Tumor-specific cytotoxic T cell clones from patients with breast and pancreatic adenocarcinoma recognize EBV-immortalized B cells transfected with polymorphic epithelial mucin complementary DNA.
Human breast and pancreatic adenocarcinomas are tumors of ductal epithelial cell origin and as such produce and express on their surface polymorphic epithelial cell mucin encoded by the MUC 1 gene. We have previously reported that tumor-specific cytotoxic T cells derived from patients bearing such tumors recognize specific epitopes on the mucin polypeptide core. This recognition was not MHC-restricted because of the highly repetitive sequence of the polypeptide core, which allows simultaneous recognition of many identical epitopes, and cross-linking and aggregation of TCR on mucin-specific T cells. Those studies were performed with limited numbers of tumor cells or allogeneic tumor cell lines. A renewable source of autologous cells presenting this Ag was necessary to further explore mucin-specific immunity. We report here successful establishment and functional analysis of mucin-specific CTL lines and clones derived from breast and pancreatic cancer patients, using either autologous or allogeneic mucin-transfected B cells as Ag. Our results demonstrate that transfection of autologous or allogeneic B cells with mucin confers upon them tumor Ag-presenting ability as well as susceptibility to lysis by mucin-specific CTL. Transfection of APC with this or any other human tumor Ag that may be molecularly defined in the future provides a unique and powerful tool with which to examine the ability of a tumor-associated Ag to stimulate T cell responses.


Temporal eye movement strategies during naturalistic viewing.
The deployment of eye movements to complex spatiotemporal stimuli likely involves a variety of cognitive factors. However, eye movements to movies are surprisingly reliable both within and across observers. We exploited and manipulated that reliability to characterize observers' temporal viewing strategies while they viewed naturalistic movies. Introducing cuts and scrambling the temporal order of the resulting clips systematically changed eye movement reliability. We developed a computational model that exhibited this behavior and provided an excellent fit to the measured eye movement reliability. The model assumed that observers searched for, found, and tracked a point of interest and that this process reset when there was a cut. The model did not require that eye movements depend on temporal context in any other way, and it managed to describe eye movements consistently across different observers and two movie sequences. Thus, we found no evidence for the integration of information over long time scales (greater than a second). The results are consistent with the idea that observers employ a simple tracking strategy even while viewing complex, engaging naturalistic stimuli.


Characterization of a novel Taenia solium oncosphere antigen.
Infections due to Taenia solium in humans (taeniasis/cysticercosis) remain a complex health problem, particularly in developing countries. We identified two oncosphere proteins that might protect the porcine intermediate host against cysticercosis and therefore help prevent disease in humans. One of these proteins was further identified by two-dimensional gel electrophoresis and micro-sequencing. The gene encoding this protective protein was also identified, cloned and characterized. The native 31.5 kDa protein Tso31 has four variants at the cDNA level. The longest sequence from which the others seem to derive, encodes a 253 amino acid peptide. The predicted protein has a molecular weight of 25.1 kDa, one putative N-glycosylation site, two fibronectin type III domains, and one C terminal transmembrane domain. The gene structure of the protein consists of four exons and three introns. The finding of one gene and four different cDNAs for Tso31 suggests the existence of a possible mechanism of differential splicing in this parasite. The Tso31 protein is exclusive to T. solium oncospheres with a putative protein structure of an extra-cellular receptor-like protein. The Tso31 protein was expressed as a recombinant protein fused to GST and tested in a vaccine to determine its effectiveness in protecting pigs against cysticercosis. Only two pigs out of eight vaccinated were protected and although the total median number of cyst decreased in vaccinated pigs compared to controls this decrease was not statistically significant (P = 0.09).


Novel Approaches in Primary Cardiovascular Disease Prevention: The HOPE-3 Trial Rationale, Design, and Participants' Baseline Characteristics.
Cholesterol and blood pressure (BP) can be effectively and safely lowered with statin drugs and BP-lowering drugs, reducing major cardiovascular (CV) events by 20%-30% within 5 years in high-risk individuals. However, there are limited data in lower-risk populations. The Heart Outcomes Prevention Evaluation-3 (HOPE-3) trial is evaluating whether cholesterol lowering with a statin drug, BP lowering with low doses of 2 antihypertensive agents, and their combination safely reduce major CV events in individuals at intermediate risk who have had no previous vascular events and have average cholesterol and BP levels. A total of 12,705 women 65 years or older and men 55 years or older with at least 1 CV risk factor, no known CV disease, and without any clear indication or contraindication to the study drugs were randomized to rosuvastatin 10 mg/d or placebo and to candesartan/hydrochlorothiazide 16/12.5 mg/d or placebo (2 × 2 factorial design) and will be followed for a mean of 5.8 years. The coprimary study outcomes are the composite of CV death, nonfatal myocardial infarction (MI), and nonfatal stroke and the composite of CV death, nonfatal MI, nonfatal stroke, resuscitated cardiac arrest, heart failure, and arterial revascularization. Participants were recruited from 21 countries in North America, South America, Europe, Asia, and Australia. Mean age at randomization was 66 years and 46% were women. The HOPE-3 trial will provide new information on cholesterol and BP lowering in intermediate-risk populations with average cholesterol and BP levels and is expected to inform approaches to primary prevention worldwide (HOPE-3 ClinicalTrials.gov NCT00468923).


Wild orangutan males plan and communicate their travel direction one day in advance.
The ability to plan for the future beyond immediate needs would be adaptive to many animal species, but is widely thought to be uniquely human. Although studies in captivity have shown that great apes are capable of planning for future needs, it is unknown whether and how they use this ability in the wild. Flanged male Sumatran orangutans (Pongo abelii) emit long calls, which females use to maintain earshot associations with them. We tested whether long calls serve to communicate a male's ever-changing predominant travel direction to facilitate maintaining these associations. We found that the direction in which a flanged male emits his long calls predicts his subsequent travel direction for many hours, and that a new call indicates a change in his main travel direction. Long calls given at or near the night nest indicate travel direction better than random until late afternoon on the next day. These results show that male orangutans make their travel plans well in advance and announce them to conspecifics. We suggest that such a planning ability is likely to be adaptive for great apes, as well as in other taxa.


Changes in parvalbumin immunoreactive retinal ganglion cells and amacrine cells after optic nerve injury.
Parvalbumin (PARV) is a Ca(2+)-binding protein that may offer resistance to cell death as it primarily functions to maintain Ca(2+) homeostasis. The purpose of this study was to investigate whether PARV expressing retinal ganglion cells (RGCs) would be more resistant to cell death than RGCs that do not express PARV. RGCs in Sprague-Dawley rats were retrogradely labeled with Fluorogold (FG). After 2-28 days following an optic nerve crush (ONC) injury immunohistochemistry was performed on the sections using antibodies against PARV and markers of RGCs. The proportion of retinal ganglion cell layer cells labeled with PARV colocalized with FG or Brn3a and labeled only with PARV (displaced amacrine cells; dACs) were analyzed. PARV staining intensity was measured in ACs, dACs, and RGCs. Double labeling studies revealed that 49% of RGCs and 22% of dACs expressed PARV. There was an immediate reduction in RGC PARV staining after ONC but the overall rate of cell death after 28 days was similar in PARV and non-PARV expressing RGCs. There was no change in PARV AC or dAC number or staining intensity. Although this study suggests that there is no selective survival of the subpopulation of RGCs that contain PARV, there is down-regulation of PARV expression by these RGCs. This suggests that down-regulation of PARV may contribute to RGC death due to a compromised Ca(2+) buffering capacity. Maintaining PARV expression after injury could be an important neuroprotective strategy to improve RGC survival after injury.


Hepatic adenomas with synchronous or metachronous fibrolamellar carcinomas: both are characterized by LFABP loss.
Rare hepatic adenomas are associated with synchronous or metachronous fibrolamellar carcinomas. The morphology of these adenomas has not been well described and they have not been subclassifed using the current molecular classification schema. We examined four hepatic adenomas co-occurring with or preceding a diagnosis of fibrolamellar carcinoma in three patients. On histological examination, three of the adenomas showed the typical morphology of HNF1-α inactivated adenomas, whereas one showed a myxoid adenoma morphology. All of the adenomas were negative for PRKACA rearrangements by Fluorescence in situ Hybridization (FISH) analysis. All four of the adenomas showed complete loss or significant reduction of liver fatty acid binding protein (LFABP) expression by immunohistochemistry. Interestingly, the fibrolamellar carcinomas in each case also showed loss of LFABP by immunohistochemistry. One of the fibrolamellar carcinomas was negative for PRKACA rearrangements by FISH, whereas the others were positive. To investigate if LFBAP loss is typical of fibrolamellar carcinomas in general, an additional cohort of tumors was studied (n=19). All 19 fibrolamellar carcinomas showed the expected PRKACA rearrangements and immunostains showed loss of LFABP in each case, consistent with HNF1-α inactivation. To validate this observation, mass spectrometry-based proteomics was performed on tumor-normal pairs of six fibrolamellar carcinomas and showed an average 10-fold reduction in LFABP protein levels, compared with matched normal liver tissue. In conclusion, hepatic adenomas co-occurring with fibrolamellar carcinomas show LFABP loss and are negative for PRKACA rearrangements, indicating they are genetically distinct lesions. These data also demonstrate that LFABP loss, which characterizes HNF1-α inactivation, is a consistent feature of fibrolamellar carcinoma, indicating HNF1-α inactivation is an important event in fibrolamellar carcinoma pathogenesis.


The odyssey of cobaloximes for catalytic H2 production and their recent revival with enzyme-inspired design.
Cobaloxime complexes gained attention for their intrinsic ability of catalytic H2 production despite their initial emergence as a vitamin B12 model. The simple, robust, and synthetically manoeuvrable cobaloxime core represents a model catalyst molecule for the investigation of optimal conditions for both photo- and electrocatalytic H2 production catalytic assemblies. Cobaloxime is one of the rare catalysts that finds equal applications in the analysis of homogeneous and heterogeneous catalytic conditions. However, the poor aqueous solubility and long-term instability of cobaloximes have severely impeded their growth. Lately, interest in the cobaloxime-based catalysts has been resuscitated with the rational use of extended enzymatic features. This unique enzyme-inspired catalyst design strategy has instigated the formation of a new genre of cobaloxime molecules that exhibit enhanced photo- and electrocatalytic H2 evolution with improved aqueous and air stability.


Automated tissue segmentation and blind recovery of (1)H MRS imaging spectral patterns of normal and diseased human brain.
Constrained non-negative matrix factorization (cNMF) with iterative data selection is described and demonstrated as a data analysis method for fast and automatic recovery of biochemically meaningful and diagnostically specific spectral patterns of the human brain from (1)H MRS imaging ((1)H MRSI) data. To achieve this goal, cNMF decomposes in vivo multidimensional (1)H MRSI data into two non-negative matrices representing (a) the underlying tissue-specific spectral patterns and (b) the spatial distribution of the corresponding metabolite concentrations. Central to the proposed approach is automatic iterative data selection which uses prior knowledge about the spatial distribution of the spectra to remove voxels that are due to artifacts and undesired metabolites/tissues such as the strong lipid and water components. The automatic recovery of diagnostic spectral patterns is demonstrated for long-TE (1)H MRSI data on normal human brain, multiple sclerosis, and serial brain tumor. The results show the ability of cNMF with iterative data selection to automatically and simultaneously recover tissue-specific spectral patterns and achieve segmentation of normal and diseased human brain tissue, concomitant with simplification of information content. These features of cNMF, which permit rapid recovery, reduction and interpretation of the complex diagnostic information content of large multi-dimensional spectroscopic imaging data sets, have the potential to enhance the clinical utility of in vivo(1)H MRSI.


The Effects of Discrimination Are Associated With Cigarette Smoking Among Black Males.
Previous research has demonstrated that experiencing interpersonal discrimination is associated with cigarette smoking. Few studies have examined the relationship between the effects of physical and emotional discrimination and cigarette usage, and none have examined this relationship among Black men. The aim of this study was to examine the association between the effects of physical and emotional discrimination and cigarette smoking. Data from the Indiana Black Men's Health Study, a community-based sample of adult Black men, was used to conduct multivariate logistic regression to examine the relationship between the physical and emotional effects of discrimination and smoking, net of healthcare and workplace discrimination, age, education, household income, and being married. After adjusting for having an emotional response to discrimination, health care and workplace discrimination, age, education, household income, and being married, males who had a physical response to discrimination (e.g., upset stomach or headache) had higher odds of cigarette use (odds ratio (OR): 1.95, 95% confidence interval (CI): 1.15-3.30) than men who did not have a physical response to discrimination. Findings from the study suggest that Black males may use cigarette smoking as a means to mitigate the stress associated with experiences of discrimination. Future research is needed further to explore if and how Black males use cigarette smoking to cope with unfair treatment.


Astrocytes and Microglia-Mediated Immune Response in Maladaptive Plasticity is Differently Modulated by NGF in the Ventral Horn of the Spinal Cord Following Peripheral Nerve Injury.
Reactive astrocytes and activated microglia are the key players in several pathophysiologic modifications of the central nervous system. We used the spared nerve injury (SNI) of the sciatic nerve to induce glial maladaptive response in the ventral horn of lumbar spinal cord and examine its role in the remodeling of the tripartite synapse plasticity. Imaging the ventral horn revealed that SNI was associated with both an early microglial and astrocytic activation, assessed, respectively, by analysis of Iba1 and GFAP expression. Microglia, in particular, localized peculiarly surrounding the motor neurons somata. Perineuronal astrocytes, which play a key role in maintaining the homeostasis of neuronal circuitry, underwent a substantial phenotypic change following peripheral axotomy, producing reactive gliosis. The gliosis was associated with the reduction of glial aminoacid transporters (GLT1 and GlyT1) and increase of neuronal glutamate transporter EAAC1. Although the expression of GABAergic neuronal marker GAD65/67 showed no change, glutamate increase, as demonstrated by HPLC analysis, shifted the excitatory/inhibitory balance as showed by the net increase of the glutamate/GABA ratio. Moreover, endogenous NGF levels were altered in SNI animals and not restored by the intrathecal NGF administration. This treatment reverted phenotypic changes associated with reactive astrocytosis, but failed to modify microglia activation. These findings on one hand confirm the correlation between gliopathy and maladaptive plasticity of the spinal synaptic circuitry, on the other hand add new data concerning the complex peculiar behavior of different glial cells in neuronal degenerative processes, defining a special role of microglia in sustaining the inflammatory response.


Sequential sensory and decision processing in posterior parietal cortex.
Decisions about the behavioral significance of sensory stimuli often require comparing sensory inference of what we are looking at to internal models of what we are looking for. Here, we test how neuronal selectivity for visual features is transformed into decision-related signals in posterior parietal cortex (area LIP). Monkeys performed a visual matching task that required them to detect target stimuli composed of conjunctions of color and motion-direction. Neuronal recordings from area LIP revealed two main findings. First, the sequential processing of visual features and the selection of target-stimuli suggest that LIP is involved in transforming sensory information into decision-related signals. Second, the patterns of color and motion selectivity and their impact on decision-related encoding suggest that LIP plays a role in detecting target stimuli by comparing bottom-up sensory inputs (what the monkeys were looking at) and top-down cognitive encoding inputs (what the monkeys were looking for).


Mechanisms underlying the varied mammary carcinogenicity of the environmental pollutant 6-nitrochrysene and its metabolites (-)-[R,R]- and (+)-[S,S]-1,2-dihydroxy-1,2-dihydro-6-nitrochrysene in the rat.
The mechanisms that can account for the remarkable mammary carcinogenicity of the environmental pollutant 6-nitrochrysene (6-NC) in the rat remain elusive. In our previous studies, we identified several 6-NC-derived DNA adducts in the rat mammary gland; one major adduct was derived from (±)-trans-1,2-dihydroxy-1,2-dihydro-6-nitrochrysene (1,2-DHD-6-NC). In the present study, we resolved the racemic (±)-1,2-DHD-6-NC into (-)-[R,R]- and (+)-[S,S]-1,2-DHD-6-NC and compared their in vivo mutagenicity and carcinogenicity in the mammary glands of female transgenic (BigBlue F344 × Sprague-Dawley)F1 rats harboring lacI/cII and Sprague-Dawley rats, respectively. Both [R,R]- and [S,S]-isomers exerted similar mutagenicity and carcinogenicity but were less potent than 6-NC. Additional in vivo and in vitro studies were then performed to explore possible mechanisms that can explain the higher potency of 6-NC than 1,2-DHD-6-NC. Using ELISA, we found that neither 6-NC nor 1,2-DHD-6-NC increased the levels of several inflammatory cytokines in plasma obtained from rats 24 h after treatment. In MCF-7 cells, as determined by immunoblotting, the effects of 6-NC and 1,2-DHD-6-NC on protein expression (p53, Akt, p38, JNK, c-myc, bcl-2, PCNA, and ERβ) were comparable; however, the expressions of AhR and ERα proteins were decreased by 6-NC but not 1,2-DHD-6-NC. The expression of both receptors was decreased in mammary tissues of rats treated with 6-NC. Our findings suggest that the differential effects of 6-NC and 1,2-DHD-6-NC on AhR and ERα could potentially account for the higher carcinogenicity of 6-NC in the rat mammary gland.


Structural Determinants for the Selectivity of the Positive KCa3.1 Gating Modulator 5-Methylnaphtho[2,1-d]oxazol-2-amine (SKA-121).
Intermediate-conductance (KCa3.1) and small-conductance (KCa2) calcium-activated K+ channels are gated by calcium binding to calmodulin (CaM) molecules associated with the calmodulin-binding domain (CaM-BD) of these channels. The existing KCa activators, such as naphtho[1,2-d]thiazol-2-ylamine (SKA-31), 6,7-dichloro-1H-indole-2,3-dione 3-oxime (NS309), and 1-ethylbenzimidazolin-2-one (EBIO), activate both channel types with similar potencies. In a previous chemistry effort, we optimized the benzothiazole pharmacophore of SKA-31 toward KCa3.1 selectivity and identified 5-methylnaphtho[2,1-d]oxazol-2-amine (SKA-121), which exhibits 40-fold selectivity for KCa3.1 over KCa2.3. To understand why introduction of a single CH3 group in five-position of the benzothiazole/oxazole system could achieve such a gain in selectivity for KCa3.1 over KCa2.3, we first localized the binding site of the benzothiazoles/oxazoles to the CaM-BD/CaM interface and then used computational modeling software to generate models of the KCa3.1 and KCa2.3 CaM-BD/CaM complexes with SKA-121. Based on a combination of mutagenesis and structural modeling, we suggest that all benzothiazole/oxazole-type KCa activators bind relatively "deep" in the CaM-BD/CaM interface and hydrogen bond with E54 on CaM. In KCa3.1, SKA-121 forms an additional hydrogen bond network with R362. In contrast, NS309 sits more "forward" and directly hydrogen bonds with R362 in KCa3.1. Mutating R362 to serine, the corresponding residue in KCa2.3 reduces the potency of SKA-121 by 7-fold, suggesting that R362 is responsible for the generally greater potency of KCa activators on KCa3.1. The increase in SKA-121's KCa3.1 selectivity compared with its parent, SKA-31, seems to be due to better overall shape complementarity and hydrophobic interactions with S372 and M368 on KCa3.1 and M72 on CaM at the KCa3.1-CaM-BD/CaM interface.


Molecular Face-Rotating Cube with Emergent Chiral and Fluorescence Properties.
Chiral cage compounds are mainly constructed from chiral precursors or based on the symmetry breaking during coordination-driven self-assembly. Herein, we present a strategy to construct chiral organic cages by restricting the P or M rotational configuration of tetraphenylethylene (TPE) faces through dynamic covalent chemistry. The combination of graph theory, experimental characterizations and theoretical calculations suggests emergent chirality of cages is originated from complex arrangements of TPE faces with different orientational and rotational configurations. Accompanied by the generation of chirality, strong fluorescence also emerged during cage formation, even in dilute solutions with various solvents. In addition, the circularly polarized luminescence of the cages is realized as a synergy of their dual chiral and fluorescence properties. Chirality and fluorescence of cages are remarkably stable, because intramolecular flipping of phenyl rings in TPE faces is restricted, as indicated by calculations. This study provides insight into construct chiral cages by the rational design through graph theory, and might facilitate further design of cages and other supramolecular assemblies from aggregation-induced emission active building blocks.


Application of microRNA in diagnosis and treatment of ovarian cancer.
Ovarian cancer has a poor prognosis because early detection is difficult and recurrent ovarian cancer is usually drug-resistant. The morbidity and mortality of ovarian cancer are high worldwide and new methods of diagnosis and therapy are needed. MicroRNAs (miRNAs) are posttranscriptional regulators of gene expression that are involved in carcinogenesis, metastasis, and invasion. Thus, miRNAs are likely to be useful as diagnostic and prognostic biomarkers and for cancer therapy. Many miRNAs have altered expression in ovarian cancer compared to normal ovarian tissues and these changes may be useful for diagnosis and treatment. For example, deficiencies of enzymes including Dicer and Drosha that are required for miRNA biogenesis may be adverse prognostic factors; miRNAs such as miR-214 and miR-31, which are involved in drug resistance, and the miR-200 family, which is implicated in metastasis, may serve as biomarkers; and transfection of downregulated miRNAs and inhibition of upregulated miRNAs may be effective for treatment of ovarian cancer. Chemotherapy targeting epigenetic mechanisms associated with miRNAs may also be effective to reverse gene silencing.


Detailed structural-functional analysis of the Krüppel-like factor 16 (KLF16) transcription factor reveals novel mechanisms for silencing Sp/KLF sites involved in metabolism and endocrinology.
Krüppel-like factor (KLF) proteins have elicited significant attention due to their emerging key role in metabolic and endocrine diseases. Here, we extend this knowledge through the biochemical characterization of KLF16, unveiling novel mechanisms regulating expression of genes involved in reproductive endocrinology. We found that KLF16 selectively binds three distinct KLF-binding sites (GC, CA, and BTE boxes). KLF16 also regulated the expression of several genes essential for metabolic and endocrine processes in sex steroid-sensitive uterine cells. Mechanistically, we determined that KLF16 possesses an activation domain that couples to histone acetyltransferase-mediated pathways, as well as a repression domain that interacts with the histone deacetylase chromatin-remodeling system via all three Sin3 isoforms, suggesting a higher level of plasticity in chromatin cofactor selection. Molecular modeling combined with molecular dynamic simulations of the Sin3a-KLF16 complex revealed important insights into how this interaction occurs at an atomic resolution level, predicting that phosphorylation of Tyr-10 may modulate KLF16 function. Phosphorylation of KLF16 was confirmed by in vivo (32)P incorporation and controlled by a Y10F site-directed mutant. Inhibition of Src-type tyrosine kinase signaling as well as the nonphosphorylatable Y10F mutation disrupted KLF16-mediated gene silencing, demonstrating that its function is regulatable rather than constitutive. Subcellular localization studies revealed that signal-induced nuclear translocation and euchromatic compartmentalization constitute an additional mechanism for regulating KLF16 function. Thus, this study lends insights on key biochemical mechanisms for regulating KLF sites involved in reproductive biology. These data also contribute to the new functional information that is applicable to understanding KLF16 and other highly related KLF proteins.


The oxygen-independent metabolism of cyclic monoterpenes in Castellaniella defragrans 65Phen.
The facultatively anaerobic betaproteobacterium Castellaniella defragrans 65Phen utilizes acyclic, monocyclic and bicyclic monoterpenes as sole carbon source under oxic as well as anoxic conditions. A biotransformation pathway of the acyclic β-myrcene required linalool dehydratase-isomerase as initial enzyme acting on the hydrocarbon. An in-frame deletion mutant did not use myrcene, but was able to grow on monocyclic monoterpenes. The genome sequence and a comparative proteome analysis together with a random transposon mutagenesis were conducted to identify genes involved in the monocyclic monoterpene metabolism. Metabolites accumulating in cultures of transposon and in-frame deletion mutants disclosed the degradation pathway. Castellaniella defragrans 65Phen oxidizes the monocyclic monoterpene limonene at the primary methyl group forming perillyl alcohol. The genome of 3.95 Mb contained a 70 kb genome island coding for over 50 proteins involved in the monoterpene metabolism. This island showed higher homology to genes of another monoterpene-mineralizing betaproteobacterium, Thauera terpenica 58EuT, than to genomes of the family Alcaligenaceae, which harbors the genus Castellaniella. A collection of 72 transposon mutants unable to grow on limonene contained 17 inactivated genes, with 46 mutants located in the two genes ctmAB (cyclic terpene metabolism). CtmA and ctmB were annotated as FAD-dependent oxidoreductases and clustered together with ctmE, a 2Fe-2S ferredoxin gene, and ctmF, coding for a NADH:ferredoxin oxidoreductase. Transposon mutants of ctmA, B or E did not grow aerobically or anaerobically on limonene, but on perillyl alcohol. The next steps in the pathway are catalyzed by the geraniol dehydrogenase GeoA and the geranial dehydrogenase GeoB, yielding perillic acid. Two transposon mutants had inactivated genes of the monoterpene ring cleavage (mrc) pathway. 2-Methylcitrate synthase and 2-methylcitrate dehydratase were also essential for the monoterpene metabolism but not for growth on acetate. The genome of Castellaniella defragrans 65Phen is related to other genomes of Alcaligenaceae, but contains a genomic island with genes of the monoterpene metabolism. Castellaniella defragrans 65Phen degrades limonene via a limonene dehydrogenase and the oxidation of perillyl alcohol. The initial oxidation at the primary methyl group is independent of molecular oxygen.


Molecular cloning of steroid 21-hydroxylase.
Congenital adrenal hyperplasia due to 21-hydroxylase (21-OH) deficiency is HLA-linked. The haplotype HLA-(A3);Bw47;DR7 is strongly associated with 21-OH deficiency and always carries a null allele at the complement C4A (Rodgers) locus. It seemed likely that this haplotype carries a deletion encompassing both the C4A and 21-OH loci. We hypothesized that the HLA-linked defect involved a structural gene for the adrenal microsomal cytochrome P-450 specific for steroid 21-hydroxylation. We isolated a plasmid with a 520 bp bovine adrenal cDNA insert encoding the middle third of the P-450 peptide. When human DNA was digested with Taq I restriction endonuclease and hybridized with the cDNA probe, DNA from 13 unrelated normal individuals yielded two hybridizing bands of equal intensity at 3.7 and 3.2 kb. The upper band was not present in DNA from a patient homozygous for Bw47. DNA from six unrelated patients heterozygous for Bw47 yielded, in five, diminished relative intensity of the upper band consistent with a heterozygous deletion, and complete disappearance of the upper band in one. Thus 21-OH deficiency sometimes results from the deletion of a gene and sometimes, presumably, from smaller mutations. This gene is probably located very near the C4A gene encoding the 4th component of complement.


Ca2+ release induced by cyclic ADP-ribose.
Cyclic ADP-ribose (cADPR) is the most potent Ca(2+)-mobilizing agent known. It has been found in many different cell types, where it is synthesized from its precursor NAD(+) by ADP-ribosyl cyclases. cADPR binds to Ca(2+) channels in the endoplasmic reticulum membrane to activate a Ca(2+)-release mechanism. This release is itself potentiated by elevated cytoplasmic Ca(2+) concentrations. Thus, cADPR may function as an endogenous regulator of Ca(2+)-induced Ca(2+) release, and there is excitement that it may also function as a Ca(2+)-mobilizing second messenger.


Effect of Sensory Deprivation on Maximal Force Abilities from Local to Non-local Digits.
The present study investigates the effect of sensory deprivation of the index and middle finger on motor function of all digits during maximal voluntary force production tasks. A total of 27 subjects performed maximal isometric pressing tasks by using different instructed finger combinations. Subjects completed the same tasks in two visits: a control visit when they had normal sensory feedback in all fingers, and an anesthesia visit when digital nerve blocks were performed on their right index and middle fingers. We evaluated three aspects of motor adaptation on both local (anesthetized) and non-local (non-anesthetized) digits during maximal force production: (1) task-relevant and overall force magnitude, (2) force directional application, and (3) digital individuation and force sharing. Our results indicate that selective digital anesthesia resulted in decreased maximal force magnitude, changed direction of force production, and significant changes extended to non-local digits. The motor weakness and inefficiency revealed in the non-local digits implies that sensory information from each digit can be shared across the digits to assist motor execution within the same hand.


Highly efficient gene transfer in naive human T cells with a murine leukemia virus-based vector.
Retroviral vectors based on the Moloney murine leukemia virus (MuLV) have become the primary tool for gene delivery into hematopoietic cells, but clinical trials have been hampered by low transduction efficiencies. Recently, we and others have shown that gene transfer of MuLV-based vectors into T cells can be significantly augmented using a fibronectin-facilitated protocol. Nevertheless, the relative abilities of naive (CD45RA(+)) and memory (CD45RO(+)) lymphocyte subsets to be transduced has not been assessed. Although naive T cells demonstrate a restricted cytokine profile following antigen stimulation and a decreased susceptibility to infection with human immunodeficiency virus, it was not clear whether they could be efficiently infected with a MuLV vector. This study describes conditions that permitted gene transfer of an enhanced green fluorescent protein-expressing retroviral vector in more than 50% of naive umbilical cord (UC) blood and peripheral blood (PB) T cells following CD3/CD28 ligation. Moreover, treatment of naive T cells with interleukin-7 resulted in the maintenance of a CD45RA phenotype and gene transfer levels approached 20%. Finally, it was determined that parameters for optimal transduction of CD45RA(+) T cells isolated from PB and UC blood differed: transduction of the UC cells was significantly increased by the presence of autologous mononuclear cells (24.5% versus 56.5%). Because naive T cells harbor a receptor repertoire that allows them to respond to novel antigens, the development of protocols targeting their transduction is crucial for gene therapy applications. This approach will also allow the functions of exogenous genes to be evaluated in primary nontransformed naive T cells.


IAPAC Guidelines for Optimizing the HIV Care Continuum for Adults and Adolescents.
An estimated 50% of people living with HIV (PLHIV) globally are unaware of their status. Among those who know their HIV status, many do not receive antiretroviral therapy (ART) in a timely manner, fail to remain engaged in care, or do not achieve sustained viral suppression. Barriers across the HIV care continuum prevent PLHIV from achieving the therapeutic and preventive effects of ART. A systematic literature search was conducted, and 6132 articles, including randomized controlled trials, observational studies with or without comparators, cross-sectional studies, and descriptive documents, met the inclusion criteria. Of these, 1047 articles were used to generate 36 recommendations to optimize the HIV care continuum for adults and adolescents. Recommendations are provided for interventions to optimize the HIV care environment; increase HIV testing and linkage to care, treatment coverage, retention in care, and viral suppression; and monitor the HIV care continuum.


Quantitative trait loci for resistance to fusarium head blight in a Chinese wheat landrace Haiyanzhong.
Fusarium head blight (FHB) of wheat causes not only significant reduction in grain yield and end-use quality, but also the contamination of the grain with mycotoxins that are detrimental to human and animal health after consumption of infected grain. Growing resistant varieties is an effective approach to minimize the FHB damage. The Chinese wheat landrace Haiyanzhong (HYZ) shows a high level of resistance to FHB. To identify quantitative trait loci (QTL) that contribute to FHB resistance in HYZ, 136 recombinant inbred lines (RIL) were developed from a cross of HYZ and Wheaton, a hard spring wheat cultivar from the USA. The RIL and their parents were evaluated for percentage of scabbed spikelets (PSS) in both greenhouse and field environments. Five QTL were detected for FHB resistance in HYZ with one major QTL on 7DL. The 7DL QTL peaked at SSR marker Xwmc121, which is flanked by the SSR markers Xcfd46 and Xwmc702. This QTL explained 20.4-22.6% of the phenotypic variance in individual greenhouse experiments and 15.9% in a field experiment. Four other minor QTL on 6BS (two QTL), 5AS and 1AS each explained less than 10% of the phenotypic variance in individual experiments. HYZ carried the favorable alleles associated with FHB resistance at the QTL on 7DL, 6BS and 5AS, and the unfavorable allele at the QTL on 1AS. The major QTL on 7D can be used to improve the FHB resistance in wheat breeding programs and add diversity to the FHB resistance gene pool.


Helicoverpa armigera preference and performance on three cultivars of short-duration pigeonpea (Cajanus cajan): the importance of whole plant assays.
Helicoverpa armigera is a major pest of pigeonpea (Cajanus cajan). Efforts to develop pigeonpea varieties resistant to H. armigera attack have been met with limited success, despite reports of high levels of resistance to H. armigera in wild relatives of pigeonpea and reports of low to moderate levels of resistance in cultivated varieties. Here we examined H. armigera oviposition preference and larval performance on whole plants of three cultivars of short-duration pigeonpea: a susceptible control (ICPL 87) and two cultivars with purported host-plant resistance (ICPL 86012 and ICPL 88039). In our no-choice oviposition experiment, H. armigera laid similar numbers of eggs on all three cultivars tested, but under choice conditions moths laid slightly more eggs on ICPL 88039. Larval growth and development were affected by cultivar, and larvae grew to the largest size (weight) and developed fastest on ICPL 86012. Moths laid most of their eggs on floral structures, sites where subsequent early instar larvae overwhelmingly fed. Experimentally placing neonate larvae at different locations on plants demonstrated that larvae placed on flowers experienced greater survival, faster development, and greater weight gain than those placed on leaves. The type and density of trichomes (a potential resistance trait) differed among cultivars and plant structures, but larvae selected to feed at sites where trichomes were absent. Future work examining host-plant resistance against H. armigera should incorporate the behavioural preference of moths and larvae in experiments using whole plants as opposed to bioassays of excised plant parts in Petri dishes. © 2022 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.


Volunteer Impact on Health-Related Outcomes for Seniors: a Systematic Review And Meta-Analysis.
Volunteers are increasingly promoted to improve health-related outcomes for community-dwelling elderly without synthesized evidence for effectiveness. This systematic review and meta-analysis evaluates the effects of unpaid volunteer interventions on health-related outcomes for such seniors. MEDLINE, EMBASE and Cochrane (CENTRAL) were searched up to November 2018. We included English language, randomized trials. Two reviewers independently identified studies, extracted data, and assessed evidence certainty (using GRADE). Meta-analysis used random-effects models. Univariate meta-regressions investigated the relationship between volunteer intervention effects and trial participant age, percentage females, and risk of bias. 28 included studies focussed on seniors with a variety of chronic conditions (e.g., dementia, diabetes) and health states (e.g., frail, palliative). Volunteers provided a range of roles (e.g., counsellors, educators and coaches). Low certainty evidence found that volunteers may improve both physical function (MD = 3.2 points on the 100-point SF-36 physical component score [PCS]; 95% CI: 1.09, 5.27) and physical activity levels (SMD = 0.5, 95% CI: 0.14 to 0.83). Adverse events were not increased. Volunteers may increase physical activity levels and subjective ratings of physical function for seniors without apparent harm. These findings support the WHO call to action on evidence-based policies to align health systems in support of older adults.


How much margin do we need for pelvic lymph nodes irradiation in the era of IGRT?
Background and purpose: Image guided radiotherapy (IGRT) without 6 degree of freedom couch can only correct the translational setup errors of pelvic radiotherapy. But errors introduced by rotation and deformation of CTV can't be adjusted in most of IGRT systems. This article is to evaluate these errors and to provide recommendations on the margin needed in the era of IGRT. Material and methods: 218 patients who received pelvic radiotherapy in PUMC Hospital from 2012 to 2014 were included. A simulation CT and a CBCT were acquired for every patient. 3D and 6D registrations of CT and CBCT were applied. 9 bony landmarks were marked and distances of each landmark between CT and CBCT were measured in three directions. Results: Without image guidance, movements of landmarks in the directions of LR, AP and SI were 0.4 ± 2.5 mm, 1.3 ± 3.8 mm and 1.5 ± 5.0 mm respectively, with 3D-registration, movements were 0.0 ± 1.5 mm, 0.7± 2.8 mm and 0.6± 3.2 mm, and with 6D-registration, movements were 0.0 ± 0.5 mm, 0.2 ± 1.0 mm and 0.2 ± 1.1 mm in each direction. Conclusions: IGRT could reduce setup errors. IGRT with 6D treatment couches could further reduce setup errors compared to 3D couches. For centers without IGRT, we suggest CTV-PTV margins of 6 mm, 9 mm and 12 mm in LR, AP and SI directions respectively, margins of 3 mm, 6.5 mm and 7 mm for the use of daily IGRT with 3D couch and 2 mm, 3 mm and 3 mm for 6D couch.


Treatment of nitrogen-rich wastewater using partial nitrification and anammox in the CANON process.
Partial nitrification combined with Anammox in a single reactor (the CANON process) is an energy-efficient N-removal technology that could substantially lower the N-load of a WWTP by separate treatment of nitrogen-rich side streams, preventing the need for extensive expansion and reducing the total energy requirement. This study looks at the enrichment of Anammox from activated sludge and its application in the CANON process on lab-scale. The aim was to identify the critical process control parameters necessary for successful operation of CANON. An Anammox culture capable of removing 0.6 kg N/m3/d was enriched in 14 weeks in a sequencing batch reactor. Nitrifying biomass was inoculated into the Anammox reactor (10% v/v) together with limited oxygen supply (< 8 mL/min) to initiate the CANON process in continuous culture. The small flocs formed by the biomass (< 1000 microm) were sensitive to low O2 concentrations (< 0.1 mg/L) which prevented simultaneous nitrification and Anammox. Operation with 20 min aerobiosis and 30 min anaerobiosis was necessary to achieve sustained, completely autotrophic N-removal for an extended period at a rate of 0.08 kg N/m3/d. Essential process control parameters for stable CANON operation were the nitrite concentration, oxygen concentration, pH and the temperature.


Open surgical wounds: how does Aquacel compare with wet-to-dry gauze?
To compare the healing rates of a hydrofibre dressing (Aquacel) and normal wet-to-dry gauze in the treatment of open surgical wounds. Fifty patients with open surgical wounds were randomized to receive either saline-moistened gauze or Aquacel. The rate of wound healing was measured as ml/day (deep wounds) or cm2/day (superficial wounds) at each dressing change until an investigator blinded to the patient group diagnosed the wounds as having healed or the patient was withdrawn from the study. Of the 50 patients, seven were withdrawn from the study after the first evaluation. Of the remaining 43 patients, 21 had been randomly allocated to the gauze group and 22 to the Aquacel group. For deep wounds, a mean change in the wound healing rate of 1.9 +/- 1.3 cm3/day was reported for the gauze group and 2.9 +/- 2.3 cm3/day for the Aquacel group. These results approach statistical significance (p = 0.082). For superficial wounds, the mean change in the healing rate was 1.6 +/- 1.5 cm2/day for the gauze group and 1.9 +/- 2.2 cm2/day for the Aquacel group, but this was not statistically significant. Aquacel appears to be at least as effective as wet-to-dry gauze in the healing of open surgical wounds.


Developmental expression of potassium-channel subunit Kv3.2 within subpopulations of mouse hippocampal inhibitory interneurons.
The developmental expression of the voltage-gated potassium channel subunit, Kv3.2, and its localization within specific mouse hippocampal inhibitory interneuron populations were determined using immunoblotting and immunohistochemical techniques. Using immunoblotting techniques, the Kv3.2 protein was weakly detected at postnatal age day 7 (P7), and full expression was attained at P21 in tissue extracts from homogenized hippocampal preparations. A similar developmental profile was observed using immunohistochemical techniques in hippocampal tissue sections. Kv3.2 protein expression was clustered on the somata and proximal dendrites of presumed inhibitory interneurons. Using double immunofluorescence, Kv3.2 subunit expression was detected on subpopulations of GABAergic inhibitory interneurons. Kv3.2 was detected in approximately 100% of parvalbumin-positive interneurons, 86% of interneurons expressing nitric oxide synthase, and approximately 50% of somatostatin-immunoreactive cells. Kv3.2 expression was absent from both calbindin- and calretinin-containing interneurons. Using immunoprecipitation, we further demonstrate that Kv3.2 and its related subunit Kv3.1b are coexpressed within the same protein complexes in the hippocampus. These data demonstrate that potassium channel subunit Kv3.2 expression is developmentally regulated in a specific set of interneurons. The vast majority of these interneuron subpopulations possess a "fast-spiking" phenotype, consistent with a role for currents through Kv3.2 containing channels in determining action potential kinetics in these cells.


Selective serotonin reuptake inhibitor drug interactions in patients receiving statins.
Elderly patients commonly receive statin drugs for the primary or secondary prevention of cardiovascular and cerebrovascular events. Elderly patients also commonly receive antidepressant drugs, usually selective serotonin reuptake inhibitors (SSRIs), for the treatment of depression, anxiety, or other conditions. SSRIs are associated with many pharmacokinetic drug interactions related to the inhibition of the cytochrome P450 (CYP) metabolic pathways. There is concern that drugs that inhibit statin metabolism can trigger statin adverse effects, especially myopathy (which can be potentially serious, if rhabdomyolysis occurs). However, a detailed literature review of statin metabolism and of SSRI effects on CYP enzymes suggests that escitalopram, citalopram, and paroxetine are almost certain to be safe with all statins, and rosuvastatin, pitavastatin, and pravastatin are almost certain to be safe with all SSRIs. Even though other SSRI-statin combinations may theoretically be associated with risks, the magnitude of the pharmacokinetic interaction is likely to be below the threshold for clinical significance. Risk, if at all, lies in combining fluvoxamine with atorvastatin, simvastatin, or lovastatin, and even this risk can be minimized by using lower statin doses and monitoring the patient.


DNA content and regional metastases in patients with advanced laryngeal squamous carcinoma. Department of Veterans Affairs Laryngeal Study Group.
Abnormal DNA content of cancer cells in a primary tumor is thought to reflect the altered proliferative capacity of that cancer and overall patient prognosis. Recently, increased DNA content has been associated with early tumor recurrence in patients with advanced laryngeal cancer. To determine if DNA content correlates with a tumor's metastatic behavior, 94 previously untreated patients with stage III (n = 54) or stage IV (n = 40) squamous carcinoma of the larynx were prospectively studied. The adjusted DNA index (aDI) of the primary tumor was measured by computerized cytomorphometry, and correlations with regional metastases, number of positive nodes, extracapsular spread, and pattern of relapse were determined. Median follow-up was 35 months. Of 94 patients, 73 underwent neck dissection. Positive nodes were found in 51 patients (70%) and three or more positive nodes were found in 31 patients (42%). High aDI (> .024) was significantly associated with the presence and number of histologically positive nodes (P = .016). Among patients with high aDI, 19 (50%) of 38 had three or more nodes positive compared to 12 (21%) of 56 of those with low aDI. Median aDI levels in patients with three or more nodes were significantly higher than in patients with 1 or 2 nodes (P = .003). Time to recurrence was shorter in patients with high aDI (P = .0078). These findings provide further evidence that increased DNA content is associated with more aggressive laryngeal cancers having high rates of multiple lymph node metastases and worse overall prognosis.


The prevalence of superior vena cava anomalies as detected in cardiac implantable electronic device recipients at a tertiary cardiology centre over a 12-year period.
The vast majority of cardiac implantable electronic device (CIED) recipients require transvenous lead insertion, which may be hindered by the presence of venous anomalies. The aim of this study was to determine the prevalence and variations of persistent left superior vena cava (PLSVC) and to conduct subsequent outpatient follow-up in terms of device function and the clinical condition of the recipients using data from CIED placement procedures conducted over a 12-year period. The study population included patients undergoing first-time transvenous implantation of cardiac pacemakers and implantable cardioverter-defibrillators (ICDs). The presence of PLSVC was determined based on intra-procedure venography. Outpatient follow-up involved assessments of patient condition, radiological imaging, and CIED function. Of a total of 4708 CIED recipients, PLSVC was detected in eight patients (mean age 65.5±13.9); five of them had double superior vena cava (DSVC), including three cases in which the vessels were bridged with a brachiocephalic vein (BCV). Three patients presented PLSVC associated with the absence of the right superior vena cava (RSVC), a very rare anomaly. Seven patients remain under observation, for a total of 78.4±48.4 months of follow-up. The rate of venous anomalies in the form of PLSVC detected in the evaluated population was 0.17%. These PLSVC cases were asymptomatic, which hindered their earlier detection. The presence of these anomalies made the procedures more challenging for the operator and increased the perioperative complication rates; however, neither patient condition nor CIED function was affected based on the long-term outpatient follow-up.


Towards chemical accuracy for the thermodynamics of large molecules: new hybrid density functionals including non-local correlation effects.
Two hybrid density functionals that include a second-order perturbation correction for non-local correlation effects are tested for the full G3/05 test set. Very large AO basis sets including core-polarization/correlation functions have been employed that yield for the first time results quite close to the basis set limit for this set. The B2-PLYP functional and the new mPW2-PLYP approach with a modified exchange part give by far the lowest MAD over the whole G3/05 set ever reported for a DFT method (2.5 and 2.1 kcal mol(-1), respectively). The big improvement compared to common density functionals is further demonstrated by the reduction of the maximum and minimum errors (outliers) and by much smaller errors for complicated molecular systems.


Promoting healthy weight in primary school children through physical activity and nutrition education: a pragmatic evaluation of the CHANGE! randomised intervention study.
This pragmatic evaluation investigated the effectiveness of the Children's Health, Activity and Nutrition: Get Educated! (CHANGE!) Project, a cluster randomised intervention to promote healthy weight using an educational focus on physical activity and healthy eating. Participants (n = 318, aged 10-11 years) from 6 Intervention and 6 Comparison schools took part in the 20 weeks intervention between November 2010 and March/April 2011. This consisted of a teacher-led curriculum, learning resources, and homework tasks. Primary outcome measures were waist circumference, body mass index (BMI), and BMI z-scores. Secondary outcomes were objectively-assessed physical activity and sedentary time, and food intake. Outcomes were assessed at baseline, at post-intervention (20 weeks), and at follow-up (30 weeks). Data were analysed using 2-level multi-level modelling (levels: school, student) and adjusted for baseline values of the outcomes and potential confounders. Differences in intervention effect by subgroup (sex, weight status, socio-economic status) were explored using statistical interaction. Significant between-group effects were observed for waist circumference at post-intervention (β for intervention effect =-1.63 (95% CI = -2.20, -1.07) cm, p<0.001) and for BMI z-score at follow-up (β=-0.24 (95% CI = -0.48, -0.003), p=0.04). At follow-up there was also a significant intervention effect for light intensity physical activity (β=25.97 (95% CI = 8.04, 43.89) min, p=0.01). Interaction analyses revealed that the intervention was most effective for overweight/obese participants (waist circumference: β=-2.82 (95% CI = -4.06, -1.58) cm, p<0.001), girls (BMI: β=-0.39 (95% CI = -0.81, 0.03) kg/m2, p=0.07), and participants with higher family socioeconomic status (breakfast consumption: β=8.82 (95% CI = 6.47, 11.16), p=0.07). The CHANGE! intervention positively influenced body size outcomes and light physical activity, and most effectively influenced body size outcomes among overweight and obese children and girls. The findings add support for the effectiveness of combined school-based physical activity and nutrition interventions. Additional work is required to test intervention fidelity and the sustained effectiveness of this intervention in the medium and long term. Current Controlled Trials ISRCTN03863885.


Mechanism of leg stiffness adjustment for hopping on surfaces of different stiffnesses.
When humans hop in place or run forward, leg stiffness is increased to offset reductions in surface stiffness, allowing the global kinematics and mechanics to remain the same on all surfaces. The purpose of the present study was to determine the mechanism for adjusting leg stiffness. Seven subjects hopped in place on surfaces of different stiffnesses (23-35,000 kN/m) while force platform, kinematic, and electromyographic data were collected. Leg stiffness approximately doubled between the most stiff surface and the least stiff surface. Over the same range of surfaces, ankle torsional stiffness increased 1.75-fold, and the knee became more extended at the time of touchdown (2.81 vs. 2.65 rad). We used a computer simulation to examine the sensitivity of leg stiffness to the observed changes in ankle stiffness and touchdown knee angle. Our model consisted of four segments (foot, shank, thigh, head-arms-trunk) interconnected by three torsional springs (ankle, knee, hip). In the model, an increase in ankle stiffness 1.75-fold caused leg stiffness to increase 1.7-fold. A change in touchdown knee angle as observed in the subjects caused leg stiffness to increase 1.3-fold. Thus both joint stiffness and limb geometry adjustments are important in adjusting leg stiffness to allow similar hopping on different surfaces.


Differences in the anatomic distribution of immediate-early gene expression in amygdala and angular bundle kindling development.
Kindling is a model in which fleeting changes of neuronal activity produce a lifelong modification of neuronal structure and function in the mature nervous system. Immediate-early genes (IEGs) such as c-fos have been implicated as a causal link in the chain of molecular events coupling fleeting pathologic activity to lasting hyperexcitability. Identification of the brain structures exhibiting IEG expression during the evolution of kindling is necessary to guide investigations of the phenotypic consequences. We used in situ hybridization histochemistry to identify the structures exhibiting expression of multiple IEGs during the evolution of amygdala kindling and compared this to the pattern following angular bundle kindling. The principal findings included that: (1) generalized limbic and clonic motor (class 5) kindled seizures evoked by stimulation of one amygdala induced the expression of IEGs in a small subset of limbic structures with remarkable symmetry between the two hemispheres; (2) the anatomic extent of seizure-evoked expression of c-fos mRNA expanded progressively following focal limbic and motor (classes 0-3) seizures during the development of amygdala kindling; c-fos mRNA was detected first ipsilaterally in AM, ACO, and PC and with higher-class seizures in hippocampal formation and homologous structures contralaterally, and (3) class 5 seizures evoked by stimulation of two different sites in the limbic system (amygdala or angular bundle) induced IEG expression in distinct but partially overlapping anatomic structures. We propose that synaptic activation of glutamate receptors contributes to the expression of these diverse IEGs throughout the forebrain. The findings provide a constellation of anatomic structures in which to investigate the structural and functional consequences of IEG expression.


Competing risks of mortality in prostate cancer.
We have observed that in prostate cancer older age, black race, high stage and no treatment are independently associated with higher disease specific and overall mortality. Because disease nonspecific mortality was not examined one cannot reasonably infer that higher overall mortality is largely due to higher disease specific mortality. To understand better the interactions of independent prognostic variables with overall mortality we jointly evaluated their effects on type of death and time to death. Using the prostate cancer patients of the 1973 to 1990 public use tape of the Surveillance, Epidemiology and End Results program, we performed competing risks analysis with multivariate accelerated failure time model to examine if the prognostic factors were associated with type of death and time to death. Older age, black race and no treatment were independently associated with higher relative risk of disease specific and nonspecific mortality. Localized stage was associated with lower and higher disease specific and nonspecific mortality than regional stage, respectively. The relative risks of disease specific and nonspecific mortality were significantly different from each other for all prognostic factors. To our knowledge, a competing risks analysis in prostate cancer has not been done previously. While all prognostic factors were associated with disease specific and nonspecific mortality, the relative contribution of disease specific and nonspecific mortality to overall mortality varied by prognostic factor, with some prognostic factors associated with relatively high disease specific mortality while others were associated with relatively high disease nonspecific mortality. The similarity of the associations of age, race and treatment with disease specific and nonspecific mortality suggests that they mark an unidentified factor(s) that affects patient health nonspecifically. Such a factor(s) may partly or completely explain the association of treatment with lower disease specific mortality.


Understanding tremor in multiple sclerosis: prevalence, pathological anatomy, and pharmacological and surgical approaches to treatment.
Given that tremor is one of the most prevalent and disabling features of multiple sclerosis (MS), we will review the most significant milestones in tremor in this disease in recent years, focusing on prevalence, clinical features, anatomical basis, and treatment. Data for this review were identified by searching MEDLINE with the search terms "multiple sclerosis" and "tremor". References were also identified from relevant articles published between January 1966 and May 2012. The predominant type of MS tremor is a large-amplitude, postural, and kinetic tremor, which most commonly affects the arms, although tremor can also involve head, neck, vocal cords, and trunk. Involvement of the tongue, jaw, or palate has not been reported. Although the anatomical basis underlying tremor in MS is poorly understood, the link between the cerebellum and the MS-related tremor is supported by clinical and experimental studies. Currently available medication is often unsuccessful in most cases. Surgical treatment can be a satisfactory alternative to treat severe and disabling tremor. Tremor in MS patients could be considered as an advanced consequence of the disease and its presence suggests a more aggressive course. MS tremor can be severe and very disabling for a small group of patients. Treatment of MS tremor remains a great challenge. Recent studies suggest that dissociating tremor from cerebellar dysfunction using selected clinical tests would be the key issue to successful surgical treatment. Understanding the pathophysiology and biochemistry of tremor production in MS may lead to new therapeutic approaches.


An assessment of set up position for MRI scanning for the purposes of rectal cancer radiotherapy treatment planning.
A magnetic resonance (MR) scanner for radiotherapy treatment simulation was commissioned in our department in June 2013. Practical set up and MR image quality trade-offs using a variety of patient positions and immobilisation devices routinely used in the treatment planning of rectal cancer patients were considered. The study also aimed to investigate the MR compatibility of the device materials with a focus on temperature changes during routine clinical examinations. Ten volunteers were scanned: (1) Prone on a Civco Contoura Bellyboard (BBB), (2) Prone on a Civco MR Series Bellyboard (WBB), (3) Prone with no bellyboard and (4) Supine. All scans were performed with a T2 weighted (T2 -w) turbo spin echo (TSE) sequence. Images were scored by five assessors for: (1) ease of identifying specific organs, (2) overall image quality and (3) signal to noise ratio (SNR). Temperature changes were measured for each volunteer in each position. Both expert scores and SNR analysis demonstrated that images obtained in the supine position allowed for easier and clearer delineation of the organs. Image factors such as artefacts and noise, along with the overall image quality, also performed better in the supine position. The carbon fibre bellyboard did not demonstrate significant heating during scanning with the T2 -w TSE transverse sequence. A supine position was determined to be superior to the other positions in a majority of comparisons. The volunteers did not experience any increased temperature changes during scanning on the bellyboard in comparison to the other positions.


Association between oral tori, occlusal force, and mandibular cortical index.
To assess possible associations between torus palatinus (TP), torus mandibularis (TM), occlusal force (OF), Klemetti's classes, mandibular cortical index (MCI), and sociodemographic variables in a selected sample of Jordanians. Previous studies have suggested that oral tori are benign anatomical variations probably related to several factors such as functional stress, gender predisposition, number of teeth present, and nutritional factors. A total of 130 Jordanian adults were included. Shape, location, and appearance of tori were recorded from study casts. Size was measured using digital calipers. OF was recorded in Newtons. MCI and Klemetti's classification were assessed using panoramic radiographs. Data were statistically analyzed, and level of significance was set at P < .05. The mean ± SD recorded OF was 524 ± 183 N in the tori group, compared to 417 ± 172 N in controls (P = .001). OF was significantly higher in males compared to females (P < .001), and was significantly higher in subjects with TM only (543 ± 201 N) (P < .001). TP and TM were predominantly more than 6 mm in size. The average OF in subjects with Klemetti class 3 was 418 ± 174 N, while OF in Klemetti class 1 subjects amounted to 535 ± 187 N (P = .043). Average OF was significantly higher in tori subjects and in males (if compared to controls and female subjects, respectively). The presence of oral tori, Klemetti's classification, and MCI ratio seems to be positively correlated with OF.


Characterization of 14C in Swedish light water reactors.
This paper presents the results of a 4-y investigation of 14C in different waste streams of both boiling water reactors (BWRs) and pressurized water reactors (PWRs). Due to the potential impact of 14C on human health, minimizing waste and releases from the nuclear power industry is of considerable interest. The experimental data and conclusions may be implemented to select appropriate waste management strategies and practices at reactor units and disposal facilities. Organic and inorganic 14C in spent ion exchange resins, process water systems, ejector off-gas and replaced steam generator tubes were analyzed using a recently developed extraction method. Separate analysis of the chemical species is of importance in order to model and predict the fate of 14C within process systems as well as in dose calculations for disposal facilities. By combining the results of this investigation with newly calculated production rates, mass balance assessments were made of the 14C originating from production in the coolant. Of the 14C formed in the coolant of BWRs, 0.6-0.8% was found to be accumulated in the ion exchange resins (core-specific production rate in the coolant of a 2,500 MWth BWR calculated to be 580 GBq GW(e)(-1) y(-1)). The corresponding value for PWRs was 6-10% (production rate in a 2,775 MWth PWR calculated to be 350 GBq GW(e)(-1) y(-1)). The 14C released with liquid discharges was found to be insignificant, constituting less than 0.5% of the production in the coolant. The stack releases, routinely measured at the power plants, were found to correspond to 60-155% of the calculated coolant production, with large variations between the BWR units.


Polymorphonuclear leukocytes as a significant source of tumor necrosis factor-alpha in endotoxin-challenged lung tissue.
The kinetic expression and potential cellular source of tumor necrosis factor-alpha (TNF-alpha) in lipopolysaccharide-(LPS) induced acute lung inflammation was investigated using a rat model by Northern blot analysis, in situ hybridization and immunohistochemistry. LPS induced a polymorphonuclear leukocyte infiltrate in the lung that peaked between 6 and 24 hours. TNF-alpha messenger (m)RNA was strongly induced by LPS in whole lung tissues shown by Northern analysis. Both alveolar macrophages and polymorphonuclear leukocytes (PMNs), purified from bronchoalveolar lavage fluids of LPS-treated rats, were shown to express TNF-alpha mRNA by Northern analysis. However, PMNs displayed several times more TNF-alpha mRNA, relative to actin mRNA, than alveolar macrophages at 6 and 12 hours. By in situ hybridization, most of the cells positive for TNF-alpha mRNA at 6 and 12 hours seemed to be PMNs located within the tissue near bronchioles or vessels. By immunohistochemistry, TNF-alpha protein was localized mainly to alveolar macrophages at early times (1 to 3 hours) after LPS challenge, and thereafter, PMNs seemed to be the predominant source of TNF-alpha protein as more than 90% of total intraalveolar positive cells at 6 and 12 hours were PMN. Thus, our data provide the first in vivo evidence that PMNs can serve as a significant source of TNF-alpha at sites of acute inflammation.


Distribution of Kv1-like potassium channels in the electromotor and electrosensory systems of the weakly electric fish Apteronotus leptorhynchus.
The electromotor and electrosensory systems of the weakly electric fish Apteronotus leptorhynchus are model systems for studying mechanisms of high-frequency motor pattern generation and sensory processing. Voltage-dependent ionic currents, including low-threshold potassium currents, influence excitability of neurons in these circuits and thereby regulate motor output and sensory filtering. Although Kv1-like potassium channels are likely to carry low-threshold potassium currents in electromotor and electrosensory neurons, the distribution of Kv1 alpha subunits in A. leptorhynchus is unknown. In this study, we used immunohistochemistry with six different antibodies raised against specific mammalian Kv1 alpha subunits (Kv1.1-Kv1.6) to characterize the distribution of Kv1-like channels in electromotor and electrosensory structures. Each Kv1 antibody labeled a distinct subset of neurons, fibers, and/or dendrites in electromotor and electrosensory nuclei. Kv1-like immunoreactivity in the electrosensory lateral line lobe (ELL) and pacemaker nucleus are particularly relevant in light of previous studies suggesting that potassium currents carried by Kv1 channels regulate neuronal excitability in these regions. Immunoreactivity of pyramidal cells in the ELL with several Kv1 antibodies is consistent with Kv1 channels carrying low-threshold outward currents that regulate spike waveform in these cells (Fernandez et al., J Neurosci 2005;25:363-371). Similarly, Kv1-like immunoreactivity in the pacemaker nucleus is consistent with a role of Kv1 channels in spontaneous high-frequency firing in pacemaker neurons. Robust Kv1-like immunoreactivity in several other structures, including the dorsal torus semicircularis, tuberous electroreceptors, and the electric organ, indicates that Kv1 channels are broadly expressed and are likely to contribute significantly to generating the electric organ discharge and processing electrosensory inputs.


Developments in influenza vaccination coverage in Italy over five seasons (2001-2006).
This study aims at assessing trends in influenza vaccination coverage from 2001 to 2006 in Italy, at understanding the drivers and barriers to vaccination, and to identify vaccination intentions for season 2006/07. We conducted telephone-based household surveys representative of the population from age 14. Total sample size was 10,026 persons. The influenza vaccination coverage rate in Italy increased over the years, reaching 24.1% in season 2005/06. In the elderly (> 65), the vaccination rate reached 63.4% in 2005/06. The increase between seasons 2004/05 and 2005/06 was statistically significant. A recommendation for the doctor/nurse, considering influenza as a serious illness, older age and not being in very good health were the most frequent reasons for getting vaccinated. Four percent of those vaccinated in 2005/06 indicated the threat of avian flu as a reason. For the winter 2006/07, 30% of the population intended to get vaccinated, indicating the potential to increase vaccination coverage next year. The threat of avian influenza may have contributed to a substantial recent increase in season 2005/06 as compared to the year before. Despite of this positive evolution, major effort is still needed to reach WHO objectives for 2010 (75% vaccination coverage rate in the elderly population).


Global access of patients with kidney disease to health technologies and medications: findings from the Global Kidney Health Atlas project.
Access to essential medications and health products is critical to effective management of kidney disease. Using data from the ISN Global Kidney Health Atlas multinational cross-sectional survey, global access of patients with kidney disease to essential medications and health products was examined. Overall, 125 countries participated, with 118 countries, composing 91.5% of the world's population, providing data on this domain. Most countries were unable to access eGFR and albuminuria in their primary care settings. Only one-third of low-income countries (LICs) were able to measure serum creatinine and none were able to access eGFR or quantify proteinuria. The ability to monitor diabetes mellitus through serum glucose and glycated hemoglobin measurements was suboptimal. Pathology services were rarely available in tertiary care in LICs (12%) and lower middle-income countries (45%). While acute and chronic hemodialysis services were available in almost all countries, acute and chronic peritoneal dialysis services were rarely available in LICs (18% and 29%, respectively). Kidney transplantation was available in 79% of countries overall and in 12% of LICs. While over one-half of all countries publicly funded RRT and kidney medications with or without copayment, this was less common in LICs and lower middle-income countries. In conclusion, this study demonstrated significant gaps in services for kidney care and funding that were most apparent in LICs and lower middle-income countries.


Transcriptional robustness and protein interactions are associated in yeast.
Robustness to insults, both external and internal, is a characteristic feature of life. One level of biological organization for which noise and robustness have been extensively studied is gene expression. Cells have a variety of mechanisms for buffering noise in gene expression, but it is not completely clear what rules govern whether or not a given gene uses such tools to maintain appropriate expression. Here, we show a general association between the degree to which yeast cells have evolved mechanisms to buffer changes in gene expression and whether they possess protein-protein interactions. We argue that this effect bears an affinity to epistasis, because yeast appears to have evolved regulatory mechanisms such that distant changes in gene copy number for a protein-protein interaction partner gene can alter a gene's expression. This association is not unexpected given recent work linking epistasis and the deleterious effects of changes in gene dosage (i.e., the dosage balance hypothesis). Using gene expression data from artificial aneuploid strains of bakers' yeast, we found that genes coding for proteins that physically interact with other proteins show less expression variation in response to aneuploidy than do other genes. This effect is even more pronounced for genes whose products interact with proteins encoded on aneuploid chromosomes. We further found that genes targeted by transcription factors encoded on aneuploid chromosomes were more likely to change in expression after aneuploidy. We suggest that these observations can be best understood as resulting from the higher fitness cost of misexpression in epistatic genes and a commensurate greater regulatory control of them.


Flies evade looming targets by executing rapid visually directed banked turns.
Avoiding predators is an essential behavior in which animals must quickly transform sensory cues into evasive actions. Sensory reflexes are particularly fast in flying insects such as flies, but the means by which they evade aerial predators is not known. Using high-speed videography and automated tracking of flies in combination with aerodynamic measurements on flapping robots, we show that flying flies react to looming stimuli with directed banked turns. The maneuver consists of a rapid body rotation followed immediately by an active counter-rotation and is enacted by remarkably subtle changes in wing motion. These evasive maneuvers of flies are substantially faster than steering maneuvers measured previously and indicate the existence of sensory-motor circuitry that can reorient the fly's flight path within a few wingbeats.


Characterization of chicken natural resistance-associated macrophage protein encoding genes (Nramp1 and Nramp2) and association with salmonellosis resistance.
Natural resistance-associated macrophage protein 1 and 2 encoding genes (Nramp1 and Nramp2) are related to many diseases. We cloned the cDNA of chicken Nramp1 and Nramp2 genes, characterized their expression and polymorphisms, and investigated the association of some SNPs with resistance to salmonellosis. The Nramp1 cDNA was 1746 bp long and the Nramp2 cDNA was 1938 bp long. These cDNAs are similar to previously reported cDNAs, varying by two and one amino acids, respectively. The chicken Nramp1 gene expressed predominantly in liver, thymus and spleen in both females and males. The Nramp2 gene expressed in almost all tissues, but predominantly in breast muscle, leg muscle, cerebrum, cerebellum, lung, kidney, and heart in both females and males. We identified 45 SNPs and 2 indels in the chicken Nramp1 gene; three of 13 SNPs in the exons were missense mutations (Arg223Gln, Ala273Glu and Arg497Gln). Association analysis indicated that A24101991G is significantly associated with chicken salmonellosis resistance. These results will be useful for functional investigation of chicken Nramp1 and Nramp2 genes.


Is urban-rural location associated with weight status in school children? An examination of 42 small and rural Californian counties.
Studies have identified geographic variation in overweight and obesity rates among children, with higher rates of overweight and obesity often found among children living in rural compared to urban areas. A small number of studies have explored differences in overweight and obesity based on more nuanced gradations along the urban-rural continuum. The purpose of the present study was to identify differences in overweight and obesity based on gradations along the urban-rural continuum among children in 42 Californian counties with populations less than 500 000. An observational study was conducted using FITNESSGRAM data collected from 5th, 7th and 9th grade students in public schools in California during 2010-2011. The FITNESSGRAM dataset was merged with the 2011 Public Elementary/Secondary School Universe Survey Data from the National Center for Educational Statistics Common Core of Data, which includes an 'urban-centric locale' code for each school, consisting of four broad classifications - city, suburb, town, and rural - each of which is further broken down into three subcategories. Multivariate analyses using a general linear model were conducted to compare differences in body mass index (BMI) between geographic regions of schools (city, suburb, town and rural) as well as 11 urban-centric locale code subcategories; none of the schools were located in large cities. The percentage of students who were overweight and/or obese was compared by grade level, gender, and race/ethnicity across geographic regions using multivariate logistic regression models. Analyses were adjusted for student age, grade, gender, race/ethnicity (African-American, Asian, Hispanic, Indian/Alaskan, White, two or more races or unknown), eligibility for free or reduced price meals, and clustering of students by school. When a stratified analysis was done, the variable of stratification (ie grade, gender, race/ethnicity) was not included among the covariates. When significant differences in BMI or prevalence of overweight or obesity were found between geographic regions, Tukey's method was applied to adjust for multiple comparisons at a 5% procedure-wise error rate. A p-value at or less than 0.05 was used to indicate statistical significance. Students in suburban schools had significantly lower mean BMI and lower prevalence of overweight than students in other geographic areas (p<0.0001). Among 5th and 7th grade students, prevalence of obesity (but not overweight) varied by urban-rural status (p<0.0001, p=0.01, respectively), with 7th grade students in suburbs having lower rates of obesity than those in towns. Among 9th grade students, prevalence of overweight (but not obesity) varied by urban-rural status (p=0.02). Among females, prevalence of overweight and obesity varied (p=0.006, p<0.0001, respectively), with suburbs having lower rates than cities and towns. Among males, prevalence of obesity varied (p<0.0001), with suburbs having lower rates. Among whites, there were differences in prevalence of overweight and obesity by urban-rural status (p=0.01, p <0.0001, respectively). Among Hispanics, the prevalence of obesity varied by urban-rural status (p=0.001). Large suburban areas had the lowest rates of obesity compared to all other subcategories. Students attending schools in suburban, especially larger suburban, areas appear to have lower prevalence of obesity than their peers at schools in other geographic areas. Further research is needed to understand the factors associated with differences in weight status between urban, suburban, town and rural areas.


Ln[DO3A-N-α-(pyrenebutanamido)propionate] complexes: optimized relaxivity and NIR optical properties.
We have proposed recently that the DO3A-N-α-(amino)propionate chelator and its amide conjugates are leads to targeted, high relaxivity, safe contrast agents for magnetic resonance imaging. In this work we illustrate further the expeditious nature and robustness of the synthetic methodologies developed by preparing the DO3A-N-(α-pyrenebutanamido)propionate chelator. Its Gd(3+) chelate retains the optimized water exchange, high stability and inertness of the parent complex. The pyrene moiety imparts concentration-dependent self-assembly properties and aggregation-sensitive fluorescence emission to the Gd(3+) complex. The Gd(3+) complex displays pyrene-centred fluorescence whilst the Yb(3+) and Nd(3+) complexes exhibit sensitized lanthanide-centred near-infrared luminescence. The aggregated form of the complex displays high relaxivity (32 mM(-1) s(-1), 20 MHz, 25 °C) thanks to simultaneous optimization of the rotational correlation time and of the water exchange rate. The relaxivity is however still limited by chelate flexibility. This report demonstrates that the DO3A-N-(α-amino)propionate chelator is a valuable platform for constructing high relaxivity CA using simple design principles and robust chemistries accessible to most chemistry labs.


Intimate partner violence among economically disadvantaged young adult women: associations with adolescent risk-taking and pregnancy experiences.
Intimate partner violence negatively impacts the health of substantial proportions of young women in economically disadvantaged communities, where sexual initiation, aggressive behaviors, unintended pregnancies and childbearing are common among adolescents. It is therefore important to assess how adolescent risk behaviors and pregnancy experiences are linked to such violence during young adulthood. Data from 526 participants in the Reach for Health Longitudinal Study who were surveyed during middle school (in 1995-1996 and 1996-1997) and at ages 22-25 (in 2005-2007) provided information on adolescent risk behaviors and pregnancy experiences, as well as experiences of intimate partner violence during young adulthood. Bivariate and multivariate analyses were conducted to identify correlates of intimate partner violence involvement. As young adults, 29% of women reported having been victims of intimate partner violence in the past 12 months; 21% reported having perpetrated such violence. In multivariate analyses, victimization and perpetration in the last year are positively associated with aggressive behavior in middle school (odds ratios, 1.9 and 2.5, respectively), lifetime number of sex partners (1.3 for both) and having a history of unintended pregnancy or pregnancy problems (1.3 for both). Perpetration also is associated with early sexual initiation (0.5) and living with a partner (1.8). It is important to consider women's pregnancy histories in programs aimed at preventing the adverse outcomes of relationship violence and in screening for partner violence in sexual and reproductive health services. Early intervention may help women develop the skills needed for resolving conflicts with peers and partners.


Dermal injection of radioactive colloid is superior to peritumoral injection for breast cancer sentinel lymph node biopsy: results of a multiinstitutional study.
To determine the optimal radioactive colloid injection technique for sentinel lymph node (SLN) biopsy for breast cancer. The optimal radioactive colloid injection technique for breast cancer SLN biopsy has not yet been defined. Peritumoral injection of radioactive colloid has been used in most studies. Although dermal injection of radioactive colloid has been proposed, no published data exist to establish the false-negative rate associated with this technique. The University of Louisville Breast Cancer Sentinel Lymph Node Study is a multiinstitutional study involving 229 surgeons. Patients with clinical stage T1-2, N0 breast cancer were eligible for the study. All patients underwent SLN biopsy, followed by level I/II axillary dissection. Peritumoral, subdermal, or dermal injection of radioactive colloid was performed at the discretion of the operating surgeon. Peritumoral injection of isosulfan blue dye was performed concomitantly in most patients. The SLN identification rates and false-negative rates were compared. The ratios of the transcutaneous and ex vivo radioactive SLN count to the final background count were calculated as a measure of the relative degree of radioactivity of the nodes. One-way analysis of variance and chi-square tests were used for statistical analysis. A total of 2,206 patients were enrolled. Peritumoral, subdermal, or dermal injection of radioactive colloid was performed in 1,074, 297, and 511 patients, respectively. Most of the patients (94%) who underwent radioactive colloid injection also received peritumoral blue dye injection. The SLN identification rate was improved by the use of dermal injection compared with subdermal or peritumoral injection of radioactive colloid. The false-negative rates were 9.5%, 7.8%, and 6.5% (not significant) for peritumoral, subdermal, and dermal injection techniques, respectively. The relative degree of radioactivity of the SLN was five- to sevenfold higher with the dermal injection technique compared with peritumoral injection. Dermal injection of radioactive colloid significantly improves the SLN identification rate compared with peritumoral or subdermal injection. The false-negative rate is also minimized by the use of dermal injection. Dermal injection also is associated with SLNs that are five- to sevenfold more radioactive than with peritumoral injection, which simplifies SLN localization and may shorten the learning curve.


GLIF3: the evolution of a guideline representation format.
The Guideline Interchange Format (GLIF) is a language for structured representation of guidelines. It was developed to facilitate sharing clinical guidelines. GLIF version 2 enabled modeling a guideline as a flowchart of structured steps, representing clinical actions and decisions. However, the attributes of structured constructs were defined as text strings that could not be parsed, and such guidelines could not be used for computer-based execution that requires automatic inference. GLIF3 is a new version of GLIF designed to support computer-based execution. GLIF3 builds upon the framework set by GLIF2 but augments it by introducing several new constructs and extending GLIF2 constructs to allow a more formal definition of decision criteria, action specifications and patient data. GLIF3 enables guideline encoding at three levels: a conceptual flowchart, a computable specification that can be verified for logical consistency and completeness, and an implementable specification that can be incorporated into particular institutional information systems.


Cautioning the use of degree-day models for climate change projections in the presence of parametric uncertainty.
Developmental models, such as degree-day models, are commonly used to predict the impact of future climate change on the intensity, distribution, and timing of the transmission of infectious diseases, particularly those caused by pathogens carried by vectors or intermediate hosts. Resulting projections can be useful in policy discussions concerning regional or national responses to future distributions of important infectious diseases. Although the simplicity of degree-day models is appealing, little work has been done to analyze their ability to make reliable projections of the distribution of important pathogens, vectors, or intermediate hosts in the presence of the often considerable parametric uncertainty common to such models. Here, a population model of Oncomelania hupensis, the intermediate host of Schistosoma japonicum, was used to investigate the sensitivity of host range predictions in Sichuan Province, China, to uncertainty in two key degree-day model parameters: delta(min) (minimum temperature threshold for development) and K (total degree-days required for completion of snail development). The intent was to examine the consequences of parametric uncertainty in a plausible biological model, rather than to generate the definitive model. Results indicate that model output, the seasonality of population dynamics, and range predictions, particularly along the edge of the range, are highly sensitive to changes in model parameters, even at levels of parametric uncertainty common to such applications. Caution should be used when interpreting the results of degree-day models used to generate predictions of disease distribution and risk under scenarios of future climate change, and predictions should be considered most reliable when the temperature ranges used in projections resemble those used to estimate model parameters. Given the potential for substantial changes in degree-day model output with modest changes in parameter values, caution is warranted when results will be used to inform policy and management decisions.


SARS-CoV-2 infection in the Italian Veneto region: adverse outcomes in patients with cancer.
In 84,246 consecutive Italians tested for SARS-CoV-2, we examined the prevalence of cancer and clinical outcomes of viral infection. Overall, 5.7% (4,789/84,246) had previously been diagnosed with cancer and, among the SARS-CoV-2-positive (CV2+ve) patients, 7.8% had a cancer diagnosis (723/9,275). Prevalence of cancer was not associated with risk of infection (odds ratio of 0.97; 95% confidence interval of 0.89-1.06). The proportion of CV2+ve people hospitalized was higher among patients with cancer (56.6 versus 34.4% among other people), and so was the proportion of deaths (14.7% versus 4.5%). The risk of adverse outcomes of SARS-CoV-2 infection was significantly higher for patients with cancer versus those without, particularly for males and older people.


The Cancer Stem Cell Fraction in Hierarchically Organized Tumors Can Be Estimated Using Mathematical Modeling and Patient-Specific Treatment Trajectories.
Many tumors are hierarchically organized and driven by a subpopulation of tumor-initiating cells (TIC), or cancer stem cells. TICs are uniquely capable of recapitulating the tumor and are thought to be highly resistant to radio- and chemotherapy. Macroscopic patterns of tumor expansion before treatment and tumor regression during treatment are tied to the dynamics of TICs. Until now, the quantitative information about the fraction of TICs from macroscopic tumor burden trajectories could not be inferred. In this study, we generated a quantitative method based on a mathematical model that describes hierarchically organized tumor dynamics and patient-derived tumor burden information. The method identifies two characteristic equilibrium TIC regimes during expansion and regression. We show that tumor expansion and regression curves can be leveraged to infer estimates of the TIC fraction in individual patients at detection and after continued therapy. Furthermore, our method is parameter-free; it solely requires the knowledge of a patient's tumor burden over multiple time points to reveal microscopic properties of the malignancy. We demonstrate proof of concept in the case of chronic myeloid leukemia (CML), wherein our model recapitulated the clinical history of the disease in two independent patient cohorts. On the basis of patient-specific treatment responses in CML, we predict that after one year of targeted treatment, the fraction of TICs increases 100-fold and continues to increase up to 1,000-fold after 5 years of treatment. Our novel framework may significantly influence the implementation of personalized treatment strategies and has the potential for rapid translation into the clinic. Cancer Res; 76(7); 1705-13. ©2016 AACR.


Governance Through Privacy, Fairness, and Respect for Individuals.
Individuals have a moral claim to be involved in the governance of their personal data. Individuals' rights include privacy, autonomy, and the ability to choose for themselves how they want to manage risk, consistent with their own personal values and life situations. The Fair Information Practices principles (FIPPs) offer a framework for governance. Privacy-enhancing technology that complies with applicable law and FIPPs offers a dynamic governance tool for enabling the fair and open use of individual's personal data. Any governance model must protect against the risks posed by data misuse. Individual perceptions of risks are a subjective function involving individuals' values toward self, family, and society, their perceptions of trust, and their cognitive decision-making skills. Individual privacy protections and individuals' right to choose are codified in the HIPAA Privacy Rule, which attempts to strike a balance between the dual goals of information flow and privacy protection. The choices most commonly given individuals regarding the use of their health information are binary ("yes" or "no") and immutable. Recent federal recommendations and law recognize the need for granular, dynamic choices. Individuals expect that they will govern the use of their own health and genomic data. Failure to build and maintain individuals' trust increases the likelihood that they will refuse to grant permission to access or use their data. The "no surprises principle" asserts that an individual's personal information should never be collected, used, transmitted, or disclosed in a way that would surprise the individual were she to learn about it. The FIPPs provide a powerful framework for enabling data sharing and use, while maintaining trust. We introduce the eight FIPPs adopted by the Department of Health and Human Services, and provide examples of their interpretation and implementation. Privacy risk and health risk can be reduced by giving consumers control, autonomy, and transparency, and by engaging them in managing their own health. Explicit "consent" may not always be necessary - the FIPPs offer multiple ways to engender trust and avoid surprises.


Individual antigen-specific T lymphocytes: helper function in enabling the expression of multiple antibody isotypes.
In recent years antigen-specific T cells have been shown to be capable of mediating a number of diverse functions in collaboration with B cells in humoral immune responses. One of the more intriguing roles attributed to helper T cells is the promotion of the synthesis of multiple immunoglobulin isotypes by B cells in T-dependent antibody responses. The experiments presented in this report were carried out to determine if an individual antigen-specific T lymphocyte has the capability to enable the production of antibodies of multiple immunoglobulin heavy chain isotypes. We describe an experimental system which allows for the isolation and antigenic stimulation of individual helper T cells in a splenic environment which provides an excess of primary B cells for collaboration with isolated T lymphocytes. Employing this system we have demonstrated that an individual antigen-specific T lymphocyte, specific for the PR8 strain of influenza virus, has the capacity to enable primary B-cell PR8-specific antibody responses of more than a single immunoglobulin isotype. The implications made by these studies regarding the problem of genetic restrictions regulating T-cell-B-cell interaction is discussed.


Extrinsic labeling method may not accurately measure Fe absorption from cooked pinto beans (Phaseolus vulgaris): comparison of extrinsic and intrinsic labeling of beans.
Isotopic labeling of food has been widely used for the measurement of Fe absorption in determining requirements and evaluating the factors involved in Fe bioavailability. An extrinsic labeling technique will not accurately predict the total Fe absorption from foods unless complete isotopic exchange takes place between an extrinsically added isotope label and the intrinsic Fe of the food. We examined isotopic exchange in the case of both white beans and colored beans (Phaseolus vulgaris) with an in vitro digestion model. There are significant differences in (58)Fe/(56)Fe ratios between the sample digest supernatant and the pellet of extrinsically labeled pinto bean. The white bean digest shows significantly better equilibration of the extrinsic (58)Fe with the intrinsic (56)Fe. In contrast to the extrinsically labeled samples, both white and red beans labeled intrinsically with (58)Fe demonstrated consistent ratios of (58)Fe/(56)Fe in the bean meal, digest, supernatant, and pellet. It is possible that the polyphenolics in the bean seed coat may bind Fe and thus interfere with extrinsic labeling of the bean meals. These observations raise questions on the accuracy of studies that used extrinsic tags to measure Fe absorption from beans. Intrinsic labeling appears necessary to accurately measure Fe bioavailability from beans.