control animals. To determine the effect of DMTU on rod outer segment (ROS) membrane fatty acids, ROS were isolated immediately after intermittent light exposure, and fatty acid compositions were measured. The time course for DMTU uptake and its distribution in serum, retina, and the retinal pigment epithelium (RPE)/choroid complex was determined in other rats not exposed to intense light. After intraperitoneal injection of the drug (500 mg/kg body weight), DMTU appeared rapidly in the serum, retina, and the RPE and choroid. In the ocular tissues, it was distributed 70-80% in the retina and 20-30% in the RPE and choroid. This antioxidant appears to have a long half-life because it was present in these same tissues 72 hr after a second intraperitoneal injection. For rats reared in the weak cyclic light environment, DMTU (two injections) provided complete protection against rhodopsin loss after intense light exposures of up to 16 hr. Only 15% rhodopsin loss was found in cyclic-light DMTU-treated rats after 24 hr of intermittent or continuous light. For rats reared in darkness, DMTU treatment resulted in a rhodopsin loss of less than 20% after 8-16 hr of continuous light and approximately 40% after similar exposure to intermittent light. Irrespective of the type of light exposure, rhodopsin loss in the dark-reared DMTU-treated rats was nearly identical to that found in uninjected cyclic light-reared animals. In rats from both light-rearing environments, DMTU treatment prevented the light-induced loss of docosahexaenoic acid from ROS membranes. As measured by rhodopsin levels 2 weeks later, DMTU was most effective when given as two doses administered 24 hr before and just before intense light exposure. As a single dose given during continuous light exposure, DMTU protected cyclic light-reared rats for at least 4 hr after the start of exposure but was ineffective in dark-reared animals if injected 1 hr after the start of light. It was also ineffective in both types of rats when given after light exposure.(ABSTRACT TRUNCATED AT 400 WORDS)


The molecular basis for IL-31 production and IL-31-mediated itch transmission: from biology to drug development.
Atopic dermatitis (AD) is one of the most prevalent chronic inflammatory skin diseases in the world. It is characterized by recurrent eczematous lesions and intense itch, and many cytokines are involved in the pathogenesis of AD. Among them, much attention has been paid to interleukin 31 (IL-31) as an AD-associated itch mediator. IL-31 is mainly produced by CD4+ helper T cells and transmits the signals via a heterodimeric receptor composed of IL-31 receptor A (IL-31RA) and oncostatin M receptor (OSMR), both of which are expressed in dorsal root ganglion (DRG) neurons. However, the molecular mechanisms of how IL-31 is produced in helper T cells upon stimulation and transmits the itch sensation to the brain were largely unknown. Recently, by using original mouse models of AD, we have identified endothelial PAS domain 1 (EPAS1) and neurokinin B (NKB) as key molecules critical for IL-31 production and IL-31-mediated itch transmission, respectively. These molecules could be novel drug targets for AD-associated itch. This review highlights our recent findings, which show the functional significance of these molecules in the IL-31-induced itch sensation, referring to their application to drug development.


Excitatory amino acids in cerebrospinal fluid of patients with acute head injuries.
The excitatory amino acids (EAAs) glutamate (Glu) and aspartate (Asp) play a role in the pathogenesis of postischemic and posttraumatic brain insult. The changes of EAAs in cerebrospinal fluid (CSF) of patients with traumatic brain injury are incompletely understood. We used reversed-phase HPLC with precolumn derivatization with o-phthalaldehyde and fluorescence detection to measure Glu and Asp in CSF of 42 patients with acute head injury and 9 control adults without neurologic diseases. We assessed the Glasgow Coma Scale (GCS) on admission, the main lesion patterns on computed tomography (CT) scan within 24 h post trauma, and the Glasgow Outcome Scale (GOS) 3 months post injury. The mean concentrations of Glu and Asp in CSF in the brain-injured group were significantly higher than those of the control group (Glu, P <0.001; Asp, P <0.05). In patients admitted within 24 h after severe injury (n = 13), peak Glu values appeared within 48 h in 11 patients (85%), and the mean value remained higher than control values at day 7 (P <0.02). The concentrations of EAAs were higher in patients with severe injuries (GCS < or =8) than in those with milder injuries (Glu, P <0.001; Asp, P <0.05). GCS and peak EAAs correlated negatively (Glu, rs = -0.5706, P <0.001; Asp, rs = -0.5503, P <0.001). The patients with focal brain contusion on initial CT scan (n = 8) had a significantly lower peak Glu value than the patients with other patterns (n = 8-15; P <0.02 to 0.001). The peak value of EAAs in the poor-outcome group (including severe disability, vegetative state, and death) was significantly higher than in the good-outcome group (good recovery and moderate disability; Glu, P <0.001; Asp, P <0.01); GOS was closely correlated to the EAA values (Glu, rs = 0.5737, P <0.001; Asp, rs = 0.5470, P <0.001). The EAA concentrations in CSF increase after acute head injury and remain higher for at least 1 week post injury in severely injured patients. The more severe the trauma, the more obvious the excitotoxicity induced by EAAs and the worse the outcome.


Reconstruction of Large Acquired Scalp Defects: Ten-Year Experience.
The method of choice for reconstruction of large acquired scalp defects depends on numerous factors. The aim of our study was to analyze reconstructions of large acquired scalp defects performed on 135 patients in the period of 10 years. We have monitored the factors with an impact on the reconstruction method, complications, and the achieved results. Skin grafts, local, regional and free flaps, have been applied for reconstruction of defects. The depth, size, defect localization, condition of surrounding tissue, co-morbidities and causes of occurrence of defect have an impact on the method of choice for defect reconstruction. Acquired scalp defects are the most frequent in the older population and, in most cases, occur upon the surgical removal of malignant tumors. Local fasciocutaneous flaps are the method of choice for small and acquired scalp defects of medium size while free flaps are the best solution for reconstruction of the large full-thickness scalp defects. Frequency of severe complications is significantly higher in the patients who undergo craniotomy, or those with liquorrhea.


Development of an Evidence-Based Employee Wellness Program.
Workplaces can positively or negatively shape employee health and sense of well-being. Employees who feel well are more productive, engaged, and present at work. The effects of the COVID-19 pandemic added to an already stressed work environment for educators. Student success is dependent on educators who are healthy and present. Employee wellness programs have the potential to improve the well-being of employees and now, more than ever, should be a priority in schools. The purpose of this article is to provide the school nurse with evidence-based resources to assist in the creation and implementation of an employee wellness program in the school setting.


Factitious hyperamylasuria by a nurse: symptom of Munchausen's syndrome.
A 30-year-old nurse presented with abdominal pain and tenderness. Her blood tests, including amylase, were normal. Urinary amylase was extremely high. The source of the increased urinary amylase was found to be the patient's saliva - she had spit into her urine sample. Subsequent investigation showed that she had Munchausen's syndrome.


X-ray evidence for metal-N-7 bonding in a hydrated manganese derivative of guanosine 5'-monophosphate.
Single-crystal X-ray studies of a manganese(II) derivative of guanosine 5'-monophosphate, [Mn(5'-GMP)(H(2)O)(5)],3H(2)O, have shown that it is isostructural with its nickel analogue. The manganese atom therefore is bonded to five water molecules with the remaining octahedral co-ordination site being occupied by N-7 of the nucleotide base. No direct metal-phosphate bonding is involved, but there are structure-stabilizing intramolecular hydrogen bonds between two phosphate oxygen atoms and co-ordinated water molecules.


[Changes in glutamate release in the rat nucleus accumbens during food and pain reinforcement].
In vivo microdialysis combined with HPLC-EC analysis was used to monitor extracellular glutamate in the n. accumbens of Sprague-Dawley rats during footshock and food delivery. The footshock presentation resulted in a delayed increase in extracellular glutamate level, whereas the food intake caused its decrease. The intra-accumbens infusion of glutamate reuptake blocker D,L-threo-beta-hydroxiaspartate (1 mM) completely prevented the food-induced decrease in glutamate level. The intra-accumbens infusion of sodium channel blocker tetrodotoxin (1 microM) led to an increase in glutamate extracellular level in the n. accumbens in response to food intake. The results suggest that the food-induced decrease in glutamate extracellular level in the n. accumbens occurs due to an enhancement of high-affinity glutamate uptake that is probably under the neuronal control during feeding.


The Chromatin Remodeling Component Arid1a Is a Suppressor of Spontaneous Mammary Tumors in Mice.
Human cancer genome studies have identified the SWI/SNF chromatin remodeling complex member ARID1A as one of the most frequently altered genes in several tumor types. Its role as an ovarian tumor suppressor has been supported in compound knockout mice. Here, we provide genetic and functional evidence that Arid1a is a bona fide mammary tumor suppressor, using the Chromosome aberrations occurring spontaneously 3 (Chaos3) mouse model of sporadic breast cancer. About 70% of mammary tumors that formed in these mice contained a spontaneous deletion removing all or part of one Arid1a allele. Restoration of Arid1a expression in a Chaos3 mammary tumor line with low Arid1a levels greatly impaired its ability to form tumors following injection into cleared mammary glands, indicating that ARID1A insufficiency is crucial for maintenance of these Trp53-proficient tumors. Transcriptome analysis of tumor cells before and after reintroduction of Arid1a expression revealed alterations in growth signaling and cell-cycle checkpoint pathways, in particular the activation of the TRP53 pathway. Consistent with the latter, Arid1a reexpression in tumor cells led to increased p21 (Cdkn1a) expression and dramatic accumulation of cells in G2 phase of the cell cycle. These results not only provide in vivo evidence for a tumor suppressive and/or maintenance role in breast cancer, but also indicate a potential opportunity for therapeutic intervention in ARID1A-deficient human breast cancer subtypes that retain one intact copy of the gene and also maintain wild-type TRP53 activity.


Acid-fastness of fungi in blastomycosis and histoplasmosis.
In 60% of the infections caused by Blastomyces dermatitidis and in 47% of the infections caused by Histoplasma capsulatum, some of the organisms are acid-fast in tissue sections. Treatment with 1N hydrochloric acid at 60 degrees C abolishes the acid-fastness of H capsulatum, which indicates that the acid-fastness may be related to mycolic acid. Organisms of B dermatitidis, however, remain acid-fast in 83% of the sections treated with acid. The presence of acid-fastness in small cells can aid in the differentiation of B dermatitidis and H capsulatum from Cryptococcus neoformans and Candida albicans, Persistent acid-fastness of organisms after treatment with heated acid helps identify B dermatitidis in tissue sections, especially when the strain is small or in other ways atypical.


New imaging tools for the diagnosis of infection.
Infection imaging became widespread in 1971 with the release of 67Ga citrate. Multiphase skeletal scintigraphy and radiolabeled white blood cells (WBCs) have since become the most widespread clinically used agents for the imaging of infection. A wide variety of other radiolabeled probes are under investigation, based on antibodies, cytokines, assorted proteins and other molecules, alone or in various combinations. However, these latter agents, with a few exceptions, are not routinely used clinically. Radiolabeled ciprofloxacin represents the first attempt to develop an infection-specific imaging agent (most infection-imaging probes localized nonspecifically to inflammation as well), but it has not proven superior to radiolabeled WBCs or 18F-fluoro-deoxy-glucose (FDG) PET. Because of the ability to combine exquisite anatomic detail with focal uptake of 18F-FDG, PET-computed tomography has achieved great success in the detection and localization of infection, including in clinically adverse conditions. Despite these advances, at this time an infection-specific imaging agent does not exist.


Neural Mechanisms of Social and Nonsocial Reward Prediction Errors in Adolescents with Autism Spectrum Disorder.
Autism spectrum disorder (ASD) is characterized by impaired predictive abilities; however, the neural mechanisms subsuming reward prediction errors in ASD are poorly understood. In the current study, we investigated neural responses during social and nonsocial reward prediction errors in 22 adolescents with ASD (ages 12-17) and 20 typically developing control adolescents (ages 12-18). Participants performed a reward prediction error task using both social (i.e., faces) and nonsocial (i.e., objects) rewards during a functional magnetic resonance imaging scan. Reward prediction errors were defined in two ways: (a) the signed prediction error, the difference between the experienced and expected reward; and (b) the thresholded unsigned prediction error, the difference between expected and unexpected outcomes regardless of magnitude. During social reward prediction errors, the ASD group demonstrated the following differences relative to the TD group: (a) signed prediction error: decreased activation in the right precentral gyrus and increased activation in the right frontal pole; and (b) thresholded unsigned prediction error: increased activation in the right anterior cingulate gyrus and bilateral precentral gyrus. Groups did not differ in brain activation during nonsocial reward prediction errors. Within the ASD group, exploratory analyses revealed that reaction times and social-communication impairments were related to precentral gyrus activation during social prediction errors. These findings elucidate the neural mechanisms of social reward prediction errors in ASD and suggest that ASD is characterized by greater neural atypicalities during social, relative to nonsocial, reward prediction errors in ASD. Autism Res 2020, 13: 715-728. © 2020 International Society for Autism Research, Wiley Periodicals, Inc. LAY SUMMARY: We used brain imaging to evaluate differences in brain activation in adolescents with autism while they performed tasks that involved learning about social and nonsocial information. We found no differences in brain responses during the nonsocial condition, but differences during the social condition of the learning task. This study provides evidence that autism may involve different patterns of brain activation when learning about social information.


Oxidation potentials of alpha-hydroxyalkyl radicals in acetonitrile obtained by photomodulated voltammetry.
Oxidation potentials, E1/2(ox) of alpha-hydroxyalkyl radicals of the type R(1)R(2)C(*)OH (denoted K(1)H(*)) have been obtained in acetonitrile by photomodulated voltammetry. The values of E1/2(ox) increase as the R(1) and R(2) groups are changed from alkyl to aryl and, in particular, strong electron-withdrawing functionalities such as CN and CF3. Using rate data available in the literature for the pinacol photoexchange reaction K + K(1)H(*) --> KH(*) + K(1), it is found that as the difference in the standard potential of the ketone K, EK degrees and the oxidation potential of K(1)H(*), E1/2(ox), increases there is a modest increase in the exchange rate constant, k(ex). This indicates that even if some charge transfer may occur between the hydroxyalkyl radical and the ketone in the transition state, it is certainly not to the extent of a complete electron transfer. If the exchange reaction is treated as a simple hydrogen atom transfer process within the Marcus model, the intrinsic barrier is found to be 8-13 kcal mol(-1) due to the changes occurring in bonds, hybridizations, and bond angles. Finally, acid dissociation constants for K(1)H(*) are provided by means of a thermochemical cycle.


Fasting blood glucose level and prognosis in non-small cell lung cancer (NSCLC) patients.
Diabetes has been consistently linked to many forms of cancers, such as liver, colorectal, pancreatic, and breast cancer, however, the role of diabetes in outcome among cancer patients remains unclear. In this study, we retrospectively reviewed electronic medical records of 342 inpatients newly diagnosed with NSCLC referred by a teaching hospital cancer center in southern Taiwan between 2005 and 2007 to examine the effects of fasting glucose levels at time of cancer diagnosis on overall survival in patients with non-small cell lung cancer (NSCLC). All patients were followed up until the end of 2010. The Kaplan-Meier method was used to compare survival curves for patients with and without diabetes. The Cox proportional hazards model was used to estimate hazard ratios for the association between diabetes, other prognostic factors and patient survival. We observed that significant prognostic factors for poor overall survival in patients with NSCLC included older age, smoking, poor performance status, advanced stage (stage IIIB or IV), and no cancer-directed surgery treatment. Particularly, we identified that diabetic state defined by fasting blood glucose level ≥126 mg/dl was another independent prognostic factor for these patients. Compared with those who had normal range of fasting glucose level (70-99 mg/dl), patients with high fasting glucose level (≥126 mg/dl) had 69% excess risk of all-cause mortality in patients with NSCLC. Diabetes as indicated by elevated fasting blood glucose was independently associated with a significantly higher risk of all-cause mortality in patients with NSCLC, indicating that diabetes or hyperglycemia effectively controlled may present an opportunity for improving prognosis in NSCLS patients with abnormal glucose level.


Inducible expression of murine IP-10 mRNA varies with the state of macrophage inflammatory activity.
We have examined the expression of inducible inflammatory genes in murine macrophages from different tissues and at different stages of inflammatory activity. Although i.v. administration of IFN-gamma (10,000 U/mouse) strongly induced expression of IP-10 mRNA in the adherent cell population of the spleen, thioglycollate-elicited peritoneal macrophages were essentially unresponsive at the same dose. In contrast, D3 mRNA was expressed in both cell populations. This differential sensitivity of IP-10 mRNA expression was not restricted to stimulation by IFN-gamma as it was also seen when LPS (25 micrograms/mouse) was administered i.v. Expression of JE and KC mRNA, which encode cytokines related to IP-10, were also differentially expressed in elicited peritoneal macrophages from mice injected with LPS. Differential sensitivity was at least partially related to the state of macrophage activation because IP-10 mRNA was highly inducible in resident but not thioglycollate-elicited peritoneal macrophages. The eliciting agent was also an important determinant because proteose-peptone-elicited peritoneal macrophages were nearly as sensitive as splenic macrophages with respect to expression of IP-10 mRNA. IFN-gamma treatment induced IP-10 and D3 mRNA rapidly and transiently with the same time course in the spleen. IP-10 mRNA was not induced by IFN-gamma in TG-elicited macrophages regardless of the time after treatment. This differential expression of IP-10 was a consequence of different concentration requirements for IFN-gamma in the two cell types; thioglycollate-elicited macrophages required five- to 10-fold more IFN-gamma than did resident cells to achieve comparable IP-10 mRNA levels whether the agent was provided in vitro or in vivo. Thus variable sensitivity for induction of IP-10 mRNA was a characteristic of the macrophage itself and was not mediated by other cellular or molecular elements present in the inflammatory peritoneal cavity. The reduced sensitivity to IFN-gamma or LPS for expression of IP-10, JE, and KC mRNA as compared with TNF-alpha or D3 mRNA suggests that this distinct pattern of regulation may be restricted to members of these two related cytokine gene families that exhibit cell-type specific chemoattractant activity.


An adaptive view of attentional control.
Although humans can voluntarily direct their attention to particular stimuli, attention can at times be involuntarily allocated to stimuli and such attentional capture can result in unproductive distraction. A challenge to any comprehensive theory of attention is to explain how involuntary mechanisms of attentional control and their potential to produce distraction are ultimately reflective of an adaptation. Traditional arguments on this topic have appealed to a generalized cost-benefit accounting. Specifically, the cost of misallocating attention to the kinds of stimuli prioritized by involuntary mechanisms of attentional control over the long run is argued to be small in comparison with the potentially life-altering cost of failing to attend to such stimuli, which involuntary mechanisms of attentional control guard against. Our understanding of these mechanisms has undergone a revolution in recent years, findings from which point to a much more sophisticated adaptation that systematically maximizes benefits associated with automating the control of attention while minimizing unwanted distraction. In this review, I provide an updated model of the adaptive nature of involuntary mechanisms of attentional control, outlining concrete principles governing the management of specific costs and benefits. I conclude that distraction does not in general constitute a failure of attentional control but rather reflects the joint product of these adaptive principles. (PsycInfo Database Record (c) 2022 APA, all rights reserved).


The N2 event-related potential reflects attention deficit in schizophrenia.
Schizophrenia involves deficits in detection and filtering of irrelevant stimuli. The N2 event-related potential (ERP), most likely reflecting classification of deviant stimuli, remains largely unstudied in this group. Recently, we reported that N2 amplitude correlated highly with reduced neocortical and medial temporal lobe volumes in schizophrenics. However, little is known about the functional properties of N2 in schizophrenics. To that end, the latency and amplitude of N2 were assessed in schizophrenic and control subjects to differently pitched tones. Subjects pressed a button in response to low probability (p = .15) target tones interspersed among high probability standard tones. Tones were either NEAR (1.4 kHz standard, 1.5 kHz target) or FAR (1 kHz standard, 1.5 kHz target) in pitch. N2 was measured from difference waveforms, subtracting ERPs on a simple reaction time task (target p = 1.0) from those of the detection task. Schizophrenics performed the detection task nearly as well as controls in the FAR condition, and more poorly in the NEAR condition. Schizophrenics displayed virtually no N2 amplitudes in either condition. The results are interpreted as electrophysiological signs of disturbance in stimulus classification and attention processes in schizophrenia directly related to pathology of N2 neural generators, independent of sensory or detectability problems.


UV Chromosomes and Haploid Sexual Systems.
The evolution of sex determination continues to pose major questions in biology. Sex-determination mechanisms control reproductive cell differentiation and development of sexual characteristics in all organisms, from algae to animals and plants. While the underlying processes defining sex (meiosis and recombination) are conserved, sex-determination mechanisms are highly labile. In particular, a flow of new discoveries has highlighted several fascinating features of the previously understudied haploid UV sex determination and related mating systems found in diverse photosynthetic taxa including green algae, bryophytes, and brown algae. Analyses integrating information from these systems and contrasting them with classical XY and ZW systems are providing exciting insights into both the universality and the diversity of sex-determining chromosomes across eukaryotes.


Bariatric surgery to treat type 2 diabetes: what is the recent evidence?
To critically discuss the strengths and shortcomings of recent progress achieved with bariatric surgery in patients with type 2 diabetes mellitus (T2DM), focussing on efficacy aspects (remission of diabetes and cardiovascular comorbidities). Despite an increasing armamentarium of pharmacotherapeutics to overcome several challenges, only 10% of T2DM patients achieve a composite goal of HbA1c, blood pressure and lipids. Bariatric surgery has emerged as a solution to these challenges in morbid obesity. Whether the same advantages can be translated into T2DM remains a matter of debate, certainly regarding safety, durability of diabetes recovery and long-term outcome. Bariatric surgery in T2DM patients with a BMI of at least 35 kg/m(2) has been shown to result in a 56% excess body weight loss, resolution of hypertension in 62%, amelioration of dyslipidaemia in greater than 70% and diabetes remission in 57-95%, depending on the type of surgery and the definition of diabetes resolution. These impressive results, and the fact that diabetes recovery often occurs before prominent weight loss is evident, have urged bariatric surgeons to consider surgical procedures as a valuable approach for diabetes control and diabetes remission in patients with a BMI ranging between 30 and 35 kg/m(2). Bariatric surgery is emerging as a valid option to treat T2DM, improving glycaemia and cardiovascular risk factors. However, there needs to be an agreed definition of resolution of diabetes in future studies and long-term efficacy is to be proven. For now, the challenge is to determine how to offer bariatric surgery in a responsible fashion.


In diabetic male Wistar rats, quercetin-conjugated superparamagnetic iron oxide nanoparticles have an effect on the SIRT1/p66Shc-mediated pathway related to cognitive impairment.
Quercetin (QC) possesses a variety of health-promoting effects in pure and in conjugation with nanoparticles. Since the mRNA-SIRT1/p66Shc pathway and microRNAs (miRNAs) are implicated in the oxidative process, we aimed to compare the effects of QC and QC-conjugated superparamagnetic iron oxide nanoparticles (QCSPIONs) on this pathway. Through the use of the chemical coprecipitation technique (CPT), SPIONs were synthesized, coated with dextran, and conjugated with quercetin. Adult male Wistar rats were given intraperitoneal injections of streptozotocin to look for signs of type 1 diabetes (T1D). The animals were randomized into five groups: the control group got deionized water (DI), free QC solution (25 mg/kg), SPIONs (25 mg/kg), and QCSPIONs (25 mg/kg), and all groups received repeat doses administered orally over 35 days. Real-time quantitative PCR was used to assess the levels of miR-34a, let-7a-p5, SIRT1, p66Shc, CASP3, and PARP1 expression in the hippocampus of diabetic rats. In silico investigations identified p66Shc, CASP3, and PARP1 as targets of let-7a-5p and miR-34a as possible regulators of SIRT1 genes. The outcomes demonstrated that diabetes elevated miR-34a, p66Shc, CASP3, and PARP1 and downregulated let-7a-5p and SIRT1 expression. In contrast to the diabetic group, QCSPIONs boosted let-7a-5p expression levels and consequently lowered p66Shc, CASP3, and PARP1 expression levels. QCSPIONs also reduced miR-34a expression, which led to an upsurge in SIRT1 expression. Our results suggest that QCSPIONs can regulate the SIRT1/p66Shc-mediated signaling pathway and can be considered a promising candidate for ameliorating the complications of diabetes.


Smad1 and 5 but not Smad8 establish stem cell quiescence which is critical to transform the premature hair follicle during morphogenesis toward the postnatal state.
Hair follicles (HFs) are regenerative miniorgans that offer a highly informative model system to study the regulatory mechanisms of hair follicle stem cells (hfSCs) homeostasis and differentiation. Bone morphogenetic protein (BMP) signaling is key in both of these processes, governing hfSCs quiescence in the bulge and differentiation of matrix progenitors. However, whether canonical or noncanonical pathways of BMP signaling are responsible for these processes remains unresolved. Here, we conditionally ablated two canonical effectors of BMP signaling, Smad1 and Smad5 during hair morphogenesis and postnatal cycling in mouse skin. Deletion of Smad1 and Smad5 (dKO) in the epidermis during morphogenesis resulted in neonatal lethality with lack of visible whiskers. Interestingly, distinct patterns of phospho-Smads (pSmads) activation were detected with pSmad8 restricted to epidermis and pSmad1 and pSmad5 exclusively activated in HFs. Engraftment of dKO skin revealed retarded hair morphogenesis and failure to differentiate into visible hair. The formation of the prebulge and bulge reservoir for quiescent hfSCs was precluded in dKO HFs which remained in prolonged anagen. Surprisingly, in postnatal telogen HFs, pSmad8 expression was no longer limited to epidermis and was also present in dKO bulge hfSCs and matrix progenitors. Although pSmad8 activity alone could not prevent dKO hfSCs precocious anagen activation, it sustained efficient postnatal differentiation and regeneration of visible hairs. Together, our data suggest a pivotal role for canonical BMP signaling demonstrating distinguished nonoverlapping function of pSmad8 with pSmad1 and pSmad5 in hfSCs regulation and hair morphogenesis but a redundant role in adult hair progenitors differentiation.


[A-mode echography of frontal sinus aplasia].
Twenty-five patients with radiologically proven unilateral frontal sinuses were examined by A-mode ultrasound and the aplastic sinus was compared with the air-filled sinus on the other side. The number of echoes as well as the amplitude and time behaviour of each side were recorded. Statistical evaluation showed no difference in the sonographic parameters between the two sides. Identification of aplastic frontal sinuses by ultrasound is not possible.


Rx with a dose of info.
Information similar to that provided to physicians will be available to patients who get new prescriptions for 10 drugs, as part of a 3-year pilot program of the Food and Drug Administration. The information, in leaflet form, is to be handed out to the customer at the time the prescription is filled. Included will be possible side effect, proper uses for the drug, and any precautions in taking the particular medicine.


Long-term and age-dependent restoration of visual function in a mouse model of CNGB3-associated achromatopsia following gene therapy.
Mutations in the CNGB3 gene account for >50% of all known cases of achromatopsia. Although of early onset, its stationary character and the potential for rapid assessment of restoration of retinal function following therapy renders achromatopsia a very attractive candidate for gene therapy. Here we tested the efficacy of an rAAV2/8 vector containing a human cone arrestin promoter and a human CNGB3 cDNA in CNGB3 deficient mice. Following subretinal delivery of the vector, CNGB3 was detected in both M- and S-cones and resulted in increased levels of CNGA3, increased cone density and survival, improved cone outer segment structure and normal subcellular compartmentalization of cone opsins. Therapy also resulted in long-term improvement of retinal function, with restoration of cone ERG amplitudes of up to 90% of wild-type and a significant improvement in visual acuity. Remarkably, successful restoration of cone function was observed even when treatment was initiated at 6 months of age; however, restoration of normal visual acuity was only possible in younger animals (e.g. 2-4 weeks old). This study represents achievement of the most substantial restoration of visual function reported to date in an animal model of achromatopsia using a human gene construct, which has the potential to be utilized in clinical trials.


[Clinical protocol: audiological assessment of infants in Russian Federation. Part II].
This is the second part of the previously published clinical protocol of audiological assessment in infants. The goal of the protocol is unification approaches to audiological diagnosis of the infants. The following sections were included in the second part of the protocol: behavioral testing in infants, testing sequence, duration of the examination and necessity in follow-up, hearing assessment in special cases (premature children, children with congenital infections, after meningitis, with external ear abnormalities, single-sided deafness, with hydrocephalus and shunts, with auditory neuropathy spectrum disorder, with mild hearing loss and otitis media with effusion), medical report.


Relaxation-compensated difference spin diffusion NMR for detecting 13C-13C long-range correlations in proteins and polysaccharides.
The measurement of long-range distances remains a challenge in solid-state NMR structure determination of biological macromolecules. In 2D and 3D correlation spectra of uniformly (13)C-labeled biomolecules, inter-residue, inter-segmental, and intermolecular (13)C-(13)C cross peaks that provide important long-range distance constraints for three-dimensional structures often overlap with short-range cross peaks that only reflect the covalent structure of the molecule. It is therefore desirable to develop new approaches to obtain spectra containing only long-range cross peaks. Here we show that a relaxation-compensated modification of the commonly used 2D (1)H-driven spin diffusion (PDSD) experiment allows the clean detection of such long-range cross peaks. By adding a z-filter to keep the total z-period of the experiment constant, we compensate for (13)C T1 relaxation. As a result, the difference spectrum between a long- and a scaled short-mixing time spectrum show only long-range correlation signals. We show that one- and two-bond cross peaks equalize within a few tens of milliseconds. Within ~200 ms, the intensity equilibrates within an amino acid residue and a monosaccharide to a value that reflects the number of spins in the local network. With T1 relaxation compensation, at longer mixing times, inter-residue and inter-segmental cross peaks increase in intensity whereas intra-segmental cross-peak intensities remain unchanged relative to each other and can all be subtracted out. Without relaxation compensation, the difference 2D spectra exhibit both negative and positive intensities due to heterogeneous T1 relaxation in most biomolecules, which can cause peak cancellation. We demonstrate this relaxation-compensated difference PDSD approach on amino acids, monosaccharides, a crystalline model peptide, a membrane-bound peptide and a plant cell wall sample. The resulting difference spectra yield clean multi-bond, inter-residue and intermolecular correlation peaks, which are often difficult to resolve in the parent 2D spectra.


Brain SPECT evaluation of patients with pure photosensitive epilepsy.
This study was performed to determine the utility of 99mTc-HMPAO brain SPECT in evaluating patients with pure photosensitive epilepsy. Seven patients (2 boys, 5 girls), aged 8 to 15 yr (mean 11.1 +/- 2.5 yr), were studied. All patients underwent a detailed neurologic examination, interictal and ictal EEGs, CT and/or MRI and SPECT imaging. The baseline SPECT study was performed during the interictal period and the activation study was performed while the patients were having seizures provoked by watching television. The baseline SPECT study showed that six of seven patients had relatively hypoperfused regions in their frontal lobes that could involve the neighboring parietal and temporal regions. The activation study revealed that all seven patients had relative hyperperfusion in these brain regions that were relatively hypoperfused in the baseline study. The side-to-side asymmetry indexes for these visually-interpreted rCBF abnormalities ranged from 3% to 6%. The relatively consistent pattern of frontal rCBF alterations suggests that frontal lobe functions were implicated in the evolution of photosensitivity-related seizures in patients with pure photosensitive epilepsy.


Patterns and perceptions of climate change in a biodiversity conservation hotspot.
Quantifying local people's perceptions to climate change, and their assessments of which changes matter, is fundamental to addressing the dual challenge of land conservation and poverty alleviation in densely populated tropical regions To develop appropriate policies and responses, it will be important not only to anticipate the nature of expected changes, but also how they are perceived, interpreted and adapted to by local residents. The Albertine Rift region in East Africa is one of the world's most threatened biodiversity hotspots due to dense smallholder agriculture, high levels of land and resource pressures, and habitat loss and conversion. Results of three separate household surveys conducted in the vicinity of Kibale National Park during the late 2000s indicate that farmers are concerned with variable precipitation. Many survey respondents reported that conditions are drier and rainfall timing is becoming less predictable. Analysis of daily rainfall data for the climate normal period 1981 to 2010 indicates that total rainfall both within and across seasons has not changed significantly, although the timing and transitions of seasons has been highly variable. Results of rainfall data analysis also indicate significant changes in the intra-seasonal rainfall distribution, including longer dry periods within rainy seasons, which may contribute to the perceived decrease in rainfall and can compromise food security. Our results highlight the need for fine-scale climate information to assist agro-ecological communities in developing effective adaptive management.


Risk of ovarian carcinoma and consumption of vitamins A, C, and E and specific carotenoids: a prospective analysis.
Antioxidant vitamins may decrease risk of cancer by limiting oxidative DNA damage leading to cancer initiation. Few prospective studies have assessed relations between antioxidant vitamins and ovarian carcinoma. The authors prospectively assessed consumption of vitamins A, C, and E and specific carotenoids, as well as fruit and vegetable intake, in relation to ovarian carcinoma risk among 80,326 participants in the Nurses' Health Study who had no history of cancer other than nonmelanoma skin carcinoma. Women reported on known and suspected ovarian carcinoma risk factors including reproductive factors, smoking, and use of vitamin supplements on biennial mailed questionnaires from 1976 to 1996. Food frequency questionnaires were included in 1980, 1984, 1986, and 1990. The authors confirmed 301 incident cases of invasive epithelial ovarian carcinoma during 16 years of dietary follow-up (1980-1996). Pooled logistic regression was used to control for age, oral contraceptive use, body mass index, smoking history, parity, and tubal ligation. The authors observed no association between ovarian carcinoma risk and antioxidant vitamin consumption from foods, or foods and supplements together. The multivariate relative risks (95% confidence intervals [CIs]) for ovarian carcinoma among women in the highest versus lowest quintile of intake were 1.04 (95% CI, 0.72-1.51) for vitamin A from foods and supplements; 1.01 (95% CI, 0.69-1.47) for vitamin C; 0.88 (95% CI, 0.61-1.27) for vitamin E; and 1.10 (95% CI, 0.76-1.59) for beta-carotene. Among users of vitamin supplements, the authors found no evidence of an association between dose or duration of any specific vitamin and ovarian carcinoma risk, although the authors had limited power to assess these relations. No specific fruits or vegetables were associated significantly with ovarian carcinoma risk. The authors found no association between ovarian carcinoma and consumption of total fruits or vegetables, or specific subgroups including cruciferous vegetables, green leafy vegetables, legumes, or citrus fruits. Women who consumed at least 2.5 total servings of fruits and vegetables as adolescents had a 46% reduction in ovarian carcinoma risk (relative risk, 0.54, 95% CI, 0.29-1.03; P value for trend 0.04). These data do not support an important relation between consumption of antioxidant vitamins from foods or supplements, or intake of fruits and vegetables, and incidence of ovarian carcinoma in this cohort. However, modest associations cannot be excluded, and the authors' finding of an inverse association for total fruit and vegetable intake during adolescence raises the possibility that the pertinent exposure period may be much earlier than formerly anticipated.


Phylogenetic relationship of the twenty-one DNA groups of the genus Acinetobacter as revealed by 16S ribosomal DNA sequence analysis.
The inter- and intrageneric relationships of members of the genus Acinetobacter were investigated by performing a comparative sequence analysis of PCR-amplified 16S ribosomal DNAs (rDNAs) from 21 strains representing all of the DNA groups that have been described. Phylogenetic treeing confirmed that Acinetobacter spp. form a coherent cluster within the gamma subdivision of the class Proteobacteria that includes strains with overall levels of 16S rDNA sequence similarity of more than 94%. The analysis of intrageneric relationships suggested that the majority of the strains cluster in five clearly distinguishable clusters, and this conclusion was supported by the results obtained with the different methods used for phylogenetic analysis (i.e., the maximum-likelihood, parsimony, and distance matrix methods). The first cluster contains the representatives of DNA groups 2 (Acinetobacter baumannii) and TU13, whereas the second cluster comprises representatives of DNA groups 3, "Close To TU13," and "between 1 and 3." The representatives of closely related Acinetobacter DNA groups 8 (Acinetobacter twoffii) and 9 belong to the third cluster, which includes the representative of DNA group 6 as well. The fourth cluster is formed by DNA groups BJ15, BJ16, and BJ17, and the fifth cluster comprises DNA groups 1 (Acinetobacter calcoaceticus), BJ14, 10, and 11. Within the fifth cluster the 16S rDNA sequences of DNA group 10 and 11 strains are nearly identical. The representatives of DNA groups 4 (Acinetobacter haemolyticus), 5 (Acinetobacter junii), 7 (Acinetobacter johnsonii), 12 (Acinetobacter radioresistens), TU14, and TU15 form individual branches that are not significantly affiliated with any of the five clusters identified. Apart from the clustering of the most closely related DNA groups, the general topology of the distance dendrogram revealed some discrepancy with previous DNA-DNA hybridization data, which may point to the inadequacy of comparative 16S rDNA sequence analysis for reflecting true evolutionary relationships of closely related bacterial taxa. Important, however, was the presence of unique sequence motifs in each of the 21 different DNA groups studied, which may be useful for rapid differentiation of DNA groups of the genus Acinetobacter.


Hematopoietic rescue via T-cell-dependent, endogenous granulocyte-macrophage colony-stimulating factor induced by the pineal neurohormone melatonin in tumor-bearing mice.
We investigated whether melatonin can affect tumor growth and/or hematopoiesis in mice transplanted with Lewis lung carcinoma and treated with cyclophosphamide or etoposide. These agents were injected i.p. for 5 days at two different cumulative doses (cyclophosphamide, 40 and 160 mg/kg body weight; etoposide, 20 and 40 mg/kg body weight) from day 8 through day 12 after tumor transplantation. Melatonin was injected s.c. at a dose of 1 mg/kg body weight/day, from day 8 throughout the experiments and from days 8 through 12 or from day 13 onwards. Melatonin did not influence tumor growth but selectively counteracted bone marrow toxicity when administered together with the cancer chemotherapy compounds without interfering with their anticancer action. In vitro, melatonin proved to counteract apoptosis in bone marrow cells incubated with etoposide. Such protection was reflected by an increased frequency of granulocyte/macrophage-colony forming units but not of the pluripotent spleen-colony forming units. The effect of melatonin was neutralized by anti-granulocyte/macrophage-colony-stimulating factor monoclonal antibodies. When athymic, T-cell-deficient mice were used as bone marrow donors, melatonin did not exert any protective effect. This suggested that melatonin is able to stimulate the endogenous production of granulocyte/macrophage-colony-stimulating factor via bone marrow T-cells. Due to the well known lack of toxic and undesirable side effects of melatonin, these findings might have a straightforward clinical application.


LRRK2 functions in synaptic vesicle endocytosis through a kinase-dependent mechanism.
Mutations in leucine-rich repeat kinase 2 (LRRK2) are associated with Parkinson's disease, but the precise physiological function of the protein remains ill-defined. Recently, our group proposed a model in which LRRK2 kinase activity is part of an EndoA phosphorylation cycle that facilitates efficient vesicle formation at synapses in the Drosophila melanogaster neuromuscular junctions.Flies harbor only one Lrrk gene, which might encompass the functions of both mammalian LRRK1 and LRRK2. We therefore studied the role of LRRK2 in mammalian synaptic function and provide evidence that knockout or pharmacological inhibition of LRRK2 results in defects in synaptic vesicle endocytosis, altered synaptic morphology and impairments in neurotransmission. In addition, our data indicate that mammalian endophilin A1 (EndoA1,also known as SH3GL2) is phosphorylated by LRRK2 in vitro at T73 and S75, two residues in the BAR domain. Hence, our results indicate that LRRK2 kinase activity has an important role in the regulation of clathrin-mediated endocytosis of synaptic vesicles and subsequent neurotransmission at the synapse.


Characteristics of patients with false- ST-segment elevation myocardial infarction diagnoses.
A subgroup of patients presenting with suspected ST-elevation myocardial infarction (STEMI) have no culprit lesion during coronary angiography (false-positive STEMI). Little is known about patient- and system-related factors that are associated with false-positive STEMI. We evaluated the incidence, correlates, delay, final diagnosis, and outcome of patients with false-positive STEMI. We studied 827 consecutive patients presenting with suspected STEMI between January 2011-September 2012. A false positive STEMI activation was identified in 68 patients (8.2%). Patients with false-positive STEMI were younger (57 vs 63 year; p=0.020), less often had hypercholesterolemia (19 vs 43%; p=0.001), and had a higher heart rate (82 vs 75 bpm; p=0.014). The association between these factors and false-positive STEMI activation persisted in multivariate analysis. The duration of symptoms to call was longer in false-positive STEMI patients (128 vs 83 min; p=0.030), although this did not reach statistical significance in multivariate analysis. Final diagnosis in patients with false-positive STEMI activation was particularly from unknown origin (41%). There were no significant differences in mortality at 30 days and one year between patients with STEMI and false-positive STEMI. The incidence of false-positive STEMI was 8.2% in patients suspected of STEMI. Patients with false-positive STEMI differ from STEMI patients in certain baseline characteristics and in patient delay. Interestingly, absence of coronary disease did not translate into better clinical outcome.


Protein Carriers for Glycoconjugate Vaccines: History, Selection Criteria, Characterization and New Trends.
Currently licensed glycoconjugate vaccines are composed of a carbohydrate moiety covalently linked to a protein carrier. Polysaccharides are T-cell independent antigens able to directly stimulate B cells to produce antibodies. Disease burden caused by polysaccharide-encapsulated bacteria is highest in the first year of life, where plain polysaccharides are not generally immunogenic, limiting their use as vaccines. This limitation has been overcome by covalent coupling carbohydrate antigens to proteins that provide T cell epitopes. In addition to the protein carriers currently used in licensed glycoconjugate vaccines, there is a search for new protein carriers driven by several considerations: (i) concerns that pre-exposure or co-exposure to a given carrier can lead to immune interference and reduction of the anti-carbohydrate immune response; (ii) increasing interest to explore the dual role of proteins as carrier and protective antigen; and (iii) new ways to present carbohydrates antigens to the immune system. Protein carriers can be directly coupled to activated glycans or derivatized to introduce functional groups for subsequent conjugation. Proteins can be genetically modified to pre-determine the site of glycans attachment by insertion of unnatural amino acids bearing specific functional groups, or glycosylation consensus sequences for in vivo expression of the glycoconjugate. A large portion of the new protein carriers under investigation are recombinant ones, but more complex systems such as Outer Membrane Vesicles and other nanoparticles are being investigated. Selection criteria for new protein carriers are based on several aspects including safety, manufacturability, stability, reactivity toward conjugation, and preclinical evidence of immunogenicity of corresponding glycoconjugates. Characterization panels of protein carriers include tests before conjugation, after derivatization when applicable, and after conjugation. Glycoconjugate vaccines based on non-covalent association of carrier systems to carbohydrates are being investigated with promising results in animal models. The ability of these systems to convert T-independent carbohydrate antigens into T-dependent ones, in comparison to traditional glycoconjugates, needs to be assessed in humans.


Improvement of the clinical skills of nurse anesthesia students using mini-clinical evaluation exercises in Iran: a randomized controlled study.
The present study aimed to investigate the effect of a mini-clinical evaluation exercise (CEX) assessment on improving the clinical skills of nurse anesthesia students at Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran. This study started on November 1, 2022, and ended on December 1, 2022. It was conducted among 50 nurse anesthesia students divided into intervention and control groups. The intervention group's clinical skills were evaluated 4 times using the mini-CEX method. In contrast, the same skills were evaluated in the control group based on the conventional method—that is, general supervision by the instructor during the internship and a summative evaluation based on a checklist at the end of the course. The intervention group students also filled out a questionnaire to measure their satisfaction with the miniCEX method. The mean score of the students in both the control and intervention groups increased significantly on the post-test (P<0.0001), but the improvement in the scores of the intervention group was significantly greater compared with the control group (P<0.0001). The overall mean score for satisfaction in the intervention group was 76.3 out of a maximum of 95. The findings of this study showed that using mini-CEX as a formative evaluation method to evaluate clinical skills had a significant effect on the improvement of nurse anesthesia students' clinical skills, and they had a very favorable opinion about this evaluation method.


Multihormonal induction of alpha 2u-globulin in an established rat hepatoma cell line.
A subclone of the FU5-5 rat hepatoma cell line has been isolated which is inducible more than several hundred fold for the 20,000 dalton form of the major rat urinary protein alpha 2u-globulin. The basal relative synthetic rate (RSR) in growth medium containing 10% fetal calf serum was less than 2 X 10(-6) of total protein synthesis. Both dexamethasone and insulin were necessary for induction, and yielded a maximum induced RSR of 4-8 X 10(-3). Triiodothyronine (T3), dihydrotestosterone (DHT), rat growth hormone (GH), and estrogen, all of which have been shown to influence the induction of alpha 2u-globulin in the intact rat, were without effect on the cell line. A factor present in fetal calf serum was also necessary for maximum induction, since dexamethasone plus insulin in serum-free medium raised the RSR to only 3 X 10(-5); exogenous T3, GH, and DHT could not substitute for this serum factor. The kinetics of induction by dexamethasone were slow, with a lag of approximately 48 hr followed by a period of increasing RSR for 6-20 days. Removal of dexamethasone from induced cells led to an exponential decline in the RSR (t 1/2 15 hr). The concentrations of dexamethasone and insulin that could yield half maximum induction were 5 X 10(-8)M and 3 X 10(-11)M, respectively. Higher concentrations of insulin, although still in physiological range (10(-9)M), inhibited induction. At yet higher insulin levels, beyond the physiological range, alpha 2u-globulin synthesis returned to maximum values. The lack of DHT, T3, and GH requirement for alpha 2u-globulin induction in this cell line may mean that a regulatory aberrancy has occurred in this transformed cell line, or, alternatively, that these hormones act indirectly in the intact animal. This cell line should prove useful for the study of the molecular events associated with alpha 2u-globulin induction and for genetic approaches to the problem of multihormonal regulation of gene expression.


The Report of the 2016-2017 Advocacy Standing Committee.
Based on the growing importance of community engagement and the recognition of its importance by the American Association of Colleges of Pharmacy (AACP), the committee offers several examples of community engagement activities for consideration and replication by our academy and beyond. These activities, including those of winning institutions of the Lawrence J. Weaver Transformational Community Engagement Award, can be mapped to the core components of community engagement presented in Table 1. The committee, using an implementation readiness framework, provides the reader with insight into the challenges that may impact successful community engagement and encourages our academy to continue its work to support faculty capacity in this area. Toward that end, the committee offers a policy statement that encourages schools and colleges of pharmacy to have an office or designate a faculty member whose focus is specifically on community engagement. The committee also offers a recommendation that the core components be included in the criteria for the Weaver Award.


Sex, Body Image, and Relationships: A BRIGHTLIGHT Workshop on Information and Support Needs of Adolescents and Young Adults.
Discovering sexuality and romantic relationships are important development milestones in adolescence and young adulthood. A cancer diagnosis imposes obstacles for young people such as changes in their sexual function due to the disease and/or side effects of treatment, body image concerns, and interpersonal relationship difficulties. This can cause psychological distress and can impact on quality of life. We aimed to explore sexual health information and support needs of adolescents and young adults with cancer. Five young people aged 16-24 years, with a previous cancer diagnosis when aged 13-22 years, attended an in-depth 4-hour workshop. The framework approach was used to analyze workshop transcripts. Three overarching themes emerged: (i) information sharing; (ii) contexts and relationships (influencing factors); and (iii) information sharing preferences. Information shared by healthcare professionals was focused on a medicalized view of sex with symptoms, infection control, and protected sex at its core. Young people had unanswered questions related to sexual function, the impact of cancer and how to manage it, and about pleasure, body image, and relationships. Parents' presence at clinical consultations inhibited discussions about sex. Young people wanted professionals who were comfortable to talk about sex with them. Young people exhibited significant unmet needs around information provision on sex, body image, and relationships. They wanted information to be given by professionals and access to online resources. Development of training for professionals and resources to support young people requires further work.


The Use of Ultrasound Imaging in the External Beam Radiotherapy Workflow of Prostate Cancer Patients.
External beam radiotherapy (EBRT) is one of the curative treatment options for prostate cancer patients. The aim of this treatment option is to irradiate tumor tissue, while sparing normal tissue as much as possible. Frequent imaging during the course of the treatment (image guided radiotherapy) allows for determination of the location and shape of the prostate (target) and of the organs at risk. This information is used to increase accuracy in radiation dose delivery resulting in better tumor control and lower toxicity. Ultrasound imaging is harmless for the patient, it is cost-effective, and it allows for real-time volumetric organ tracking. For these reasons, it is an ideal technique for image guidance during EBRT workflows. Review papers have been published in which the use of ultrasound imaging in EBRT workflows for different cancer sites (prostate, breast, etc.) was extensively covered. This new review paper aims at providing the readers with an update on the current status for prostate cancer ultrasound guided EBRT treatments.


Serum indoxyl sulfate is associated with vascular disease and mortality in chronic kidney disease patients.
As a major component of uremic syndrome, cardiovascular disease is largely responsible for the high mortality observed in chronic kidney disease (CKD). Preclinical studies have evidenced an association between serum levels of indoxyl sulfate (IS, a protein-bound uremic toxin) and vascular alterations. The aim of this study is to investigate the association between serum IS, vascular calcification, vascular stiffness, and mortality in a cohort of CKD patients. One-hundred and thirty-nine patients (mean +/- SD age: 67 +/- 12; 60% male) at different stages of CKD (8% at stage 2, 26.5% at stage 3, 26.5% at stage 4, 7% at stage 5, and 32% at stage 5D) were enrolled. Baseline IS levels presented an inverse relationship with renal function and a direct relationship with aortic calcification and pulse wave velocity. During the follow-up period (605 +/- 217 d), 25 patients died, mostly because of cardiovascular events (n = 18). In crude survival analyses, the highest IS tertile was a powerful predictor of overall and cardiovascular mortality (P = 0.001 and 0.012, respectively). The predictive power of IS for death was maintained after adjustment for age, gender, diabetes, albumin, hemoglobin, phosphate, and aortic calcification. The study presented here indicates that IS may have a significant role in the vascular disease and higher mortality observed in CKD patients.


The Cardiorespiratory Demands of Treadmill Walking with and without the Use of Ekso GT™ within Able-Bodied Participants: A Feasibility Study.
Individuals with neurological impairments tend to lead a predominantly sedentary lifestyle due to impaired gait function and mobility. This may be detrimental to health by negatively impacting cardiorespiratory fitness and muscular strength, and increasing the risk of developing secondary health problems. Powered exoskeletons are assistive devices that may aid neurologically impaired individuals in achieving the World Health Organisation's (WHO) physical activity (PA) guidelines for health. Increased PA should elicit a sufficient cardiorespiratory stimulus to provide health benefits to exoskeleton users. This study examined the cardiorespiratory demands of treadmill walking with and without the Ekso GT™ among able-bodied participants. The Ekso GT™ is a powered exoskeleton that enables individuals with neurological impairments to walk by supporting full body mass with motors attached at the hip and knee joints to generate steps. This feasibility study consisted of one group of healthy able-bodied individuals (n = 8). Participants completed two 12 min treadmill walking assessments, one with and one without the Ekso GT™ at the same fixed speed. Throughout each walking bout, various cardiorespiratory parameters, namely, volume of oxygen per kilogram (kg) of body mass (V˙O2·kg-1), volume of carbon dioxide per kg of body mass (V˙CO2·kg-1), respiratory exchange ratio (RER), ventilation (V˙E), heart rate (HR), and rate of perceived exertion (RPE), were recorded. Treadmill walking with Ekso GT™ elevated all recorded measurements to a significantly greater level (p ≤ 0.05) (except RER at 1 km·h-1; p = 0.230) than treadmill walking without the Ekso GT™ did at the same fixed speed. An increased cardiorespiratory response was recorded during treadmill walking with the exoskeleton. Exoskeleton walking may, therefore, be an effective method to increase PA levels and provide sufficient stimulus in accordance with the PA guidelines to promote cardiorespiratory fitness and subsequently enhance overall health.


Fluorescence in situ hybridization analysis of 12;21 translocation in Japanese childhood acute lymphoblastic leukemia.
Fluorescence in situ hybridization (FISH) analysis was applied to detect t(12;21) using two yeast artificial chromosome probes and cosmid probes covering the TEL(ETV6) and the AML1 gene to clarify the incidence of abnormality of t(12;21) in Japanese childhood acute lymphoblastic leukemia (ALL). We detected seven TEL/AML1 fusion positive patients (9.5%), all of whom were diagnosed as B-lineage ALL, among 74 childhood ALL. On the other hand, no TEL/AML1 fusion positive patients were found among 37 adult ALL. The incidence among Japanese seemed to be lower than that among other nations. Of the seven patients with the TEL/AML1 fusion, five exhibited normal karyotype, one was t(8;12)(q11;p13), i(21q) and the remaining one exhibited a near-triploid karyotype in conventional G-banding. The FISH method clearly demonstrated that all patients with the TEL/AML1 fusion had subpopulations of leukemic cells with deletion of the normal TEL allele, which is significant for understanding the progression of leukemia with t(12;21).


Comparison of computed tomographic and pathologic findings in 17 dogs with primary adrenal neoplasia.
The CT appearance of canine adrenal masses has been reported, but associations between imaging features and pathologic features of these lesions have not been investigated in detail. The purpose of this study was to test associations between different types of adrenal neoplasia and their CT and pathologic features. A retrospective cross-sectional study was performed and inclusion criteria were histologic diagnosis of primary adrenal neoplasia, contrast-enhanced CT examination of the abdomen and surgical resection of the mass or necropsy examination. For all included dogs, CT images and histopathologic specimens were reviewed independently by two veterinary radiologists and a veterinary pathologist, respectively. Seventeen dogs met inclusion criteria. Diagnoses were adenocarcinoma in nine (53%) dogs, pheochromocytoma in five (29%) dogs, and adenoma in three (18%) dogs. Pheochromocytoma was associated with CT signs of vascular invasion (likelihood ratio = 4.8, 95% CI = 1.3-18.3, P = 0.03) and macroscopic vascular invasion (likelihood ratio = 9.6, 95% CI = 1.4-65.9, P = 0.02). There was excellent agreement between signs of vascular invasion in CT images and vascular invasion at surgery or necropsy (kappa = 0.86, P = 0.001). A peripheral contrast-enhancing rim in delayed postcontrast CT images was associated with fibrous encapsulation of the tumor (kappa = 0.53, P = 0.05), and a heterogeneous pattern of contrast distribution in delayed postcontrast CT images was associated with adrenal hemorrhage or infarction on histological examination (kappa = 0.45, P = 0.05). Findings indicated that CT enabled assessment of adrenal neoplasia features that reflected their biological behavior and pathological findings, however overlapping characteristics between tumor types limited the potential for reliably distinguishing them based on CT alone.


Infections, bacterial resistance, and antimicrobial stewardship: the emerging role of hospitalists.
The care of patients with serious infections both within and outside healthcare settings is increasingly complicated by the high prevalence of resistant or multidrug-resistant (MDR) pathogens. Moreover, infections caused by MDR versus susceptible bacteria or other pathogens are associated with significantly higher mortality, length of hospital stay, and healthcare costs. Antimicrobial misuse or overuse is the primary driver for development of antimicrobial resistance, suggesting that better use of antimicrobials will translate into improved patient outcomes, more efficient use of hospital resources, and lowered healthcare costs. Antimicrobial stewardship refers to the various practices and procedures utilized to optimize antimicrobial use. The primary goal of antimicrobial stewardship is to improve patient outcomes and lower antimicrobial resistance and other unintended consequences of antimicrobial therapy. Secondary goals are to reduce length of hospital stays and healthcare-related costs. Hospitalists are increasingly involved in the care of hospitalized patients throughout the United States. Expertise in managing conditions requiring hospitalization, and experience in quality improvement across a wide range of clinical conditions, make hospitalists well positioned to participate in the development and implementation of hospital-based antimicrobial stewardship programs designed to improve patient outcomes, reduce antimicrobial resistance, and provide more efficient and lower-cost hospital care. Journal of Hospital Medicine 2012;7:S34-S43. © 2012 Society of Hospital Medicine.


Quantum and isotope effects in lithium metal.
The crystal structure of elements at zero pressure and temperature is the most fundamental information in condensed matter physics. For decades it has been believed that lithium, the simplest metallic element, has a complicated ground-state crystal structure. Using synchrotron x-ray diffraction in diamond anvil cells and multiscale simulations with density functional theory and molecular dynamics, we show that the previously accepted martensitic ground state is metastable. The actual ground state is face-centered cubic (fcc). We find that isotopes of lithium, under similar thermal paths, exhibit a considerable difference in martensitic transition temperature. Lithium exhibits nuclear quantum mechanical effects, serving as a metallic intermediate between helium, with its quantum effect-dominated structures, and the higher-mass elements. By disentangling the quantum kinetic complexities, we prove that fcc lithium is the ground state, and we synthesize it by decompression.


Selective spatiotemporal induction of matrix metalloproteinase-2 and matrix metalloproteinase-9 transcription after myocardial infarction.
Myocardial remodeling after myocardial infarction (MI) is associated with increased levels of the matrix metalloproteinases (MMPs). Levels of two MMP species, MMP-2 and MMP-9, are increased after MI, and transgenic deletion of these MMPs attenuates post-MI left ventricular (LV) remodeling. This study characterized the spatiotemporal patterns of gene promoter induction for MMP-2 and MMP-9 after MI. MI was induced in transgenic mice in which the MMP-2 or MMP-9 promoter sequence was fused to the beta-galactosidase reporter, and reporter level was assayed up to 28 days after MI. Myocardial localization with respect to cellular sources of MMP-2 and MMP-9 promoter induction was examined. After MI, LV diameter increased by 70% (P < 0.05), consistent with LV remodeling. beta-Galactosidase staining in MMP-2 reporter mice was increased by 1 day after MI and increased further to 64 +/- 6% of LV epicardial area by 7 days after MI (P < 0.05). MMP-2 promoter activation occurred in fibroblasts and myofibroblasts in the MI region. In MMP-9 reporter mice, promoter induction was detected after 3 days and peaked at 7 days after MI (53 +/- 6%, P < 0.05) and was colocalized with inflammatory cells at the peri-infarct region. Although MMP-2 promoter activation was similarly distributed in the MI and border regions, activation of the MMP-9 promoter was highest at the border between the MI and remote regions. These unique findings visually demonstrated that activation of the MMP-2 and MMP-9 gene promoters occurs in a distinct spatial relation with reference to the MI region and changes in a characteristic time-dependent manner after MI.


Disrupted Peyer's Patch Microanatomy in COVID-19 Including Germinal Centre Atrophy Independent of Local Virus.
Confirmed SARS-coronavirus-2 infection with gastrointestinal symptoms and changes in microbiota associated with coronavirus disease 2019 (COVID-19) severity have been previously reported, but the disease impact on the architecture and cellularity of ileal Peyer's patches (PP) remains unknown. Here we analysed post-mortem tissues from throughout the gastrointestinal (GI) tract of patients who died with COVID-19. When virus was detected by PCR in the GI tract, immunohistochemistry identified virus in epithelium and lamina propria macrophages, but not in lymphoid tissues. Immunohistochemistry and imaging mass cytometry (IMC) analysis of ileal PP revealed depletion of germinal centres (GC), disruption of B cell/T cell zonation and decreased potential B and T cell interaction and lower nuclear density in COVID-19 patients. This occurred independent of the local viral levels. The changes in PP demonstrate that the ability to mount an intestinal immune response is compromised in severe COVID-19, which could contribute to observed dysbiosis.


Prevalence of cognitive impairment in peritoneal dialysis patients and associated factors.
Cognitive impairment (CI) in patients with chronic kidney disease, including those treated with renal replacement therapy (RRT) is a growing problem worldwide. The study aimed to assess the prevalence of CI and associated factors in patients undergoing peritoneal dialysis (PD). In this cross-sectional study, 18 consecutive patients with PD therapy and 15 controls were evaluated for CI using the Addenbrooke's Cognitive Examination III (ACE III) test. The prevalence of CI was 33% in patients and 27% in the control group and was not statistically significant. A higher prevalence of CI was found in subjects aged ≥65 years old than in those <65years old (p = 0.02), but only in the control group. The prevalence of CI in PD patients over and under 65 years of age did not differ statistically significantly (p = 0.12). Memory and verbal fluency were the most affected cognitive domain in PD patients with CI (p = 0.00, p = 0.04, respectively). There was a significant correlation between higher-educated PD patients and the ACE III test results. The duration of dialysis did not affect the results of the cognitive screening test. Cognitive impairment is a growing problem in the course of chronic kidney disease and dialysis therapy. It seems that cognitive problems may occur in patients undergoing peritoneal dialysis at a younger age than in the general population with particularly affected memory and verbal fluency. Higher-educated patients score better on the cognitive screening test.


Relationship between SYNTAX score and myocardial viability in ischemic cardiomyopathy.
The SYNTAX score (SS) is not just a measure of the severity of coronary artery disease, but also complexity. The aim of this study was to evaluate the relationship between the SS and myocardial viability/non-viability assessed by positron emission tomography (PET) in patients with ischemic cardiomyopathy (IC). A total of 107 IC patients who had undergone PET were enrolled in the study. The patients were divided into two groups according to the presence or absence of viable myocardium. SS was analyzed from recorded conventional coronary angiographies. Patients with a non-viable myocardium (n=21; 19.6%) had a significantly higher SS compared to those with a viable myocardium (17.6±3.7 vs. 14.1±5.2, respectively; p=0.004). Point-biserial correlation coefficient analysis indicated that the presence of myocardial non-viability was mildly correlated with a higher SS (rpb=-0.28, p=0.004). In multivariate logistic regression analysis, the SS was identified as the sole independent predictor of myocardial non-viability (odds ratio [OR]: 1.164, 95% confidence interval [CI]: 1.044-1.297; p =0.006]. Receiver operating characteristic analysis revealed a cutoff point of 16 for predicting a non-viable myocardium (area under curve: 0.71, 95% CI: 0.61-0.82) with a sensitivity of 76.2% and a specificity of 61.6%. The results of the present study indicates that a high SS is associated with the presence of a non-viable myocardium in IC patients.


Perceptions, motivations, and concerns about living organ donation among people living with HIV.
Recent changes to United States law now permit people living with HIV (PLWH) to donate organs to HIV-infected (HIV+) recipients under research protocols. PLWH may have unique motivations for and concerns about living donation and understanding them is critical to ensuring the integrity of this novel approach to organ transplantation. We conducted in-depth interviews with PLWH from an urban HIV clinic who had previously indicated their willingness to be a living donor. Interviews elicited information on their motivations, perceived benefits, and concerns regarding living donation. Codes were identified inductively and then organized into themes and subthemes. Two coders independently analyzed the interviews and reconciled differences in coding by consensus. Thematic saturation was reached after 20 interviews. Motivations for living donation among PLWH included an altruistic desire to help others as well as HIV-specific motivations including solidarity with potential recipients and a desire to overcome HIV-related stigma. Perceived benefits of living donation included gratification from saving or improving the recipient's life and conferring a sense of normalcy for the HIV+ donor. Concerns about donation included the possibility of a prolonged recovery period, organ failure, and transmission of another strain of the virus to the recipients. PLWH had unique motivations, perceived benefits, and concerns about living donation in addition to those previously identified in the general population. These unique factors should be addressed in research protocols, informed consent processes, and the education and training of independent living donor advocates so that these endeavors are ethically sound.


Three-Dimensional Network of Cation-Cation-Bound Neptunyl(V) Squares: Synthesis and in Situ Raman Spectroscopy Studies.
Cation-cation interactions (CCIs) are an essential feature of actinyl chemistry, particularly neptunyl(V). To better understand the formation mechanisms of CCIs, the crystallization process of Np(V) CCI compounds has been explored during the evaporation of acidic Np(V) stock solutions using X-ray diffraction and both ex situ and in situ Raman spectroscopy. At least four Np solid products have been isolated from evaporation of the same Np(V) acidic solution. In situ evaporation using a continuous wave laser (532 nm) as a local heat source produced similar solid products to ex situ experiments with matching Raman signatures. The formation of these products is highly dependent on the evaporation conditions. Slower evaporation appears to favor the formation of a new neptunyl(V) compound, (NpO2)Cl(H2O)2 (1), over other solid products. The structure of 1 features a three-dimensional network of NpO2(+) cations, where neighboring Np(V) ions are only connected to each other through CCIs in a square arrangement. The O═Np═O stretching region shows similar Raman bands in both the solids and solution suggesting that CCIs between Np(V) cations exist prior to crystallization. These results provide new insight into the formation mechanism of Np(V) CCI compounds from solutions.